FSH对卵巢癌细胞增殖、侵袭作用的影响及可能的机制
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摘要
目的:探讨卵泡刺激素(FSH)对卵巢癌细胞增殖、侵袭作用的影响及可能的机制。方法:(1)将购买自北京大学人民医院妇产科实验室的进入生长对数期的卵巢癌SKOV-3细胞与浓度为0、10、20、40、80 m IU·ml~(-1)的FSH液共培养,同时设立对照组(只加培养基,不加FSH),采用MMT法观察不同作用时间各组细胞的增殖情况;(2)将SKOV-3细胞分为空白组和FSH(40 m IU·ml~(-1))干预组共培养48 h,Wester-blotting法检测各组中FSH受体(FSHR)、磷酸化蛋白激酶B(p AKT)、E-cadherin蛋白的表达水平,采用Transwell小室测定细胞侵袭转移能力。结果:在共培养12、24、48、72 h时间点,FSH浓度10、20、40、80 m IU·ml~(-1)组的SKOV-3细胞增殖率显著高于0 m IU·ml~(-1)组(P<0.05);FSH浓度40 m IU·ml~(-1)组的SKOV-3细胞增殖率在共培养12、24、48、72 h时间点均显著高于FSH浓度10、20、80 m IU·ml~(-1)组(P<0.05),且在共培养48 h时达最大值;在共培养48 h后,FSH干预组的SKOV-3细胞上清液中p AKT蛋白水平显著高于空白组(P<0.05),E-cadherin蛋白水平显著低于空白组(P<0.05),两组间SKOV-3细胞上清液中FSHR蛋白表达水平差异无统计学意义(P>0.05);在共培养48 h后,FSH干预组的SKOV-3细胞穿过基质膜的平均细胞数显著高于空白组(P<0.05)。结论:FSH能促进卵巢癌细胞增殖,提高其侵袭能力,作用机制与降低E-cadherin蛋白水平、提高p AKT蛋白表达有关。
Objective: To investigate the mechanism of follicle stimulating hormone( FSH) in promoting the proliferation and invasion of ovarian cancer cells. Methods:( 1) SKOV-3 cells purchased from the Department of Obstetrics and Gynecology of Peking University People's Hospital Laboratory entered the logarithmic growth phase were divided into 0,10,20,40,80 m IU · ml~(-1) FSH coculture group. MMT method was used to observed the proliferation of cells in the groups at different action time;( 2) SKOV-3 cells were divided into control group and FSH( 40 m IU·ml~(-1)) intervention group,after cocultured 48 h,FSH receptors( FSHR),phosphorylated protein kinase B( p AKT),the expression level of E-cadherin protein were detected by Wester-blotting. Cell invasion and metastasis were determined by Transwell. Results: At the time of cocultured 12,24,48 and 72 h,the proliferation rate of SKOV-3 cells was significantly higher in FSH concentration 10,20,40 and 80 m IU·ml~(-1) four groups than in 0 m IU·ml~(-1) group( P < 0. 05). The cell proliferation rate of SKOV-3 cells in FSH 40 m IU·ml~(-1) group at cocultured 12,24,48,72 h time points were significantly higher than those of the FSH 10,20 and 80 m IU·ml~(-1) groups( P < 0. 05),and the maximum was reached at cocultured 48 h. After cocultured 48 h,the level of p AKT protein in the supernatant of SKOV-3 cells in FSH intervention group was significantly higher than that in the blank group( P < 0. 05); the level of E-cadherin protein was significantly lower than that of the blank group( P <0. 05); there was no significant difference in the expression of FSHR protein in the supernatant of SKOV-3 cells between the two groups( P > 0. 05). After cocultured 48 h,the average number of SKOV-3 cells passing through the stroma membrane in the FSH intervention group was significantly higher than that in the blank group( P <0. 05). Conclusion: FSH can promote the proliferation and invasion of ovarian cancer cells,the mechanism is reducing the level of E-cadherin protein and improving the expression of p AKT protein.
Objective: To investigate the mechanism of follicle stimulating hormone( FSH) in promoting the proliferation and invasion of ovarian cancer cells. Methods:( 1) SKOV-3 cells purchased from the Department of Obstetrics and Gynecology of Peking University People's Hospital Laboratory entered the logarithmic growth phase were divided into 0,10,20,40,80 m IU · ml~(-1) FSH coculture group. MMT method was used to observed the proliferation of cells in the groups at different action time;( 2) SKOV-3 cells were divided into control group and FSH( 40 m IU·ml~(-1)) intervention group,after cocultured 48 h,FSH receptors( FSHR),phosphorylated protein kinase B( p AKT),the expression level of E-cadherin protein were detected by Wester-blotting. Cell invasion and metastasis were determined by Transwell. Results: At the time of cocultured 12,24,48 and 72 h,the proliferation rate of SKOV-3 cells was significantly higher in FSH concentration 10,20,40 and 80 m IU·ml~(-1) four groups than in 0 m IU·ml~(-1) group( P < 0. 05). The cell proliferation rate of SKOV-3 cells in FSH 40 m IU·ml~(-1) group at cocultured 12,24,48,72 h time points were significantly higher than those of the FSH 10,20 and 80 m IU·ml~(-1) groups( P < 0. 05),and the maximum was reached at cocultured 48 h. After cocultured 48 h,the level of p AKT protein in the supernatant of SKOV-3 cells in FSH intervention group was significantly higher than that in the blank group( P < 0. 05); the level of E-cadherin protein was significantly lower than that of the blank group( P <0. 05); there was no significant difference in the expression of FSHR protein in the supernatant of SKOV-3 cells between the two groups( P > 0. 05). After cocultured 48 h,the average number of SKOV-3 cells passing through the stroma membrane in the FSH intervention group was significantly higher than that in the blank group( P <0. 05). Conclusion: FSH can promote the proliferation and invasion of ovarian cancer cells,the mechanism is reducing the level of E-cadherin protein and improving the expression of p AKT protein.
引文
[1]刘梦娜,谢静燕,赵树立.雌激素受体在卵巢癌研究中的进展[J].东南大学学报:医学版,2014,33(2):215-218.
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[9]赵一兵,戴志琴,沈杨.卵巢癌初治后阴道残端复发灶切除9例分析[J].现代医学,2015,43(10):1285-1286.
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[11]王昊,李佩玲.肿瘤干细胞标志物在卵巢癌中应用研究进展[J].中华实用诊断与治疗杂志,2015,29(9):840-841.
[12]张先华.血清HE4和CA125检测及ROMA模型在恶性卵巢癌诊断中的应用[J].实用癌症杂志,2015,31(1):17-19.
[13]张婧,金海艳,金丽仙.彩超诊断与血清CA125检测联用对卵巢癌的诊疗价值探究[J].中国妇幼保健,2015,30(32):5684-5686.
[14]赵鑫鑫,刘磊,张箴波.Nanog介导FSH诱导卵巢癌转移和侵袭的研究[J].现代妇产科进展,2015,24(5):321-324.
[15]徐春琳,路晓琳,闫晓楠.PI3K/Akt/NF-κB信号通路在FSH促进卵巢癌细胞增殖与侵袭中的作用[J].中华妇产科杂志,2012,47(2):134-138.
[2]田红,于鹏,吴小茗.卵巢癌的治疗药物研究进展[J].现代药物与临床,2015,36(1):103-107.
[3]WANG Z H,XU C J.Research progress of MicroRNA in early detection of ovarian cancer[J].中华医学杂志:英文版,2015,128(24):3363-3370.
[4]YANG L,HUANG S.Research progress in the treatment of ovarian cancer[J].China Medical Herald,2015,30(2):104-115.
[5]黄平,张立民,黄冬冬.缺氧诱导因子1α在促卵泡激素促进卵巢癌SKOV3细胞增殖与侵袭中的作用[J].河北医科大学学报,2016,37(4):450-453.
[6]刘晓梅,郑桂英.卵泡刺激素及其受体对卵巢癌细胞株的增殖作用[J].实用药物与临床,2015,18(4):398-402.
[7]BHARTIYA D,SINGH J.FSH-FSHR3-stem cells in ovary surface epithelium:basis for adult ovarian biology,failure,aging,and cancer[J].Reproduction,2015,149(1):35-48.
[8]LIU L,ZHANG J,FANG C,et al.OCT4 mediates FSH-induced epithelial-mesenchymal transition and invasion through the ERK1/2 signaling pathway in epithelial ovarian cancer[J].Biochem Bioph Res Co,2015,461(3):525-532.
[9]赵一兵,戴志琴,沈杨.卵巢癌初治后阴道残端复发灶切除9例分析[J].现代医学,2015,43(10):1285-1286.
[10]张苗,刘艳,张洁.新辅助化疗在治疗晚期卵巢癌中的应用进展[J].医学综述,2015,21(24):4483-4485.
[11]王昊,李佩玲.肿瘤干细胞标志物在卵巢癌中应用研究进展[J].中华实用诊断与治疗杂志,2015,29(9):840-841.
[12]张先华.血清HE4和CA125检测及ROMA模型在恶性卵巢癌诊断中的应用[J].实用癌症杂志,2015,31(1):17-19.
[13]张婧,金海艳,金丽仙.彩超诊断与血清CA125检测联用对卵巢癌的诊疗价值探究[J].中国妇幼保健,2015,30(32):5684-5686.
[14]赵鑫鑫,刘磊,张箴波.Nanog介导FSH诱导卵巢癌转移和侵袭的研究[J].现代妇产科进展,2015,24(5):321-324.
[15]徐春琳,路晓琳,闫晓楠.PI3K/Akt/NF-κB信号通路在FSH促进卵巢癌细胞增殖与侵袭中的作用[J].中华妇产科杂志,2012,47(2):134-138.