PI3K/Akt/NF-κB介导的FSH对上皮性卵巢癌细胞增殖与侵袭机制的研究
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摘要
目的:卵巢恶性肿瘤是女性生殖器官三大恶性肿瘤之一,其早期诊断率低,中晚期转移率高,预后较差。对于卵巢癌发病机理的研究仍有许多问题有待于解决。近年来越来越多的证据表明,卵巢癌的发生和进展与其所处的激素环境有关。绝经,排卵多或不孕治疗时过量卵泡刺激素(FSH)刺激可能是卵巢癌发生的重要危险因素。多项研究显示FSH作用于细胞表面FSH受体激活胞内信号传导通路发挥作用,而其具体作用机制尚不清楚。磷脂酞肌醇3激酶/蛋白激酶B(PI3K/AKT)信号传导通路在肿瘤的发生、发展、凋亡、血管生成、转移及耐药等方面起着重要的作用。可被PI3K/AKT通路直接或间接影响的下游分子主要包括细胞增殖和蛋白合成的相关因子和一些凋亡相关因子。核转录因子(NF-κB)作为信号传导通路的中枢,是AKT重要的下游分子之一,其异常活化可调节与细胞增殖和凋亡有关的基因转录,溶解细胞外间质促进肿瘤细胞的浸润和转移。因此,本实验主要研究FSH通过PI3K/AKT途径作用于NF-κB,并调节其表达,使卵巢上皮性癌细胞发生增殖和侵袭的机制,以及阻断该通路对卵巢癌恶性进展的影响,拟为卵巢癌的发病机制和基因治疗提供新的实验依据。
方法:将SKOV-3、3AO细胞置于37℃,饱和湿度5%CO2的RPMI-1640培养基中常规培养,待细胞进入对数生长期后用于实验。
1光镜观察:将不同浓度的FSH和PI3K/AKT抑制剂LY294002作用于SKOV-3、3AO细胞,在倒置相差显微镜下定时观察细胞的生长情况并照相。
2 MTT法:调整细胞浓度为1×105/ml,以每孔100μl培养于96孔培养板中,待细胞进入对数生长期后,用不同浓度FSH(10、20、40、80、160mIU/ml)作用不同时间(12h、24h、48h、72h),采用四甲基偶氮唑蓝(MTT)法检测药物对细胞的生长促进作用及分析其与时间和浓度之间的关系。
3 Western blot蛋白印迹法:观察FSH和LY294002分别及序贯联合作用于SKOV-3、3AO细胞前后FSHR、AKT1/2、p-AKT、NF-kappaB蛋白的表达,以及细胞核与细胞浆中NF-κB蛋白的表达差异。
4 Transwell小室侵袭实验:调整细胞浓度为1×105/ml,每孔上层小室中加200μl细胞悬液,24孔板槽中加入500μl含10%胎牛血清的培养基。待细胞贴壁后,用FSH 40mIU/ml和LY294002 10μmol/L分别及序贯联合作用于细胞,48小时后结晶紫染色,显微镜下观察上层小室底部穿过的细胞情况并计数。
5应用SPSS13.0统计软件进行统计学分析。
结果:
1倒置相差显微镜下观察可见经FSH作用SKOV-3、3AO细胞后,随时间延长,浓度增加,细胞形态稍变长,生长速度和密集度明显增加。经LY294002作用30分钟后加入FSH继续作用,细胞形态变化不大,增长速度和密集度减低。单独LY294002作用后部分细胞形状由梭形变为圆形,然后细胞开始皱缩,细胞增长能力减弱。
2不同浓度FSH(10、20、40、80、160mIU/ml)作用不同时间(12h、24h、48h、72h),MTT结果显示SKOV-3细胞于FSH 40mIU/ml作用48 h时细胞增长最快,浓度与其他组比较有显著差异(P<0.05),而时间点之间相比则无差异(P>0.05)。3AO于FSH 40mIU/ml作用24h时细胞增长最快,与其他浓度和时间点进行统计学比较均有显著差异(P<0.05)。
3 Western bloting结果:①SKOV-3、3AO细胞中FSHR蛋白均有表达,FSH和LY294002单独及联合处理后细胞表达与对照组相比无差异(P>0.05)。②SKOV-3细胞经FSH及LY294002单独处理与对照组相比分别表现为前者p-AKT蛋白表达增加,后者表达则减少(P<0.05),FSH/LY294002联合处理与对照组比较表达减少(P<0.05),而AKT1/2蛋白表达四组间均无差异(P>0.05)。NF-κB总蛋白在三个实验组中表达与对照组相比均无统计学意义(P>0.05);但FSH组表达明显多于LY294002组(P<0.05)。胞核中NF-κB蛋白表达FSH组与对照组相比表达增加(P<0.05),而LY294002组及FSH/LY294002组表达减少(P>0.05);且FSH组表达明显多于LY294002组、FSH/LY294002组(P<0.05),而后两者比较无差异(P>0.05)。胞浆中NF-κB蛋白三个实验组与对照组相比表达均增加,但无统计学差异(P>0.05)。对对照组、FSH组、FSH/LY294002组及LY294002组各组中胞核与胞浆的比值进行统计学分析,四组间有显著差异(P<0.05)。其中,FSH组明显大于FSH/LY294002组及LY294002组,LY294002组明显小于对照组和FSH组(P<0.05),而其他组间比较无差异(P>0.05)。③3AO细胞中p-AKT蛋白表达,三个实验组与对照组比较均有统计学差异(P<0.05)。其中,FSH组p-AKT蛋白表达明显多于其余三组(P<0.05);LY294002组、FSH/LY294002组中表达明显少于对照组(P<0.05)。四组之间AKT1/2蛋白表达均无差异(P>0.05)。总提取蛋白中NF-κB蛋白表达FSH组明显少于对照组及LY294002组(P<0.05),而四组之间及其他组间比较无明显差异(P>0.05)。胞核中NF-κB蛋白表达四组之间比较有明显差异(P<0.05),其中,FSH组与其他三组相比表达明显增加(P<0.05);而LY294002组,FSH/LY294002组与对照组比较表达减少,但无统计学差异(P>0.05),且其两组间比较亦无差异(P>0.05);胞浆中NF-κB蛋白表达FSH组多于对照组及LY294002组、FSH/LY294002组,但组组间比较无统计学差异(P>0.05)。对对照组、FSH组、FSH/LY294002组及LY294002组各组中胞核与胞浆的比值进行统计学分析,四组间有显著差异(P<0.05)。其中,FSH组明显大于其余三组(P<0.05),FSH/LY294002组明显大于对照组(P<0.05),而FSH/LY294002组与LY294002组比较无差异(P>0.05)。
4 Transwell小室侵袭结果:经FSH处理的两种细胞穿过小室的细胞数明显多于对照组(P<0.05);经LY294002单独处理组及两者联合作用组的两种细胞穿过小室的细胞数均明显少于FSH组(P<0.05),但两组相比无明显差异(P>0.05)。
结论:
1卵泡刺激素有促进上皮性卵巢癌细胞株SKOV-3、3AO增殖作用,提示增殖峰最高时的浓度可能是最强作用浓度。
2 SKOV-3、3AO细胞中均存在FSHR受体,卵泡刺激素作用于细胞SKOV-3、3AO后激活PI3K/Akt途径,提高了p-AKT蛋白的表达量,进而诱导其下游分子NF-κB蛋白在细胞核中表达增加,促进细胞增殖,增加肿瘤细胞侵袭能力。
3 PI3K/Akt抑制剂LY294002作用于SKOV-3、3AO细胞后再给予FSH刺激,p-AKT活性被抑制,蛋白表达明显降低,进而NF-κB蛋白在细胞核中表达降低,抑制细胞增殖,降低肿瘤细胞侵袭能力。
4卵泡刺激素可能是通过PI3K/Akt通路促进卵巢癌细胞株SKOV-3、3AO中NF-κB的表达,实现对卵巢癌细胞的促增殖和侵袭作用。
Objective: Ovarian cancer is one of the three female genitalia malignant tumor, and has low early diagnosis, high metastasis rate and poor prognosis. So the pathogenesis of ovarian cancer still has many problems to be solved. There is increasing evidences suggesting that it is the hormonal environment of the normal ovarian surface epithelium (OSE) and ovarian epithelial cancer (OEC) cells that is associated with the development and progression of ovarian cancer. Exposure to excess follicle stimulating hormone (FSH), related to menopause, ovulation, or infertility therapy, has been implicated as an important possible risk factor for ovarian cancer. A number of studies have shown that FSH acting on FSH receptor in the cell surface play a role by activating of intracellular signal transduction pathways, but its specific mechanism of action is unknown. Phosphatidylinositols-kinase/protein kinase B (PI3K/AKT) signaling pathway play an important role in tumor occurrence, development, apoptosis, angiogenesis, metastasis and drug resistance, etc. PI3K/AKT pathway may be directly or indirectly affect the downstream molecules include cell proliferation and protein synthesis related factors and a number of apoptosis related factors. Nuclear transcription factor (NF-κB) as a center of signal transduction pathway, is one of important downstream molecule of AKT. Its abnormal activation may regulate cell proliferation and apoptosis related gene transcription, dissolved in extracellular matrix to promote tumor cell infiltration and metastasis. Therefore, this experiment mainly study the role of FSH on NF-κB through PI3K/AKT pathway and to regulate its expression, get the occurrence mechanism of epithelial ovarian cancer cell proliferation and invasion, and the malignant progression of ovarian cancer by blocking the pathway. This study will provide new experimental evidence for the pathogenesis of ovarian cancer and gene therapy.
Methods: SKOV-3, 3AO cells were cultured at 37℃in RPMI-1640 in the tissue culture flask under a humidified 5%CO2 atmosphere, applying to the experiment when they entered the logarithmic phase .
1 Light microscopy observation of cell morphological changes: different concentrations of FSH and PI3K/AKT inhibitor LY294002 were roled in SKOV-3, 3AO cells and to observe the growth of cell regularly in inverted phase contrast microscope and to photo.
2 MTT method: adjusting the cell concentration of 1×105/ml, with 100μl per well cultured in 96-well plate. Then the cells were exposed in different concentrations of FSH (10, 20, 40, 80, 160 mIU/ml) when the cells entered logarithmic growth phase. At last MTT method detected the proliferation of cell growth in different times(12h, 24h, 48h, 72h).
3 Western blot analyses: To observe FSHR, Akt1 / 2, p-AKT, NF-kappaB protein expression of SKOV-3, 3AO cells before and after exposed to FSH and LY294002 respectively or combined by western blot, as well as the different expression of NF-kappaB protein in the nucleus and cytoplasm.
4 Transwell invasion method: adjusting the cell concentration of 1×105/ml, with 200μl cell suspension in the inner cup of the 24-well Corning chamber that had been coated with 40μl Matrigel and 500μl RMPI-1640 culture media supplemented with 10% heat-inactivated FBS in the outer cup. Then the cells were exposed in FSH 40mIU/ml and LY294002 10μmol/L respectively or combined until adherencing. After 48 h, cells that had invaded through the Matrigel were fixed, stained, then observed and counted under the light microscope.
5 Application of SPSS13.0 statistical software for statistical analysis.
Results:
1 The appearance of SKOV-3, 3AO cells exposed to FSH were observed under inverted phase contrast microscope. As the concentration increased and the time extended, the shape of the cells began to change slightly longer, the growth rate and intensity increased significantly. After LY294002 treating 30 minutes, FSH was exposed to continue to effect, leading to little change in cell morphology, growth rate and intensity reduced. Exposing LY294002 alone, some cells were from the spindle into a round shape, and then began to shrink. The cell growth capacity diminished.
2 After we treated SKOV-3, 3AO cells in different time (12h, 24h, 48h, and 72h ) and with FSH in different concentration(s10,20,40,80,160 mIU/ml), MTT results showed that SKOV-3 cells were in the fastest growing for 48 h and FSH in 40μmol/ml, the concentration compared with the other groups was significantly different (P<0.05), and between the time groups there was no differences (P>0.05). 3AO cells were for 24 h and in 40μmol/ml, and compared with the other concentrations and times has significant differences (p <0.05).
3 Result of western blotting:①FSHR protein were expressed in SKOV-3, 3AO cell. After FSH and LY294002 alone or in combination treating the cells, there were no significant differences as compared with the control group (p> 0.05).②A fter treating SKOV-3 cells with FSH and LY294002 alone, compared with the control group showed that p-AKT protein expression increased in FSH group, and the expression decreased in LY294002 group (p <0.05). In FSH/LY294002 combined treatement, the expression decreased compared with the control group (P<0.05). And AKT1 / 2 protein expression has no significant differences among the four groups (P>0.05). NF-κB total protein expression in the three experimental groups as compared with the control group, there was no statistical significance (P>0.05); but FSH group has more expression than LY294002 group (P<0.05). NF-κB protein expression in the nucleus of FSH group, compared with the control group increased (P<0.05), while the expression of LY294002 and FSH/LY294002 group decreased (P>0.05); and the expression of FSH group more than LY294002 group, FSH /LY294002 group (P<0.05). The latter two showed no difference (P>0.05). NF-κB protein expression in the cytoplasm of three experimental groups as compared with the control group had increased (p> 0.05). Analyzed statistically the ratio of the nucleus and the cytoplasm in the control group, FSH group, FSH/LY294002 group and LY294002 group, the four groups were significantly different (P<0.05). Which, FSH significantly larger than FSH/LY294002 group and LY294002 group, LY294002 group was significantly less than the control group and the FSH group (P<0.05), while there were no differences between the other groups (P>0.05).③p-AKT protein expression in 3AO cells, the three experimental groups compared with control group were statistically different (P<0.05). Which, p-AKT protein expression of FSH group was significantly more than the other three groups (p <0.05); LY294002 group, FSH/LY294002 group expressed significantly lower than the control group (P<0.05). Among the four groups AKT1/2 protein expression had on difference (P>0.05). NF-κB protein expression of total extracted protein, FSH group was significantly less than the control group and LY294002 group (P<0.05), while the other groups and between the four groups showed no significant difference (P>0.05). NF-κB protein expression of the nucleus in the four groups have more significant differences (p <0.05), which, the expression of FSH group increased significantly compared with the other three groups (P<0.05); while the expression of LY294002 group, FSH/LY294002 group decreased compared with the control group, but no statistically significant difference (P>0.05), and no difference between the two groups (P>0.05); NF-κB protein expression of the cytoplasm in FSH group were more than the control group and LY294002 group, FSH/LY294002 group, but between the latter two groups was no significant difference (P>0.05). Analyzed statistically the ratio of the nucleus and the cytoplasm in the control group, FSH group, FSH/LY294002 group and LY294002 group, the four groups were significantly different (P<0.05). Which, FSH group was significantly higher than the other three groups (P<0.05), FSH/LY294002 group was significantly more than the control group (P<0.05), while the FSH/LY294002 group and LY294002 group had no significant difference (p> 0.05).
4 The results of Transwell invasion method: Treating the two kinds of cells with FSH, the number of cells invading through the Matrigel were more than the control group (P<0.05). LY294002 treatment alone group and combined of the two drugs group also had a few cells invading through. And the cells were significantly less than FSH group (P<0.05), but compared the two groups had no significant differences (P>0.05).
Conclusion:
1 Follicle-stimulating hormone has the effect of inducing proliferation on epithelial ovarian cancer cell line SKOV-3, 3AO. That suggests that the peak concentration may be the strongest proliferative concentration.
2 FSH receptor exist in both of SKOV-3, 3AO cells. That FSH acts on SKOV-3, 3AO cells could excite PI3K/Akt pathway. So p-AKT protein expression increase. Thereby, inducing its downstream molecule NF -κB protein expression increase in the nucleus, promoted cell proliferation and invasion ability.
3 PI3K/Akt inhibitor LY294002 acts on SKOV-3, 3AO cells and then given to FSH stimulation, p-AKT activity is inhibited, protein expression is significantly reduced, and thus NF-κB protein expression in the nucleus is reduced, inhibiting cell proliferation, reducing the invasive ability of tumor cells.
4 Follicle-stimulating hormone may be through PI3K/Akt pathway to promote ovarian cancer cell line SKOV-3, 3AO in the expression of NF-κB, then achieve on promoting the proliferation and invasion of ovarian cancer cells.
方法:将SKOV-3、3AO细胞置于37℃,饱和湿度5%CO2的RPMI-1640培养基中常规培养,待细胞进入对数生长期后用于实验。
1光镜观察:将不同浓度的FSH和PI3K/AKT抑制剂LY294002作用于SKOV-3、3AO细胞,在倒置相差显微镜下定时观察细胞的生长情况并照相。
2 MTT法:调整细胞浓度为1×105/ml,以每孔100μl培养于96孔培养板中,待细胞进入对数生长期后,用不同浓度FSH(10、20、40、80、160mIU/ml)作用不同时间(12h、24h、48h、72h),采用四甲基偶氮唑蓝(MTT)法检测药物对细胞的生长促进作用及分析其与时间和浓度之间的关系。
3 Western blot蛋白印迹法:观察FSH和LY294002分别及序贯联合作用于SKOV-3、3AO细胞前后FSHR、AKT1/2、p-AKT、NF-kappaB蛋白的表达,以及细胞核与细胞浆中NF-κB蛋白的表达差异。
4 Transwell小室侵袭实验:调整细胞浓度为1×105/ml,每孔上层小室中加200μl细胞悬液,24孔板槽中加入500μl含10%胎牛血清的培养基。待细胞贴壁后,用FSH 40mIU/ml和LY294002 10μmol/L分别及序贯联合作用于细胞,48小时后结晶紫染色,显微镜下观察上层小室底部穿过的细胞情况并计数。
5应用SPSS13.0统计软件进行统计学分析。
结果:
1倒置相差显微镜下观察可见经FSH作用SKOV-3、3AO细胞后,随时间延长,浓度增加,细胞形态稍变长,生长速度和密集度明显增加。经LY294002作用30分钟后加入FSH继续作用,细胞形态变化不大,增长速度和密集度减低。单独LY294002作用后部分细胞形状由梭形变为圆形,然后细胞开始皱缩,细胞增长能力减弱。
2不同浓度FSH(10、20、40、80、160mIU/ml)作用不同时间(12h、24h、48h、72h),MTT结果显示SKOV-3细胞于FSH 40mIU/ml作用48 h时细胞增长最快,浓度与其他组比较有显著差异(P<0.05),而时间点之间相比则无差异(P>0.05)。3AO于FSH 40mIU/ml作用24h时细胞增长最快,与其他浓度和时间点进行统计学比较均有显著差异(P<0.05)。
3 Western bloting结果:①SKOV-3、3AO细胞中FSHR蛋白均有表达,FSH和LY294002单独及联合处理后细胞表达与对照组相比无差异(P>0.05)。②SKOV-3细胞经FSH及LY294002单独处理与对照组相比分别表现为前者p-AKT蛋白表达增加,后者表达则减少(P<0.05),FSH/LY294002联合处理与对照组比较表达减少(P<0.05),而AKT1/2蛋白表达四组间均无差异(P>0.05)。NF-κB总蛋白在三个实验组中表达与对照组相比均无统计学意义(P>0.05);但FSH组表达明显多于LY294002组(P<0.05)。胞核中NF-κB蛋白表达FSH组与对照组相比表达增加(P<0.05),而LY294002组及FSH/LY294002组表达减少(P>0.05);且FSH组表达明显多于LY294002组、FSH/LY294002组(P<0.05),而后两者比较无差异(P>0.05)。胞浆中NF-κB蛋白三个实验组与对照组相比表达均增加,但无统计学差异(P>0.05)。对对照组、FSH组、FSH/LY294002组及LY294002组各组中胞核与胞浆的比值进行统计学分析,四组间有显著差异(P<0.05)。其中,FSH组明显大于FSH/LY294002组及LY294002组,LY294002组明显小于对照组和FSH组(P<0.05),而其他组间比较无差异(P>0.05)。③3AO细胞中p-AKT蛋白表达,三个实验组与对照组比较均有统计学差异(P<0.05)。其中,FSH组p-AKT蛋白表达明显多于其余三组(P<0.05);LY294002组、FSH/LY294002组中表达明显少于对照组(P<0.05)。四组之间AKT1/2蛋白表达均无差异(P>0.05)。总提取蛋白中NF-κB蛋白表达FSH组明显少于对照组及LY294002组(P<0.05),而四组之间及其他组间比较无明显差异(P>0.05)。胞核中NF-κB蛋白表达四组之间比较有明显差异(P<0.05),其中,FSH组与其他三组相比表达明显增加(P<0.05);而LY294002组,FSH/LY294002组与对照组比较表达减少,但无统计学差异(P>0.05),且其两组间比较亦无差异(P>0.05);胞浆中NF-κB蛋白表达FSH组多于对照组及LY294002组、FSH/LY294002组,但组组间比较无统计学差异(P>0.05)。对对照组、FSH组、FSH/LY294002组及LY294002组各组中胞核与胞浆的比值进行统计学分析,四组间有显著差异(P<0.05)。其中,FSH组明显大于其余三组(P<0.05),FSH/LY294002组明显大于对照组(P<0.05),而FSH/LY294002组与LY294002组比较无差异(P>0.05)。
4 Transwell小室侵袭结果:经FSH处理的两种细胞穿过小室的细胞数明显多于对照组(P<0.05);经LY294002单独处理组及两者联合作用组的两种细胞穿过小室的细胞数均明显少于FSH组(P<0.05),但两组相比无明显差异(P>0.05)。
结论:
1卵泡刺激素有促进上皮性卵巢癌细胞株SKOV-3、3AO增殖作用,提示增殖峰最高时的浓度可能是最强作用浓度。
2 SKOV-3、3AO细胞中均存在FSHR受体,卵泡刺激素作用于细胞SKOV-3、3AO后激活PI3K/Akt途径,提高了p-AKT蛋白的表达量,进而诱导其下游分子NF-κB蛋白在细胞核中表达增加,促进细胞增殖,增加肿瘤细胞侵袭能力。
3 PI3K/Akt抑制剂LY294002作用于SKOV-3、3AO细胞后再给予FSH刺激,p-AKT活性被抑制,蛋白表达明显降低,进而NF-κB蛋白在细胞核中表达降低,抑制细胞增殖,降低肿瘤细胞侵袭能力。
4卵泡刺激素可能是通过PI3K/Akt通路促进卵巢癌细胞株SKOV-3、3AO中NF-κB的表达,实现对卵巢癌细胞的促增殖和侵袭作用。
Objective: Ovarian cancer is one of the three female genitalia malignant tumor, and has low early diagnosis, high metastasis rate and poor prognosis. So the pathogenesis of ovarian cancer still has many problems to be solved. There is increasing evidences suggesting that it is the hormonal environment of the normal ovarian surface epithelium (OSE) and ovarian epithelial cancer (OEC) cells that is associated with the development and progression of ovarian cancer. Exposure to excess follicle stimulating hormone (FSH), related to menopause, ovulation, or infertility therapy, has been implicated as an important possible risk factor for ovarian cancer. A number of studies have shown that FSH acting on FSH receptor in the cell surface play a role by activating of intracellular signal transduction pathways, but its specific mechanism of action is unknown. Phosphatidylinositols-kinase/protein kinase B (PI3K/AKT) signaling pathway play an important role in tumor occurrence, development, apoptosis, angiogenesis, metastasis and drug resistance, etc. PI3K/AKT pathway may be directly or indirectly affect the downstream molecules include cell proliferation and protein synthesis related factors and a number of apoptosis related factors. Nuclear transcription factor (NF-κB) as a center of signal transduction pathway, is one of important downstream molecule of AKT. Its abnormal activation may regulate cell proliferation and apoptosis related gene transcription, dissolved in extracellular matrix to promote tumor cell infiltration and metastasis. Therefore, this experiment mainly study the role of FSH on NF-κB through PI3K/AKT pathway and to regulate its expression, get the occurrence mechanism of epithelial ovarian cancer cell proliferation and invasion, and the malignant progression of ovarian cancer by blocking the pathway. This study will provide new experimental evidence for the pathogenesis of ovarian cancer and gene therapy.
Methods: SKOV-3, 3AO cells were cultured at 37℃in RPMI-1640 in the tissue culture flask under a humidified 5%CO2 atmosphere, applying to the experiment when they entered the logarithmic phase .
1 Light microscopy observation of cell morphological changes: different concentrations of FSH and PI3K/AKT inhibitor LY294002 were roled in SKOV-3, 3AO cells and to observe the growth of cell regularly in inverted phase contrast microscope and to photo.
2 MTT method: adjusting the cell concentration of 1×105/ml, with 100μl per well cultured in 96-well plate. Then the cells were exposed in different concentrations of FSH (10, 20, 40, 80, 160 mIU/ml) when the cells entered logarithmic growth phase. At last MTT method detected the proliferation of cell growth in different times(12h, 24h, 48h, 72h).
3 Western blot analyses: To observe FSHR, Akt1 / 2, p-AKT, NF-kappaB protein expression of SKOV-3, 3AO cells before and after exposed to FSH and LY294002 respectively or combined by western blot, as well as the different expression of NF-kappaB protein in the nucleus and cytoplasm.
4 Transwell invasion method: adjusting the cell concentration of 1×105/ml, with 200μl cell suspension in the inner cup of the 24-well Corning chamber that had been coated with 40μl Matrigel and 500μl RMPI-1640 culture media supplemented with 10% heat-inactivated FBS in the outer cup. Then the cells were exposed in FSH 40mIU/ml and LY294002 10μmol/L respectively or combined until adherencing. After 48 h, cells that had invaded through the Matrigel were fixed, stained, then observed and counted under the light microscope.
5 Application of SPSS13.0 statistical software for statistical analysis.
Results:
1 The appearance of SKOV-3, 3AO cells exposed to FSH were observed under inverted phase contrast microscope. As the concentration increased and the time extended, the shape of the cells began to change slightly longer, the growth rate and intensity increased significantly. After LY294002 treating 30 minutes, FSH was exposed to continue to effect, leading to little change in cell morphology, growth rate and intensity reduced. Exposing LY294002 alone, some cells were from the spindle into a round shape, and then began to shrink. The cell growth capacity diminished.
2 After we treated SKOV-3, 3AO cells in different time (12h, 24h, 48h, and 72h ) and with FSH in different concentration(s10,20,40,80,160 mIU/ml), MTT results showed that SKOV-3 cells were in the fastest growing for 48 h and FSH in 40μmol/ml, the concentration compared with the other groups was significantly different (P<0.05), and between the time groups there was no differences (P>0.05). 3AO cells were for 24 h and in 40μmol/ml, and compared with the other concentrations and times has significant differences (p <0.05).
3 Result of western blotting:①FSHR protein were expressed in SKOV-3, 3AO cell. After FSH and LY294002 alone or in combination treating the cells, there were no significant differences as compared with the control group (p> 0.05).②A fter treating SKOV-3 cells with FSH and LY294002 alone, compared with the control group showed that p-AKT protein expression increased in FSH group, and the expression decreased in LY294002 group (p <0.05). In FSH/LY294002 combined treatement, the expression decreased compared with the control group (P<0.05). And AKT1 / 2 protein expression has no significant differences among the four groups (P>0.05). NF-κB total protein expression in the three experimental groups as compared with the control group, there was no statistical significance (P>0.05); but FSH group has more expression than LY294002 group (P<0.05). NF-κB protein expression in the nucleus of FSH group, compared with the control group increased (P<0.05), while the expression of LY294002 and FSH/LY294002 group decreased (P>0.05); and the expression of FSH group more than LY294002 group, FSH /LY294002 group (P<0.05). The latter two showed no difference (P>0.05). NF-κB protein expression in the cytoplasm of three experimental groups as compared with the control group had increased (p> 0.05). Analyzed statistically the ratio of the nucleus and the cytoplasm in the control group, FSH group, FSH/LY294002 group and LY294002 group, the four groups were significantly different (P<0.05). Which, FSH significantly larger than FSH/LY294002 group and LY294002 group, LY294002 group was significantly less than the control group and the FSH group (P<0.05), while there were no differences between the other groups (P>0.05).③p-AKT protein expression in 3AO cells, the three experimental groups compared with control group were statistically different (P<0.05). Which, p-AKT protein expression of FSH group was significantly more than the other three groups (p <0.05); LY294002 group, FSH/LY294002 group expressed significantly lower than the control group (P<0.05). Among the four groups AKT1/2 protein expression had on difference (P>0.05). NF-κB protein expression of total extracted protein, FSH group was significantly less than the control group and LY294002 group (P<0.05), while the other groups and between the four groups showed no significant difference (P>0.05). NF-κB protein expression of the nucleus in the four groups have more significant differences (p <0.05), which, the expression of FSH group increased significantly compared with the other three groups (P<0.05); while the expression of LY294002 group, FSH/LY294002 group decreased compared with the control group, but no statistically significant difference (P>0.05), and no difference between the two groups (P>0.05); NF-κB protein expression of the cytoplasm in FSH group were more than the control group and LY294002 group, FSH/LY294002 group, but between the latter two groups was no significant difference (P>0.05). Analyzed statistically the ratio of the nucleus and the cytoplasm in the control group, FSH group, FSH/LY294002 group and LY294002 group, the four groups were significantly different (P<0.05). Which, FSH group was significantly higher than the other three groups (P<0.05), FSH/LY294002 group was significantly more than the control group (P<0.05), while the FSH/LY294002 group and LY294002 group had no significant difference (p> 0.05).
4 The results of Transwell invasion method: Treating the two kinds of cells with FSH, the number of cells invading through the Matrigel were more than the control group (P<0.05). LY294002 treatment alone group and combined of the two drugs group also had a few cells invading through. And the cells were significantly less than FSH group (P<0.05), but compared the two groups had no significant differences (P>0.05).
Conclusion:
1 Follicle-stimulating hormone has the effect of inducing proliferation on epithelial ovarian cancer cell line SKOV-3, 3AO. That suggests that the peak concentration may be the strongest proliferative concentration.
2 FSH receptor exist in both of SKOV-3, 3AO cells. That FSH acts on SKOV-3, 3AO cells could excite PI3K/Akt pathway. So p-AKT protein expression increase. Thereby, inducing its downstream molecule NF -κB protein expression increase in the nucleus, promoted cell proliferation and invasion ability.
3 PI3K/Akt inhibitor LY294002 acts on SKOV-3, 3AO cells and then given to FSH stimulation, p-AKT activity is inhibited, protein expression is significantly reduced, and thus NF-κB protein expression in the nucleus is reduced, inhibiting cell proliferation, reducing the invasive ability of tumor cells.
4 Follicle-stimulating hormone may be through PI3K/Akt pathway to promote ovarian cancer cell line SKOV-3, 3AO in the expression of NF-κB, then achieve on promoting the proliferation and invasion of ovarian cancer cells.
引文
1乐杰主编妇产科学[M].北京:人民卫生出版社.第6版.2004.305
2 Keith D, Edmonds, Dewhurst Text book of Obstetrics & Gynaecology. UK:Black well Publishing, seventh edition.2007.625
3 Christine H, Holschneicerm MD, Jonathan S, et al.Advances in the understanding and treatment of ovarian cancer. Brmeno-pause Soc, 2005, 11(2):66-71
4 Boso CK. Role of nerve growth factor and FSH receptor in epithelial ovarian cancer. Reprod Biomed Online,2005 11(2):194-197
5 Choi JH, Wong AS, Endocr Rev.Gonadotropins and Ovarian Cancer 2007 Jun; 28(4):440-61
6 Huang Y, Hua K.Activation of the PI3K/AKT pathway mediates FSH stimulated VEGF expression in ovarian serous cystadenocarcinoma. CellRes. 2008 Jul;18(7):780-91
7 Min C, Eddy SF. NF-kappaB and epithelial to mesenchymal transition of cancer. J Cell Biochem. 2008 Jun 1;104(3):733-44
8 Wang Y, Chan S, Tsang BK.Involvement of inhibitory nuclear factor kappaB activation in the gonadotropic regulation of X-linked inhibitor of apoptosis expression during ovarian follicular development in vitro. Endocrinology. 2002 Jul;143(7):2732-40
9 Bani MR, Nieoletti Ml, Aikarouf NW, et al.Gene expression correlating with response to paclitaxel in ovarian carcinoma xenografts. Mol Cancer Ther, 2004,3(2):111-121
10丰有吉,张惜阴,葛柏青,等.促性腺激素对人卵巢上皮性癌细胞的增殖作用.中华妇产科杂志,1996,31:166-8
11 Maruuehi T, Sugiyama T,Kataoka A,et al.Effects of a gonadotropin releasing hormone agonist on rat ovarian adenocarcinoma cell lines in vitro and in vivo. JPnJ Cancer Res,1998,89:977-83
12朱铭伟,吴乾渝,葛柏青等卵泡刺激素对卵巢癌细胞的增殖及CA125Ⅱ抗原分泌的作用.中华妇产科杂志,,1998,33:225-6
13 Syedy, UlinskiG, MokSC, et al.Expression of gonadotropin receptor and growth responses to key reproduetive hormones in normal and malignant human ovarian surface epithelial cells.Cancer Res, 2001, 61:6768-76
14 SehiffenbauerYS, MeirG, MaozM, et al.Gonadotropin stimulation of MLS human epithelial ovarian carcinoma cells augments cell adhesion mediated by CD44 and by alpha(v)-integrin.Gynecol Oncol, 2002, 84:296-302
15 Choi JH, Choi KC, AuerspergN, et al.Gonadotropins activate proteolysis and increase invasion through protein kinase A and phosphatidylinositol 3-kinase pathways in human epithelial ovarian cancer cells.Cancer Res, 2006, 66:3912-20
16 EmonsG, OrtmannO, TeiehertHM, et al.Luteinizing hormone-releasing hormone agonist triptorelin in combination with eytotoxie chemotheray in patients with advanced ovarian careinoma.Caneer, 1996, 78:1452-60
17黄春芳,刘东远,徐卫华,等.促卵泡激素对顺铂诱导卵巢癌细胞株凋亡的保护作用.中国医学科学院学,2003,25:443-6
18黄春芳,刘东远,沈铿.促卵泡刺激素抑制顺铂诱导的卵巢癌细胞凋亡.中国医学科学院学报,2003,25:447-50
19 Choi JH, Choi KC, Auersperg N, et al.E differential regulation of two forms of gonadotropin-releasing hormone messenger ribonueleie acid by gonadotropins in human immortalized ovarian surface epithelium and ovarian cancer cells.Endoer Relat Caneer, 2006, 13:641-51
20 Wang J, LuoF, LuJJ, et al.VEGF expression and enhanced production by gonadotropins in ovarian epithelial tumors.IntJ cancer, 2002, 97:163-7
21 Parrott JA, DoraiswamyV, KimG, et al.Expression and actions of both the folliele stimulating hormone receptor and the luteinizing hormone receptor in normal ovarian surface epithelium and ovarian cancer.Mol Cell Endoerinol, 2001, 172:213-22
22 Kraemer S, JaegerWH, LangN. Growth regulation effcets of gonadotropin induced steroidogenic response in human ovarian cancer.Anticancer Res, 2001, 21:2005-10
23 Weiss G, Skurnick JH, Goldsmith LT,et al Menopause and hypothalamic pituitary sensitivity to estrogen. JAMA 2004 292:2991-2996
24 Lu JJ, ZhengY, YuanJM, et al.Decreased luteinizing hormone receptor mRNA expression in human ovarian epithelial cancer. GynecolOncol, 2000,79:158-168
25 Wang J, Lin L, Parkash V, Schwartz PE.Quantitative analysis of follicle-stimulating hormone receptor in ovarian epithelial tumors: a novel approach to explain the field effect of ovarian cancer development in secondary Mullerian systems. Int J Cancer 2003 103:328-334
26 Choi KC, Kang SK, Tai CJ,et al. Follicle-stimulating hormone activates mitogen-activated protein kinase in preneoplastic and neoplastic ovarian surface epithelial cells. J Clin Endocrinol Metab 2002 87:2245-2253
27 Choi JH, Choi KC, Auersperg N, et al.Overexpression of follicle stimulating hormone receptor activates oncogenic pathways in preneoplastic ovarian surface epithelial cells. J Clin Endocrinol Metab2004 89:5508-5516
28 Zheng W, Lu J, Luo F,et al.Ovarian epithelial tumor growth Promotion by follicle-stumilating hormone and inhibition of theeffectby luteinzing hormone. GynecolOncol,2000,76:80-88
29 Viqar S, Gregory U,Samuel C, et al.Reproductive hormone induced, STAT3-mediated interleukin-6 action in normal and malignant human ovarian surface epithelial cells. Nat Cancer Inst,2002,94(8):617-629
30丰有吉,张惜阴,葛柏青,等.促性腺激素对人卵巢上皮性癌细胞的增殖作用.中华妇产科杂志,1996,31:166-8
31周抗美,朱铭伟,丰有吉.促卵泡激素结合片段对卵巢癌细胞株的作用.中华妇产科杂志,1999,34:726-5
32朱铭伟,吴乾渝,葛柏青,等.卵泡刺激素对卵巢癌细胞的增殖及CAI25Ⅱ抗原分泌的作用.中华妇产科杂志,1998,33:225-6
33 SyedV, UlinskiG, MokSC, et al.Expression of gonadotropin receptor and growth responses to key reproduetive hormones in normal and malignant human ovarian surface epithelial cells.Cancer Res, 2001, 61:6768-76
34 JIQ, LiuPl, ChenPK, et al. Follicle stimulating hormone induced growth promotion and gene expression profiles on ovarian surface epithelial cells.Int J Cancer, 2004, 112:803-14
35 Alam H, Maizels ET, Park Y, et al. Follicle-stimulating hormone activation of hypoxia-inducible factor-1 by the phosphatidylinositol 3-kinase/AKT/Ras homolog enriched in brain (Rheb)/mammalian target of rapamycin (mTOR) pathway is necessary for induction of select protein markers of follicular differentiation. J Biol Chem 2004; 279:19431-19440
36 Carvalho CR, Carvalheira JB, Lima MH, et al.Novel signal transduction pathway for luteinizing hormone and its interaction with insulin: activation of Janus kinase/signal transducer and activator of transcription and phosphoinositol 3-kinase/Akt pathways. Endocrinology 2003; 144:638-647
37 Meroni SB, Riera MF, Pellizzari EH, et al. FSH activates phosphati- dylinositol 3-kinase/protein kinase B signaling pathway in 20-day-oldSertoli cells independently of IGF-I. J Endocrinol 2004; 180:257-265
38 Bader AG, Kang S, Zhao L, et al.Oncogenic PI3K deregulates transcri- ption and translation. Nat Rev Cancer 2005 5: 921–929
39 Engelman JA, Luo J, Cantley LC.The evolution of phosphatidylinositol 3-kinases as regulators of growth and metabolism. Nat Rev Genet 2006 7: 606–619
40 Katso R, Okkenhaug K, Ahmadi K, et al. Cellular function of phosphoino- sitide 3-kinases: implications for development, homeostasis, and cancer. Annu Rev Cell Dev Biol 2001 17: 615-675
41 Parsons R. Phosphatidylinositol 3-kinase inhibitors are a triple threat to ovarian cancer. Clin Cancer Res 2005 11: 7965-7966
42 Vivanco I, Sawyers CL. The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat Rev Cancer 2002 2: 489-501
43 FresnoVaraJA, CasadoE, deCastroJ,et al.PI3K/Akt signalling Pathway and caneer. Cancer Treat Rev, 2004, 30(2):193-204
44 Testa JR, BellaeosaA.AKT plays a central role in tumorigenesis. Proc Nat Acad Sci USA.2001;98:10983-10985
45 HanadaM,FengJ,HenuningsBA.Structure, regulation and function of PKB/AKT-a major therapeutic target. Biochim Biophys Acta. 2004; 1697: 3216
46 Hill MM, Henunings BA. lnhibition of protein kinase B/Akt Implications for cancer therapy. Pharnlacol Ther.2002;93:243-251
47 Kermedy SG, Kandel ES, Cross TK, et al.Akt/Protein kinase B inhibits cell death by preventing the release of cytoehrome from mitochondria.Mol Cell Biol, 1999;19:5800-5810
48 GagnonV, St Germain ME, Parent S, et al.Akt activity in endometrial cancer cells:regulation of cell survival through cIAP-1.Int J Oncol. 2003; 23:803-810
49 Knuefermann C, Lu Y, Liu B, et al. HER2/PI3K/Akt activation leads to a multidrug resistance in hurnan breast adenocareinoma cells. oncogene. 2003;22:3205-3212
50 Brognard J, Clark AS, Ni Y, et al.Akt/protein kinase B is constitutively active in non-small cell lung cancer cells and promotes cellular survival and resistance to chemotherapy and radiation. Cancer Res. 2001; 61: 3986-3997
51 LinX, BohleAS, DohrmannP, et al.Over expression of phosphatidylin- ositol-3-kinase in human lung cancer. Langenbecks Aieh Surg, 2001, 386: 293-301
52 Sun M, Paeiga JE, Feldman RI, et al.Phosphatidylinositol-3-Ohkinase (PI3K)/AKT2, activated in breast cancer regulates and is induced by estrogen receptor (ER) via inieraetion between ER and PI3K.Can Res, 2001, 61:5985-5991
53 Gershtein ES, Scherbakov AM, Shatskaya VA, et al.Phosphatidylinositol3 -kinase/AKT signalling pathway components in human breast cancer: clinicopathological correlations. Anticancer Res, 2007, 27(4A):1777-82
54 Shukla S, Maclennan GT, Hartman DJ, et al.Activation of PI3K-Akt tignaling pathway promotes prostate cancer cell invasion. Int J Cancer, 2007,121(7):1424-32
55 Murakami D, Tsujitani S, Osaki T, et al.Expression of phosphorylated Akt (pAkt) in gastric carcinoma predicts prognosis and effieacy of chemo- therapy.GastrieCancer, 2007, 10(1):45-51
56 QianY, CoruxnL, MengQ, et al. PI3K induced actin filament remodeling through Akt and P7OS6KI:implication of essential role in cell migration. Am J Physiol Cell Physiol, 2004, 286(9):C153-C163
57 Park CM, Park MJ, Kwak HJ, et al. Ionizing radiation enhances matrix metallop roteinase-2 Secretion and invasion of Glioma cells through cre/epidermal growth factor receptor mediated P38/Akt and phosphatidy- linositol 3-kinase/Akt signaling pathways.Cancer Res, 2006, 66 (17): 8511 -8519
58 Kim CS, Vasko VV, Kato Y, et al.AKT activation promotes metastasis in amouse model offollieular thyroid careinoma[J].Endocrinology, 2005, 146 (6) 4456-4463
59 Romashkova, J.A. and Makarov, S.S. NF-kappaB is a target of AKT in antiapoptotic PDGF signalling. Nature, 1999 401,86-90
60 Chandrasekar, B., Bysani, S. and Mummidi,S. CXCL16 signals via Gi, phosphatidylinositol 3-kinase, Akt, I kappa B kinase and nuclear factor kappa B and induces cell-cell adhesion and aortic smooth muscle cell proliferation. J. Biol. Chem. 2004 279, 3188--3196
61 AldwinASJr.The NF-κB and lκB proteins:new diseoveries and insights. Armu Rev Immul, 1996, 14(2):649-683
62 Gustin JA, Ozes ON, Akca H,et al.Cell type-specific expression of the IkappaB kinases determines the significance of phosphatidylinositol 3 -kinase/A lot signaling to NF-kappaB activation. J Biol Chem, 2004, 279: 1615-1620
63 Joyee D, Albanese C, Steer J, et al.NF-kappaB and cell cycle regulation: the cyclin connection. Cytokine Growth Fator Rev, 2001, 12(l):73-90
64 Singh H, Sen R, Baltimore D, et al.A unclear factor that binds to a conserved sequence motif in transcriptional control elements of immuno- globulin genes.Nature, 1986, 319(6049):154-158
65 Escárcega RO, Fuentes-Alexandro S, García-Carrasco M et al.The transcription factor nuclear factor-kappa B and cancer. Clin Oncol (R Coll Radiol) 2007; 19: 154–61
66 Basseres DS, Baldwin AS.Nuclear factor- kappaB and inhibitor of kappaB kinase pathways in oncogenic initiation and progression. Oncogene 2006; 25: 6817-30
67 Levidou G, Korkolopoulou P, Nikiteas N et al. Expression of nuclear factor kappaB in human gastric carcinoma: Relationship with IkappaBa and prognostic significance. Virchows Arch 2007;450: 519–27
68 TangX, LiuD, ShishodiaS, et al.Nuclear factor-kappaB(NF-kappaB) is frequently expressed in lung cancer and preneoplastic lesions. Cancer 2006; 107: 2637–46
69 Yu LL, Yu HG, Yu JP et al. Nuclear factor-kappaB p65(RelA) transcription factor is constitutively activated in human colorectal carcinoma tissue.World J Gastroenterol 2004; 10:3255–60
70 Haffner MC, Berlato C, Doppler W.Exploiting our knowledge of NF-kappaB signaling for the treatment of mammary cancer. J Mammary Gland Biol Neoplasia 2006; 11: 63-73
71 Wu JT, Kral JG. The NF-kappaB/IkappaB signaling system: A molecular target in breast cancer therapy. J Surg Res 2005;123: 158-69
72 Cao Y, Karin M. NF-kappa B in mammary gland development and bast cancer. J Mammary Gland Biol Neoplasia 2003; 8: 215-23
73 Pikarsky E, Porat RM, Stein I, et al. NF-kappaB functions as a tumour promoter in inflammation-associated cancer. Nature 2004 431:461-466
74 Yoon50, Shin.5, Lee HJ, et al. Isoginkgetin inhibits tumor cell invasion by regulating phosphatidylinositol 3-kinase/Akt dependent matrix metallopr- oteinase-9 expression. Mol Caner Ther, 2006, 5(11):2666-2675
75 ChenY, LIR, WangR, et al. The signifieance of nuclear factor kappaB p65(NfkappaB p65)expression on the vaseular endothelial cells of rectum adnoeareinoma of human.Hua Xi Yi Ke Da Xue Xue Bao, 2001, 32(2):196-199
1 Astanehe A, Arenillas D, Wasserman WW, et al. Mechanisms un-derlyingp53 regulation of PIK3CA transcription in ovarian surface epithelium and in ovarian cancer.J Cell Sci,2008,121(pt 5):664-674
2 Woenckhaus J,Steger K,Sturm K,et al.Prognostic value of PIK3CA and phosphorylated AKT expression in ovarian cancer.Virchows Arch, 2007, 450(4):387-395
3 Graeff P, Crijns AP, Ten Hoor KA, et al. The ErbB signalling path-way:protein expression and prognostic value in epithelial ovarian cancer. Br J Cancer,2008,99(2):341-349
4 Isonishi S, Saitou M, Saitou M, et al. Differential regulation of thecytotoxicity activity of paclitaxel by orobol and platelet derived growth factor in human ovarian carcinoma cells.Oncol Rep,2007,18(1):195-201
5 Kim SH, Juhnn YS, Song YS.Akt involvement in paclitaxel chemoresi- stance of human ovarian cancer cells.Ann N Y Acad Sci,2007,1095:82-89
6 Yang X,Fraser M,Abedini MR,et al.Regulation of apoptosis-inducing factor-mediated,cisplatin-induced apoptosis by Akt.Br J Cancer, 2008, 98(4) :803-808
7 Patel BB, He YA, Li XM, et al.Molecular mechanisms of action ofimatinib mesylate in human ovarian cancer: a proteomic analysis. Cancer Genomics Proteomics,2008,5(3/4):137-149
8 Raynaud FI, Eccles S, Clarke PA, et al. Pharmacologic characterization of a potent inhibitor of class I phosphatidylinositide 3-kinases. Cancer Res, 2007, 67(12):5840-5850
9 Kawaguchi W, Itamochi H, Kigawa J, et al.Simultaneous inhibition of the mitogen-activated protein kinase and phosphatidyli-nositol 3′-kinase pathways enhances sensitivity to paclitaxel inovarian carcinoma.Cancer Sci,2007,98(12):2002-2008
10 Azzi A,Boscoboinik, D,HenseyC. The protein kinase C family.Eur J Biochem, 1992,208(3):547-557
11 Perletti GP, Majrras E, Coneari P, et al. PKC delta acts as a growth and tumor suppressor in rat clonic epithelial cellsyne GC, Marray AW.Evidence that protein kinase Cepsilon mediates phorbolester inhibition of calphostin C and tumor necrosis factor-alpha induced apoptosis in U937 histiocytic lymphoma cell. J Biol Chem, 1998; 273(37):24115-24121
12 Chamber TC, McAvoy EM, Jacobs JM, et al. Protein kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cell. Jbiol Chem, 1990,265(13):7679-7686.
13 Yu G, Ahmad S, Aquino A, et al. Transfection with protein kilnase C alpha confers increased multidrug resistance to MCF-7 cells expressing P-glycoprotein. Cancer commum, 1991,3(6):181-189
14 Choi KC, AuersPergN, Leung P. Mitogen-activated protein kinases in normal and (pre)neolastic ovarian surface epithelium. Reprod Biol Endocrinol.2003;7(1) :71-78
15韦淑琴,隋丽华,戚基萍等.卵巢上皮性肿瘤组织中ERK的表达与活化.哈尔滨医科大学学报.2004;38(3):233-236
16 HsuCY, BristowR, Cha MS, et al. Characterization of active mitogen-activated protein kinase in ovarian serous carcinomas. Clin Cancer Res. 2004; 10(19): 6432-6436
17 PohlG, HoCL, KurmanR, et al. Inaetivation of the mitogen-aetivated Protein kinase Pathway as a Potential target-based therapy in ovarian serous tumors with KRAS or BRAF mutations.Cancer Res. 2005; 65(5): 1994-2000
18 Choi KC, Auersperg N, Leung P. Mitogen-activated protein kinases in normal and (pre)neoplastic ovarian surface epithelium. Reprod Biol Endocrinol. 2003; 7(1): 71-78
19 Choi JH, Choi KC,Auersperg N. et al.Gonadotropins upregulate the epidermal growth factor receptor through activation of mitogen activated protein kinases and phosphatidylinositol 3-kinaes in human ovarian surface epithelial cells. Endocrine-arelated Cancer. 2005; 12:407-421
20 FrasehSC, NickJA, Fadok VA, et al. p38 mitogen activated protein kinase dependent and independent intracellular signal transduction pathways leading to a poptosis in human neutrophils.Biol Chem. 1998; 273 (14):8389-8397
21 Li G,Xiang Y,Sabapathy K, et al. Anapoptotic signaling pathway in the interferon antiviral response mediated by Rnase L and c-Jun NH2-terminal kinase. Biol Chem.2004;279(2):1123-1132
22 Kim KY,Choi KC,Park SH, et al. Extracellular signal-regulated protein kinase,but not c-Jun N-terminal kinase,is activated by type II gonadotropin-releasing hormone involved in the inhibition of ovarian cancer cell proliferation. Clin Endocrinol Metab. 2005;90(3):1670-1677
23 Kim KY,Choi KC,Park SH, et al. Type II gonadotropin-releasing hormone stimulates p38 mitogen-activated protein kinase and apoptosis in ovarian cancer cells. Clin Endocrinol Metab.2004;89(6):3020-3026
24 Klampfer L. Signal transducers and activators of transcription(STATs): Novel targets of chemopreventive and chemotherarpeutic drugs . Curr Cancer Drug Targets,2006,6(2):107-121
25 Rosen DG, Mercado-Uribe I, Yang G, et al. The role of constitutively active signal transducer and activator of transcription in ovarian tumorigenesis and prognosis. Cancer, 2006,107(11):2730-2740
26 Xie TX,Wei D,Liu M,et al.Stat3 activation regulates the expression of matrixmetallop roteinase-2 and tumor invasion and metastasis. Oncogene, 2004, 23(20):3550-3560
27 Rivat C, De Wever O, Bruyneel E, et al.Disruption of STAT3 signaling leeds to tumor cell invasion through alteration of homotypic cell-cell adhersion complexes.Oncogene,2004,23(19):3317-3327
28 Vanasse GJ,Winn RK,Rodov S,et al.Bcl-2 overexpression leads to increases in suppressor of cytokine signaling23 expression in B cells and de novo follicular lymphoma.Mol Cancer Res,2004,2(11):620-631
29 Silver DL,Naora H,Liu J,et al.Activated signal transducer and activator of transcription(STAT)3:localization in focal adhesions and function in ovarian cancer cell motility. Cancer Res,2004,64(10);3550-3558
30 Yahata Y,Shirakata Y,Tokumaru S,et al.Nuclear translocation of phosph- orylated STAT3 is essential for vascular endothelial growth factorinduced human dermal microvascular endothelial cell migration and tube formation.J Biol Chem,2003,278(41):40026-40031
31 Rosen DG, Mercado-Uribe I, Yang G, et al.The role of constitutively active signal transducer and activator of transcription 3 in ovarian tumorigenesis and prognosis.Cancer, 2006,107(11):2730-2740
32 Collins FS, Green ED, Guttmacher AE, et al. A vision for the future of genomics research. Nature. 2003 Apr 24;422(6934):835-47
2 Keith D, Edmonds, Dewhurst Text book of Obstetrics & Gynaecology. UK:Black well Publishing, seventh edition.2007.625
3 Christine H, Holschneicerm MD, Jonathan S, et al.Advances in the understanding and treatment of ovarian cancer. Brmeno-pause Soc, 2005, 11(2):66-71
4 Boso CK. Role of nerve growth factor and FSH receptor in epithelial ovarian cancer. Reprod Biomed Online,2005 11(2):194-197
5 Choi JH, Wong AS, Endocr Rev.Gonadotropins and Ovarian Cancer 2007 Jun; 28(4):440-61
6 Huang Y, Hua K.Activation of the PI3K/AKT pathway mediates FSH stimulated VEGF expression in ovarian serous cystadenocarcinoma. CellRes. 2008 Jul;18(7):780-91
7 Min C, Eddy SF. NF-kappaB and epithelial to mesenchymal transition of cancer. J Cell Biochem. 2008 Jun 1;104(3):733-44
8 Wang Y, Chan S, Tsang BK.Involvement of inhibitory nuclear factor kappaB activation in the gonadotropic regulation of X-linked inhibitor of apoptosis expression during ovarian follicular development in vitro. Endocrinology. 2002 Jul;143(7):2732-40
9 Bani MR, Nieoletti Ml, Aikarouf NW, et al.Gene expression correlating with response to paclitaxel in ovarian carcinoma xenografts. Mol Cancer Ther, 2004,3(2):111-121
10丰有吉,张惜阴,葛柏青,等.促性腺激素对人卵巢上皮性癌细胞的增殖作用.中华妇产科杂志,1996,31:166-8
11 Maruuehi T, Sugiyama T,Kataoka A,et al.Effects of a gonadotropin releasing hormone agonist on rat ovarian adenocarcinoma cell lines in vitro and in vivo. JPnJ Cancer Res,1998,89:977-83
12朱铭伟,吴乾渝,葛柏青等卵泡刺激素对卵巢癌细胞的增殖及CA125Ⅱ抗原分泌的作用.中华妇产科杂志,,1998,33:225-6
13 Syedy, UlinskiG, MokSC, et al.Expression of gonadotropin receptor and growth responses to key reproduetive hormones in normal and malignant human ovarian surface epithelial cells.Cancer Res, 2001, 61:6768-76
14 SehiffenbauerYS, MeirG, MaozM, et al.Gonadotropin stimulation of MLS human epithelial ovarian carcinoma cells augments cell adhesion mediated by CD44 and by alpha(v)-integrin.Gynecol Oncol, 2002, 84:296-302
15 Choi JH, Choi KC, AuerspergN, et al.Gonadotropins activate proteolysis and increase invasion through protein kinase A and phosphatidylinositol 3-kinase pathways in human epithelial ovarian cancer cells.Cancer Res, 2006, 66:3912-20
16 EmonsG, OrtmannO, TeiehertHM, et al.Luteinizing hormone-releasing hormone agonist triptorelin in combination with eytotoxie chemotheray in patients with advanced ovarian careinoma.Caneer, 1996, 78:1452-60
17黄春芳,刘东远,徐卫华,等.促卵泡激素对顺铂诱导卵巢癌细胞株凋亡的保护作用.中国医学科学院学,2003,25:443-6
18黄春芳,刘东远,沈铿.促卵泡刺激素抑制顺铂诱导的卵巢癌细胞凋亡.中国医学科学院学报,2003,25:447-50
19 Choi JH, Choi KC, Auersperg N, et al.E differential regulation of two forms of gonadotropin-releasing hormone messenger ribonueleie acid by gonadotropins in human immortalized ovarian surface epithelium and ovarian cancer cells.Endoer Relat Caneer, 2006, 13:641-51
20 Wang J, LuoF, LuJJ, et al.VEGF expression and enhanced production by gonadotropins in ovarian epithelial tumors.IntJ cancer, 2002, 97:163-7
21 Parrott JA, DoraiswamyV, KimG, et al.Expression and actions of both the folliele stimulating hormone receptor and the luteinizing hormone receptor in normal ovarian surface epithelium and ovarian cancer.Mol Cell Endoerinol, 2001, 172:213-22
22 Kraemer S, JaegerWH, LangN. Growth regulation effcets of gonadotropin induced steroidogenic response in human ovarian cancer.Anticancer Res, 2001, 21:2005-10
23 Weiss G, Skurnick JH, Goldsmith LT,et al Menopause and hypothalamic pituitary sensitivity to estrogen. JAMA 2004 292:2991-2996
24 Lu JJ, ZhengY, YuanJM, et al.Decreased luteinizing hormone receptor mRNA expression in human ovarian epithelial cancer. GynecolOncol, 2000,79:158-168
25 Wang J, Lin L, Parkash V, Schwartz PE.Quantitative analysis of follicle-stimulating hormone receptor in ovarian epithelial tumors: a novel approach to explain the field effect of ovarian cancer development in secondary Mullerian systems. Int J Cancer 2003 103:328-334
26 Choi KC, Kang SK, Tai CJ,et al. Follicle-stimulating hormone activates mitogen-activated protein kinase in preneoplastic and neoplastic ovarian surface epithelial cells. J Clin Endocrinol Metab 2002 87:2245-2253
27 Choi JH, Choi KC, Auersperg N, et al.Overexpression of follicle stimulating hormone receptor activates oncogenic pathways in preneoplastic ovarian surface epithelial cells. J Clin Endocrinol Metab2004 89:5508-5516
28 Zheng W, Lu J, Luo F,et al.Ovarian epithelial tumor growth Promotion by follicle-stumilating hormone and inhibition of theeffectby luteinzing hormone. GynecolOncol,2000,76:80-88
29 Viqar S, Gregory U,Samuel C, et al.Reproductive hormone induced, STAT3-mediated interleukin-6 action in normal and malignant human ovarian surface epithelial cells. Nat Cancer Inst,2002,94(8):617-629
30丰有吉,张惜阴,葛柏青,等.促性腺激素对人卵巢上皮性癌细胞的增殖作用.中华妇产科杂志,1996,31:166-8
31周抗美,朱铭伟,丰有吉.促卵泡激素结合片段对卵巢癌细胞株的作用.中华妇产科杂志,1999,34:726-5
32朱铭伟,吴乾渝,葛柏青,等.卵泡刺激素对卵巢癌细胞的增殖及CAI25Ⅱ抗原分泌的作用.中华妇产科杂志,1998,33:225-6
33 SyedV, UlinskiG, MokSC, et al.Expression of gonadotropin receptor and growth responses to key reproduetive hormones in normal and malignant human ovarian surface epithelial cells.Cancer Res, 2001, 61:6768-76
34 JIQ, LiuPl, ChenPK, et al. Follicle stimulating hormone induced growth promotion and gene expression profiles on ovarian surface epithelial cells.Int J Cancer, 2004, 112:803-14
35 Alam H, Maizels ET, Park Y, et al. Follicle-stimulating hormone activation of hypoxia-inducible factor-1 by the phosphatidylinositol 3-kinase/AKT/Ras homolog enriched in brain (Rheb)/mammalian target of rapamycin (mTOR) pathway is necessary for induction of select protein markers of follicular differentiation. J Biol Chem 2004; 279:19431-19440
36 Carvalho CR, Carvalheira JB, Lima MH, et al.Novel signal transduction pathway for luteinizing hormone and its interaction with insulin: activation of Janus kinase/signal transducer and activator of transcription and phosphoinositol 3-kinase/Akt pathways. Endocrinology 2003; 144:638-647
37 Meroni SB, Riera MF, Pellizzari EH, et al. FSH activates phosphati- dylinositol 3-kinase/protein kinase B signaling pathway in 20-day-oldSertoli cells independently of IGF-I. J Endocrinol 2004; 180:257-265
38 Bader AG, Kang S, Zhao L, et al.Oncogenic PI3K deregulates transcri- ption and translation. Nat Rev Cancer 2005 5: 921–929
39 Engelman JA, Luo J, Cantley LC.The evolution of phosphatidylinositol 3-kinases as regulators of growth and metabolism. Nat Rev Genet 2006 7: 606–619
40 Katso R, Okkenhaug K, Ahmadi K, et al. Cellular function of phosphoino- sitide 3-kinases: implications for development, homeostasis, and cancer. Annu Rev Cell Dev Biol 2001 17: 615-675
41 Parsons R. Phosphatidylinositol 3-kinase inhibitors are a triple threat to ovarian cancer. Clin Cancer Res 2005 11: 7965-7966
42 Vivanco I, Sawyers CL. The phosphatidylinositol 3-Kinase AKT pathway in human cancer. Nat Rev Cancer 2002 2: 489-501
43 FresnoVaraJA, CasadoE, deCastroJ,et al.PI3K/Akt signalling Pathway and caneer. Cancer Treat Rev, 2004, 30(2):193-204
44 Testa JR, BellaeosaA.AKT plays a central role in tumorigenesis. Proc Nat Acad Sci USA.2001;98:10983-10985
45 HanadaM,FengJ,HenuningsBA.Structure, regulation and function of PKB/AKT-a major therapeutic target. Biochim Biophys Acta. 2004; 1697: 3216
46 Hill MM, Henunings BA. lnhibition of protein kinase B/Akt Implications for cancer therapy. Pharnlacol Ther.2002;93:243-251
47 Kermedy SG, Kandel ES, Cross TK, et al.Akt/Protein kinase B inhibits cell death by preventing the release of cytoehrome from mitochondria.Mol Cell Biol, 1999;19:5800-5810
48 GagnonV, St Germain ME, Parent S, et al.Akt activity in endometrial cancer cells:regulation of cell survival through cIAP-1.Int J Oncol. 2003; 23:803-810
49 Knuefermann C, Lu Y, Liu B, et al. HER2/PI3K/Akt activation leads to a multidrug resistance in hurnan breast adenocareinoma cells. oncogene. 2003;22:3205-3212
50 Brognard J, Clark AS, Ni Y, et al.Akt/protein kinase B is constitutively active in non-small cell lung cancer cells and promotes cellular survival and resistance to chemotherapy and radiation. Cancer Res. 2001; 61: 3986-3997
51 LinX, BohleAS, DohrmannP, et al.Over expression of phosphatidylin- ositol-3-kinase in human lung cancer. Langenbecks Aieh Surg, 2001, 386: 293-301
52 Sun M, Paeiga JE, Feldman RI, et al.Phosphatidylinositol-3-Ohkinase (PI3K)/AKT2, activated in breast cancer regulates and is induced by estrogen receptor (ER) via inieraetion between ER and PI3K.Can Res, 2001, 61:5985-5991
53 Gershtein ES, Scherbakov AM, Shatskaya VA, et al.Phosphatidylinositol3 -kinase/AKT signalling pathway components in human breast cancer: clinicopathological correlations. Anticancer Res, 2007, 27(4A):1777-82
54 Shukla S, Maclennan GT, Hartman DJ, et al.Activation of PI3K-Akt tignaling pathway promotes prostate cancer cell invasion. Int J Cancer, 2007,121(7):1424-32
55 Murakami D, Tsujitani S, Osaki T, et al.Expression of phosphorylated Akt (pAkt) in gastric carcinoma predicts prognosis and effieacy of chemo- therapy.GastrieCancer, 2007, 10(1):45-51
56 QianY, CoruxnL, MengQ, et al. PI3K induced actin filament remodeling through Akt and P7OS6KI:implication of essential role in cell migration. Am J Physiol Cell Physiol, 2004, 286(9):C153-C163
57 Park CM, Park MJ, Kwak HJ, et al. Ionizing radiation enhances matrix metallop roteinase-2 Secretion and invasion of Glioma cells through cre/epidermal growth factor receptor mediated P38/Akt and phosphatidy- linositol 3-kinase/Akt signaling pathways.Cancer Res, 2006, 66 (17): 8511 -8519
58 Kim CS, Vasko VV, Kato Y, et al.AKT activation promotes metastasis in amouse model offollieular thyroid careinoma[J].Endocrinology, 2005, 146 (6) 4456-4463
59 Romashkova, J.A. and Makarov, S.S. NF-kappaB is a target of AKT in antiapoptotic PDGF signalling. Nature, 1999 401,86-90
60 Chandrasekar, B., Bysani, S. and Mummidi,S. CXCL16 signals via Gi, phosphatidylinositol 3-kinase, Akt, I kappa B kinase and nuclear factor kappa B and induces cell-cell adhesion and aortic smooth muscle cell proliferation. J. Biol. Chem. 2004 279, 3188--3196
61 AldwinASJr.The NF-κB and lκB proteins:new diseoveries and insights. Armu Rev Immul, 1996, 14(2):649-683
62 Gustin JA, Ozes ON, Akca H,et al.Cell type-specific expression of the IkappaB kinases determines the significance of phosphatidylinositol 3 -kinase/A lot signaling to NF-kappaB activation. J Biol Chem, 2004, 279: 1615-1620
63 Joyee D, Albanese C, Steer J, et al.NF-kappaB and cell cycle regulation: the cyclin connection. Cytokine Growth Fator Rev, 2001, 12(l):73-90
64 Singh H, Sen R, Baltimore D, et al.A unclear factor that binds to a conserved sequence motif in transcriptional control elements of immuno- globulin genes.Nature, 1986, 319(6049):154-158
65 Escárcega RO, Fuentes-Alexandro S, García-Carrasco M et al.The transcription factor nuclear factor-kappa B and cancer. Clin Oncol (R Coll Radiol) 2007; 19: 154–61
66 Basseres DS, Baldwin AS.Nuclear factor- kappaB and inhibitor of kappaB kinase pathways in oncogenic initiation and progression. Oncogene 2006; 25: 6817-30
67 Levidou G, Korkolopoulou P, Nikiteas N et al. Expression of nuclear factor kappaB in human gastric carcinoma: Relationship with IkappaBa and prognostic significance. Virchows Arch 2007;450: 519–27
68 TangX, LiuD, ShishodiaS, et al.Nuclear factor-kappaB(NF-kappaB) is frequently expressed in lung cancer and preneoplastic lesions. Cancer 2006; 107: 2637–46
69 Yu LL, Yu HG, Yu JP et al. Nuclear factor-kappaB p65(RelA) transcription factor is constitutively activated in human colorectal carcinoma tissue.World J Gastroenterol 2004; 10:3255–60
70 Haffner MC, Berlato C, Doppler W.Exploiting our knowledge of NF-kappaB signaling for the treatment of mammary cancer. J Mammary Gland Biol Neoplasia 2006; 11: 63-73
71 Wu JT, Kral JG. The NF-kappaB/IkappaB signaling system: A molecular target in breast cancer therapy. J Surg Res 2005;123: 158-69
72 Cao Y, Karin M. NF-kappa B in mammary gland development and bast cancer. J Mammary Gland Biol Neoplasia 2003; 8: 215-23
73 Pikarsky E, Porat RM, Stein I, et al. NF-kappaB functions as a tumour promoter in inflammation-associated cancer. Nature 2004 431:461-466
74 Yoon50, Shin.5, Lee HJ, et al. Isoginkgetin inhibits tumor cell invasion by regulating phosphatidylinositol 3-kinase/Akt dependent matrix metallopr- oteinase-9 expression. Mol Caner Ther, 2006, 5(11):2666-2675
75 ChenY, LIR, WangR, et al. The signifieance of nuclear factor kappaB p65(NfkappaB p65)expression on the vaseular endothelial cells of rectum adnoeareinoma of human.Hua Xi Yi Ke Da Xue Xue Bao, 2001, 32(2):196-199
1 Astanehe A, Arenillas D, Wasserman WW, et al. Mechanisms un-derlyingp53 regulation of PIK3CA transcription in ovarian surface epithelium and in ovarian cancer.J Cell Sci,2008,121(pt 5):664-674
2 Woenckhaus J,Steger K,Sturm K,et al.Prognostic value of PIK3CA and phosphorylated AKT expression in ovarian cancer.Virchows Arch, 2007, 450(4):387-395
3 Graeff P, Crijns AP, Ten Hoor KA, et al. The ErbB signalling path-way:protein expression and prognostic value in epithelial ovarian cancer. Br J Cancer,2008,99(2):341-349
4 Isonishi S, Saitou M, Saitou M, et al. Differential regulation of thecytotoxicity activity of paclitaxel by orobol and platelet derived growth factor in human ovarian carcinoma cells.Oncol Rep,2007,18(1):195-201
5 Kim SH, Juhnn YS, Song YS.Akt involvement in paclitaxel chemoresi- stance of human ovarian cancer cells.Ann N Y Acad Sci,2007,1095:82-89
6 Yang X,Fraser M,Abedini MR,et al.Regulation of apoptosis-inducing factor-mediated,cisplatin-induced apoptosis by Akt.Br J Cancer, 2008, 98(4) :803-808
7 Patel BB, He YA, Li XM, et al.Molecular mechanisms of action ofimatinib mesylate in human ovarian cancer: a proteomic analysis. Cancer Genomics Proteomics,2008,5(3/4):137-149
8 Raynaud FI, Eccles S, Clarke PA, et al. Pharmacologic characterization of a potent inhibitor of class I phosphatidylinositide 3-kinases. Cancer Res, 2007, 67(12):5840-5850
9 Kawaguchi W, Itamochi H, Kigawa J, et al.Simultaneous inhibition of the mitogen-activated protein kinase and phosphatidyli-nositol 3′-kinase pathways enhances sensitivity to paclitaxel inovarian carcinoma.Cancer Sci,2007,98(12):2002-2008
10 Azzi A,Boscoboinik, D,HenseyC. The protein kinase C family.Eur J Biochem, 1992,208(3):547-557
11 Perletti GP, Majrras E, Coneari P, et al. PKC delta acts as a growth and tumor suppressor in rat clonic epithelial cellsyne GC, Marray AW.Evidence that protein kinase Cepsilon mediates phorbolester inhibition of calphostin C and tumor necrosis factor-alpha induced apoptosis in U937 histiocytic lymphoma cell. J Biol Chem, 1998; 273(37):24115-24121
12 Chamber TC, McAvoy EM, Jacobs JM, et al. Protein kinase C phosphorylates P-glycoprotein in multidrug resistant human KB carcinoma cell. Jbiol Chem, 1990,265(13):7679-7686.
13 Yu G, Ahmad S, Aquino A, et al. Transfection with protein kilnase C alpha confers increased multidrug resistance to MCF-7 cells expressing P-glycoprotein. Cancer commum, 1991,3(6):181-189
14 Choi KC, AuersPergN, Leung P. Mitogen-activated protein kinases in normal and (pre)neolastic ovarian surface epithelium. Reprod Biol Endocrinol.2003;7(1) :71-78
15韦淑琴,隋丽华,戚基萍等.卵巢上皮性肿瘤组织中ERK的表达与活化.哈尔滨医科大学学报.2004;38(3):233-236
16 HsuCY, BristowR, Cha MS, et al. Characterization of active mitogen-activated protein kinase in ovarian serous carcinomas. Clin Cancer Res. 2004; 10(19): 6432-6436
17 PohlG, HoCL, KurmanR, et al. Inaetivation of the mitogen-aetivated Protein kinase Pathway as a Potential target-based therapy in ovarian serous tumors with KRAS or BRAF mutations.Cancer Res. 2005; 65(5): 1994-2000
18 Choi KC, Auersperg N, Leung P. Mitogen-activated protein kinases in normal and (pre)neoplastic ovarian surface epithelium. Reprod Biol Endocrinol. 2003; 7(1): 71-78
19 Choi JH, Choi KC,Auersperg N. et al.Gonadotropins upregulate the epidermal growth factor receptor through activation of mitogen activated protein kinases and phosphatidylinositol 3-kinaes in human ovarian surface epithelial cells. Endocrine-arelated Cancer. 2005; 12:407-421
20 FrasehSC, NickJA, Fadok VA, et al. p38 mitogen activated protein kinase dependent and independent intracellular signal transduction pathways leading to a poptosis in human neutrophils.Biol Chem. 1998; 273 (14):8389-8397
21 Li G,Xiang Y,Sabapathy K, et al. Anapoptotic signaling pathway in the interferon antiviral response mediated by Rnase L and c-Jun NH2-terminal kinase. Biol Chem.2004;279(2):1123-1132
22 Kim KY,Choi KC,Park SH, et al. Extracellular signal-regulated protein kinase,but not c-Jun N-terminal kinase,is activated by type II gonadotropin-releasing hormone involved in the inhibition of ovarian cancer cell proliferation. Clin Endocrinol Metab. 2005;90(3):1670-1677
23 Kim KY,Choi KC,Park SH, et al. Type II gonadotropin-releasing hormone stimulates p38 mitogen-activated protein kinase and apoptosis in ovarian cancer cells. Clin Endocrinol Metab.2004;89(6):3020-3026
24 Klampfer L. Signal transducers and activators of transcription(STATs): Novel targets of chemopreventive and chemotherarpeutic drugs . Curr Cancer Drug Targets,2006,6(2):107-121
25 Rosen DG, Mercado-Uribe I, Yang G, et al. The role of constitutively active signal transducer and activator of transcription in ovarian tumorigenesis and prognosis. Cancer, 2006,107(11):2730-2740
26 Xie TX,Wei D,Liu M,et al.Stat3 activation regulates the expression of matrixmetallop roteinase-2 and tumor invasion and metastasis. Oncogene, 2004, 23(20):3550-3560
27 Rivat C, De Wever O, Bruyneel E, et al.Disruption of STAT3 signaling leeds to tumor cell invasion through alteration of homotypic cell-cell adhersion complexes.Oncogene,2004,23(19):3317-3327
28 Vanasse GJ,Winn RK,Rodov S,et al.Bcl-2 overexpression leads to increases in suppressor of cytokine signaling23 expression in B cells and de novo follicular lymphoma.Mol Cancer Res,2004,2(11):620-631
29 Silver DL,Naora H,Liu J,et al.Activated signal transducer and activator of transcription(STAT)3:localization in focal adhesions and function in ovarian cancer cell motility. Cancer Res,2004,64(10);3550-3558
30 Yahata Y,Shirakata Y,Tokumaru S,et al.Nuclear translocation of phosph- orylated STAT3 is essential for vascular endothelial growth factorinduced human dermal microvascular endothelial cell migration and tube formation.J Biol Chem,2003,278(41):40026-40031
31 Rosen DG, Mercado-Uribe I, Yang G, et al.The role of constitutively active signal transducer and activator of transcription 3 in ovarian tumorigenesis and prognosis.Cancer, 2006,107(11):2730-2740
32 Collins FS, Green ED, Guttmacher AE, et al. A vision for the future of genomics research. Nature. 2003 Apr 24;422(6934):835-47