丹参酮IIA抑制前列腺癌细胞株生长和诱导凋亡的试验研究
|
摘要
目的
通过体外细胞培养的方法,证实丹参酮ⅡA(TanⅡA)体外抑制人前列腺癌细胞株PC3和DU145生长及诱导凋亡的作用,并进一步分析其对细胞周期和凋亡相关基因蛋白表达的影响。探讨丹参酮ⅡA在PC3和DU145中可能的作用机制,最终希望为晚期前列腺癌的临床治疗提供新选择。
方法
体外培养人前列腺癌细胞株PC3和DU145,分别用不同浓度的丹参酮ⅡA(2.5,5.0,10.0,20.0mg.L~(-1))处理细胞24,48,72小时后进行如下指标检测:
1.光镜,电镜观察细胞形态改变;
2.细胞计数、细胞毒试验(CCK-8法)、集落形成试验检测细胞生长和增殖的抑制情况;
3.琼脂糖凝胶电泳分析细胞DNA“梯状”断裂;
4.流式细胞术PI染色检测用药前后细胞周期变化;
5.Western-blot检测细胞凋亡相关基因蛋白p53,p21,bcl-2,Caspase3的表达改变,探讨药物作用机制。
结果
1.细胞形态观察:光镜下可见对照组细胞贴壁生长,类圆形或梭形,体积较大,排列紧密,边缘光滑;经10.0mg.L~(-1)丹参酮ⅡA处理后,细胞排列稀疏,细胞体积缩小,胞膜皱缩,成小圆形或不规则形态,并见到较多细胞漂浮于培养基中。随着作用时间延长(24h-48h-72h),其形态改变的程度逐渐升高;透射电镜可见TanⅡA组细胞发生了凋亡的特征性改变如核固缩、碎裂,凋亡小体形成。
2.细胞毒试验:各浓度的丹参酮ⅡA(2.5,5.0,10.0,20.0mg.L~(-1))对PC3和DU145的生长均有抑制作用,明显强于对照组,其差别有统计学意义(P＜0.05)。此外,试验发现,随作用时间延长和丹参酮ⅡA浓度增加,其生长抑制作用逐渐增强。经双因素方差分析检验,10.1mg.L~(-1)丹参酮ⅡA处理不同时间的细胞存活率之间有统计学差异(P＜0.05),不同浓度丹参酮ⅡA处理72h后的细胞存活率之间也有统计学差异(P＜0.05),提示药物作用有明显的时间、浓度依赖性。
3.集落形成试验:经丹参酮ⅡA处理后的PC3和DU145细胞集落形成率明显降低,与对照组相比有统计学差异(P＜0.05)。并且随药物浓度上升集落形成率逐渐下降。
4.琼脂糖凝胶电泳:经2.5,5.0,10.0 mg.L~(-1)丹参酮ⅡA处理的DU145细胞和5.0,10.0 mg.L~(-1)处理的PC3细胞均发生了DNA“梯状”断裂,而对照组未发生此现象。
5.流式细胞术检测细胞周期:经不同浓度丹参酮ⅡA(5.0,10.0,20.0mg.L~(-1))作用后的PC3和DU145细胞,G_0/G_1期比例显著升高,S期比例明显减少,细胞凋亡率上升。
6.凋亡相关基因蛋白表达检测:Western-blot检测发现,与对照组相比,丹参酮ⅡA可明显上调p21的蛋白表达,下调bcl-2的蛋白表达,并能不同程度上调p53和Caspase3的表达水平。
结论
丹参酮ⅡA在体外具有明显的抑制前列腺癌细胞株PC3和DU145生长,诱导凋亡的作用,该作用有时间、浓度依赖性.丹参酮ⅡA可能的作用机制是使细胞阻滞于G_0/G_1期,明显降低细胞增殖指数,并影响凋亡相关基因蛋白的表达,如上调p53,p21,Caspase3的表达,下调bcl-2表达,从而诱导肿瘤细胞发生凋亡。试验表明,丹参酮ⅡA在前列腺癌的临床治疗方面有潜在价值,值得进一步研究探索。
Objective
To evaluate the effect of tanshinoneⅡA on growth inhibition and apoptosis induction in human prostate cancer cell lines(PC3 and DU145) in vitro.To investigate the cell cycle and apoptosis-related genes protein expression changes of PC3 and DU145 cells treated with tanshinoneⅡA.Explore the possible mechanism of tanshinoneⅡA and provide a gist for the treatment of advanced prostate cancer.
Methods
The PC3 and DU145 cells were cultured in vitro and treated with tanshinoneⅡA in different dose(2.5,5.0,10.0,20.0mg.L~(-1)) and different time(24h,48h,72h) for following measurement.
1.Observation of morphologic changes under the light microscope(LM) and transmission electron microscope(TEM).
2.CCK-8 assay and colony-forming assay were used to investigate the inhibition of cells' growth.
3.Agarose gel electrophoresis was used to examine the DNA fragments.
4.Changes of cell cycle were analyzed with PI staining,flow cytometry(FCM).
5.Western-blot assay was used to detect the apoptosis-related genes p53,p21,bcl-2, Caspase3 protein expression changes.
Results
1.LM and TEM observation:Under light microscope,the PC3 and DU145 cells in control group were large,roundish and serried.After treated by 10.0mg.L~(-1) tanshinoneⅡA,the cells became small,detached or sparse and membranous frothed or wizened.The characteristic morphologic changes of apoptosis such as chromatin condensation and apoptotic body were observed under transmission electron microscope.
2.Cytotoxicity test:In CCK-8 assay,all the tanshinoneⅡA dose levels(2.5,5.0,10.0, 20.0mg.L~(-1)) induced growth inhibition in both of the 2 cell lines.The inhibitory effect became stronger with the passage of time after treatment with tanshinoneⅡA,as well as the increase of doses.The differences between different dose and treatment time groups were statistically significant,which demonstrated the growth and proliferation of PC3 and DU145 cells were inhibited in a dose- and time- dependent manner.
3.Colony-forming assay:The colony-forming rates in both of PC3 and DU145 cells were obviously suppressed by every dose level oftanshinoneⅡA.
4.Agarose gel electrophoresis:DNA "ladder" was presented in 2.5,5.0 and 10.0 mg.L~(-1) tanshinoneⅡA groups in DU145 cells,and in 5.0,10.0 mg.L~(-1) groups in PC3 cells,but not observed in control groups.
5.Cell cycle analysis:Compared with control groups,the cells treated with tanshinoneⅡA were arrested in G_0/G_1 phase and decreased in S phase.The apoptosis index was increased in flow cytometry.
6.Western-blot assay:Compared with control groups,the protein expression of apoptosis-related genes p53,p21,Caspase3 in treated cells were up-regulated and bcl-2 down-regulated.
Conclusion
The findings of the study suggested that tanshinoneⅡA could inhibit the growth and induce apoptosis in PC3 and DU145 cells in vitro.The possible mechanisms of tanshinoneⅡA were arresting the cell cycle in G_0/G_1 phase,up-regulating p53,p21, Caspase3 expression of protein and down-regulating bcl-2 expression.TanshinoneⅡA could be a potential effective candidate for the clinical treatment of advanced prostate cancer patients.
通过体外细胞培养的方法,证实丹参酮ⅡA(TanⅡA)体外抑制人前列腺癌细胞株PC3和DU145生长及诱导凋亡的作用,并进一步分析其对细胞周期和凋亡相关基因蛋白表达的影响。探讨丹参酮ⅡA在PC3和DU145中可能的作用机制,最终希望为晚期前列腺癌的临床治疗提供新选择。
方法
体外培养人前列腺癌细胞株PC3和DU145,分别用不同浓度的丹参酮ⅡA(2.5,5.0,10.0,20.0mg.L~(-1))处理细胞24,48,72小时后进行如下指标检测:
1.光镜,电镜观察细胞形态改变;
2.细胞计数、细胞毒试验(CCK-8法)、集落形成试验检测细胞生长和增殖的抑制情况;
3.琼脂糖凝胶电泳分析细胞DNA“梯状”断裂;
4.流式细胞术PI染色检测用药前后细胞周期变化;
5.Western-blot检测细胞凋亡相关基因蛋白p53,p21,bcl-2,Caspase3的表达改变,探讨药物作用机制。
结果
1.细胞形态观察:光镜下可见对照组细胞贴壁生长,类圆形或梭形,体积较大,排列紧密,边缘光滑;经10.0mg.L~(-1)丹参酮ⅡA处理后,细胞排列稀疏,细胞体积缩小,胞膜皱缩,成小圆形或不规则形态,并见到较多细胞漂浮于培养基中。随着作用时间延长(24h-48h-72h),其形态改变的程度逐渐升高;透射电镜可见TanⅡA组细胞发生了凋亡的特征性改变如核固缩、碎裂,凋亡小体形成。
2.细胞毒试验:各浓度的丹参酮ⅡA(2.5,5.0,10.0,20.0mg.L~(-1))对PC3和DU145的生长均有抑制作用,明显强于对照组,其差别有统计学意义(P＜0.05)。此外,试验发现,随作用时间延长和丹参酮ⅡA浓度增加,其生长抑制作用逐渐增强。经双因素方差分析检验,10.1mg.L~(-1)丹参酮ⅡA处理不同时间的细胞存活率之间有统计学差异(P＜0.05),不同浓度丹参酮ⅡA处理72h后的细胞存活率之间也有统计学差异(P＜0.05),提示药物作用有明显的时间、浓度依赖性。
3.集落形成试验:经丹参酮ⅡA处理后的PC3和DU145细胞集落形成率明显降低,与对照组相比有统计学差异(P＜0.05)。并且随药物浓度上升集落形成率逐渐下降。
4.琼脂糖凝胶电泳:经2.5,5.0,10.0 mg.L~(-1)丹参酮ⅡA处理的DU145细胞和5.0,10.0 mg.L~(-1)处理的PC3细胞均发生了DNA“梯状”断裂,而对照组未发生此现象。
5.流式细胞术检测细胞周期:经不同浓度丹参酮ⅡA(5.0,10.0,20.0mg.L~(-1))作用后的PC3和DU145细胞,G_0/G_1期比例显著升高,S期比例明显减少,细胞凋亡率上升。
6.凋亡相关基因蛋白表达检测:Western-blot检测发现,与对照组相比,丹参酮ⅡA可明显上调p21的蛋白表达,下调bcl-2的蛋白表达,并能不同程度上调p53和Caspase3的表达水平。
结论
丹参酮ⅡA在体外具有明显的抑制前列腺癌细胞株PC3和DU145生长,诱导凋亡的作用,该作用有时间、浓度依赖性.丹参酮ⅡA可能的作用机制是使细胞阻滞于G_0/G_1期,明显降低细胞增殖指数,并影响凋亡相关基因蛋白的表达,如上调p53,p21,Caspase3的表达,下调bcl-2表达,从而诱导肿瘤细胞发生凋亡。试验表明,丹参酮ⅡA在前列腺癌的临床治疗方面有潜在价值,值得进一步研究探索。
Objective
To evaluate the effect of tanshinoneⅡA on growth inhibition and apoptosis induction in human prostate cancer cell lines(PC3 and DU145) in vitro.To investigate the cell cycle and apoptosis-related genes protein expression changes of PC3 and DU145 cells treated with tanshinoneⅡA.Explore the possible mechanism of tanshinoneⅡA and provide a gist for the treatment of advanced prostate cancer.
Methods
The PC3 and DU145 cells were cultured in vitro and treated with tanshinoneⅡA in different dose(2.5,5.0,10.0,20.0mg.L~(-1)) and different time(24h,48h,72h) for following measurement.
1.Observation of morphologic changes under the light microscope(LM) and transmission electron microscope(TEM).
2.CCK-8 assay and colony-forming assay were used to investigate the inhibition of cells' growth.
3.Agarose gel electrophoresis was used to examine the DNA fragments.
4.Changes of cell cycle were analyzed with PI staining,flow cytometry(FCM).
5.Western-blot assay was used to detect the apoptosis-related genes p53,p21,bcl-2, Caspase3 protein expression changes.
Results
1.LM and TEM observation:Under light microscope,the PC3 and DU145 cells in control group were large,roundish and serried.After treated by 10.0mg.L~(-1) tanshinoneⅡA,the cells became small,detached or sparse and membranous frothed or wizened.The characteristic morphologic changes of apoptosis such as chromatin condensation and apoptotic body were observed under transmission electron microscope.
2.Cytotoxicity test:In CCK-8 assay,all the tanshinoneⅡA dose levels(2.5,5.0,10.0, 20.0mg.L~(-1)) induced growth inhibition in both of the 2 cell lines.The inhibitory effect became stronger with the passage of time after treatment with tanshinoneⅡA,as well as the increase of doses.The differences between different dose and treatment time groups were statistically significant,which demonstrated the growth and proliferation of PC3 and DU145 cells were inhibited in a dose- and time- dependent manner.
3.Colony-forming assay:The colony-forming rates in both of PC3 and DU145 cells were obviously suppressed by every dose level oftanshinoneⅡA.
4.Agarose gel electrophoresis:DNA "ladder" was presented in 2.5,5.0 and 10.0 mg.L~(-1) tanshinoneⅡA groups in DU145 cells,and in 5.0,10.0 mg.L~(-1) groups in PC3 cells,but not observed in control groups.
5.Cell cycle analysis:Compared with control groups,the cells treated with tanshinoneⅡA were arrested in G_0/G_1 phase and decreased in S phase.The apoptosis index was increased in flow cytometry.
6.Western-blot assay:Compared with control groups,the protein expression of apoptosis-related genes p53,p21,Caspase3 in treated cells were up-regulated and bcl-2 down-regulated.
Conclusion
The findings of the study suggested that tanshinoneⅡA could inhibit the growth and induce apoptosis in PC3 and DU145 cells in vitro.The possible mechanisms of tanshinoneⅡA were arresting the cell cycle in G_0/G_1 phase,up-regulating p53,p21, Caspase3 expression of protein and down-regulating bcl-2 expression.TanshinoneⅡA could be a potential effective candidate for the clinical treatment of advanced prostate cancer patients.
引文
1.Luo HW,Sun XR,Niwa Masatake.Diterpenoids from Salvia paramiltionrrhiza[J].Heterocycles,1994,38(11):2473-2479.
2.房其年,张佩玲,徐宗沛。丹参抗菌有效成分的研究[J]。化学学报,1976,34(3):197-209。
3.杨艺。丹参酮药理作用近识[J]。湖北中医杂志,1999,21(6):284-286。
4.梁勇,羊裔明,袁淑兰。丹参酮药理作用及临床应用研究进展[J]。中草药,2000,31(4):304-306。
5.LeeD S,LeeS H,KwonG S,et al.In hibition of DNA Topoisomerase I by Dihydrotanshinone I,component of a medicinal herb Salvia miltiorrhiza Bunge[J].BiosciBiotechem iochem,1999,63(8):1370-1373.
6.袁淑兰。丹参酮在制备治疗肿瘤药物中的应用[J]。辽宁医药,2001,16(3):36-39。
7.王修杰,袁淑兰,黄韧敏等。Brdu和PCNA标记观察丹参酮对癌细胞增殖的影响[J]。华西医科大学学报,1996,27(4):388-391。
8.王修杰,袁淑兰,王朝俊等。丹参酮抗小鼠肝癌作用和机理的初步研究。中华肿瘤杂志,1996,18(6):412-414。
9.Wang XJ,Yuan SL,Wang CJ et al.A preliminary study of the anticancer effect of tanshinone on hepatic cancer and its mechanism of action in mice[J].China Cancer Res,1998,10(2):100-102.
10.Yoon Y,Kim YO,Jeon WK et al.Tanshinone II A isolated from Salvia miltiorrhiza BUNGE induced apoptosis in HL60 human premyelocytic leukemia cell line[J].J Ethnohparmacology,1999,68(1-3):121-127.
11.Sung HJ,Choi SM,Yoon Y et al.Tanshinone II A,an ingredient of Salvia miltiorrhiza BUNGE,induces apoptosis in human leukemia cell lines through the activation of caspase-3[J].Experimental & Molecular Medicine,1999,31(4):174-178.
12.张培彤,裴迎霞,祁鑫等。不同药物对人肺巨细胞癌PGCL3细胞表面表达的 血小板免疫相关抗原的影响[J]。中草药,1999,30(5):352-355。
13.敬静,郑鸿,王静等。丹参酮IIA对人ER阴性乳腺癌细胞的生长抑制和多耐药逆转作用。四川大学学报(医学版),2007,38(3):391-395。
14.司徒镇强,吴军正。细胞培养(修订版)[M]。西安:世界图书出版公司,2004:298-303。
15.陈竹。丹参酮ⅡA对Hela细胞的细胞增殖和细胞周期的影响[D]。太原:山西医科大学,2006:
16.Peter ME,Heufelder AE,Hengartner MO.Advances in apoptosis research.Proc Nail Acad Sci USA,1997,94:12736-12737.
17.Bergamaschi G,Rosti V,Danova M,Lucotti C,Cazzola M.Apoptosis:Biological and clinical aspects.Haematologica,1994,79:86-90.
18.Marx J.Cell death studies yield cancer clues.Science,1998,259(5096):760-761.
19.Tsuda H,Sata M,ljuuin H,Kumabe T,Uchida M,Ogou Y,Akagi Y,Shirouzu K,Hara H,Nakashima Y.A novel strategy for remission induction and maintenance in cancer therapy,Oncol Rep,2002,9(1):65-68.
20.Altieri DC.Survivin,versatile modulation of cell division and apoptosis in cancer.Oncogene,2003,22:8581-8589.
21.Kerr JF,Winterford CM,Harmon BV,Apoptosis:its significance in cancer and cancer therapy.Cancer,1997,73(8):2013-2026.
22.Zhu L,Harlow E,Dynlacht BD.P~(107) uses a p21CIPl-related domain to bind cyclin/cdk2 and regulate interactions with E2F[J].Genes Dev,1995,9(14):1740-1752
23.谭立军,沈忠英,汤雪明。细胞周期调控与肿瘤.肿瘤,1997,17(5):295-297。
24.El-Deiry WS,Tokino T,Velculescu VE et al.A potential mediator of p53 tumor suppression.Cell,1993,75(4):817-825.
25.EI-Deiry WS,Harper JW,O'Connor PM et al.WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis.Cancer Res,1994,54(5):1169-1174.
26.Hunter T,Pines J.Cyclins and cancer Ⅱ:cyclin D and CDK inhibitors come of age.Cell,1994,79(4):573-582.
1.Ryu SY,Lee CO,choi SU et al.In Vitro cytotxicity of tanshinones from Salvia miltiorrhiza[J].Plant medica,1997,63(4):339-342.
2.袁淑兰,羊裔明,黄光琦。应用中药提取物丹参酮治疗肿瘤[P]。中国专利:1264580A,2002-08-30。
3.Mosaddik MA.In vitro cytotoxicity of tanshinones isolated from Salvia miltiorrhiza Bunge against P388 lymphocytic leukemia cells[J].Phytomedicine,2003,10(8):682-685.
4.陈良良,吴良村,李庆霞等。丹参注射液对肿瘤浸润淋巴细胞的体外扩增及抗肿瘤作用[J]。浙江中医学院学报,2002,26(3):49-52。
5.梁勇,羊裔明,袁淑兰等。丹参酮ⅡA诱导急性早幼粒细胞白血病分化及其分子机制[J]。中华血液杂志,2000,21(1):23-26。
6.黄光琦,袁淑兰,周宏远等。丹参诱导人宫颈癌肛180细胞的分析[J]。中华药理学和毒理学杂志,1996,10(4):285。
7.吴俣,羊裔明,孟文彤等。丹参酮对K562细胞株的诱导分化[J]。华西医科大学学报,2002,33(1):80-83。
8.徐俊丽,和水祥,陈静宏等。丹参酮ⅡA对肝癌SMMC-7721细胞COX-2表达的影响[J]。世界华人消化杂志,2006,14(14):1352-1356。
9.唐忠志,唐瑛,付立波。丹参酮ⅡA对人肝癌BEL-7402细胞生长的影响及其机制[J]。第一军医大学学报,2003,23(6):595-601。
10.李庆彦,李春忠,黄智武等。3种丹参制剂对Bel-7402细胞和外周血干细胞生长的影响[J]。实用癌症杂志,2001,16(5):467-468。
11.刘明章,黄贻穗,肖伟琪。丹参酮ⅡA磺酸钠对Lewis癌无促进生长与转移的作用[J]。中国药理学报,1991,12(6):534-537。
12.张培彤,裴迎霞,祁鑫等。不同药物对人肺巨细胞癌PGCL3细胞表面表达的血小板免疫相关抗原的影响[J]。中草药,1999,30(5):353-355。
13.孙婧强,周信达,刘银坤。丹参对肝癌转移复发防治作用的研究[J]。中国中西医结合杂志,1999,19(5):292-295。
14.刘鲁明,陈震,陈培丰。对活血化瘀中药治疗恶性肿瘤的思考[J]。中医杂志,2007,48(9):776-779。
15.夏荣龙,陈孝平。丹参在大鼠恶性梗阻性黄疸时对肿瘤细胞凋亡的作用[J]。第四军医大学学报,2006,27(19):1802-1804。
16.Xiujie Wang,Yuquan Wei,Shulan Yuan et al.Potiential anticancer activity of tanshinone ⅡA against human breast cancer.Int.J.Cancer,2005,116:799 - 807.
17.Bai S,Zhao Y,Zhang X,Li H,Jing J,Xie C,Chen G Establishment of a heterologous graft model for human breast infiltrating duct carcinoma in nude mice.Hua Xi Yi Ke Da Xue Xue Bao,1999,30:220 - 221.
18.Chen D,Zhao Y,Bai S,Shi Z,Zhang J.Effect ofginsenoside Rg3 on the progression of orthotopically xenotransplanted human breast cancer in nude mice and its mechanism.Sichuan Da Xue Xue Bao Yi Xue Bao,2003,34:546 - 548.
19.Liu Chen H,Jiang Y.Protective effect of tanshinone ⅡA on acute hepatic injury in mice.Zhong Yao Cai,2001,24:588 - 589.
20.Singh AV,Xiao D,Lew KL,Dhir R,Singh SV.Sulforaphane induces caspase-mediated apoptosis in cultured PC-3 human prostate cancer cells and retards growth of PC-3 xenografts in vivo.Carcinogenesis,2004.25:83 - 90.
2.房其年,张佩玲,徐宗沛。丹参抗菌有效成分的研究[J]。化学学报,1976,34(3):197-209。
3.杨艺。丹参酮药理作用近识[J]。湖北中医杂志,1999,21(6):284-286。
4.梁勇,羊裔明,袁淑兰。丹参酮药理作用及临床应用研究进展[J]。中草药,2000,31(4):304-306。
5.LeeD S,LeeS H,KwonG S,et al.In hibition of DNA Topoisomerase I by Dihydrotanshinone I,component of a medicinal herb Salvia miltiorrhiza Bunge[J].BiosciBiotechem iochem,1999,63(8):1370-1373.
6.袁淑兰。丹参酮在制备治疗肿瘤药物中的应用[J]。辽宁医药,2001,16(3):36-39。
7.王修杰,袁淑兰,黄韧敏等。Brdu和PCNA标记观察丹参酮对癌细胞增殖的影响[J]。华西医科大学学报,1996,27(4):388-391。
8.王修杰,袁淑兰,王朝俊等。丹参酮抗小鼠肝癌作用和机理的初步研究。中华肿瘤杂志,1996,18(6):412-414。
9.Wang XJ,Yuan SL,Wang CJ et al.A preliminary study of the anticancer effect of tanshinone on hepatic cancer and its mechanism of action in mice[J].China Cancer Res,1998,10(2):100-102.
10.Yoon Y,Kim YO,Jeon WK et al.Tanshinone II A isolated from Salvia miltiorrhiza BUNGE induced apoptosis in HL60 human premyelocytic leukemia cell line[J].J Ethnohparmacology,1999,68(1-3):121-127.
11.Sung HJ,Choi SM,Yoon Y et al.Tanshinone II A,an ingredient of Salvia miltiorrhiza BUNGE,induces apoptosis in human leukemia cell lines through the activation of caspase-3[J].Experimental & Molecular Medicine,1999,31(4):174-178.
12.张培彤,裴迎霞,祁鑫等。不同药物对人肺巨细胞癌PGCL3细胞表面表达的 血小板免疫相关抗原的影响[J]。中草药,1999,30(5):352-355。
13.敬静,郑鸿,王静等。丹参酮IIA对人ER阴性乳腺癌细胞的生长抑制和多耐药逆转作用。四川大学学报(医学版),2007,38(3):391-395。
14.司徒镇强,吴军正。细胞培养(修订版)[M]。西安:世界图书出版公司,2004:298-303。
15.陈竹。丹参酮ⅡA对Hela细胞的细胞增殖和细胞周期的影响[D]。太原:山西医科大学,2006:
16.Peter ME,Heufelder AE,Hengartner MO.Advances in apoptosis research.Proc Nail Acad Sci USA,1997,94:12736-12737.
17.Bergamaschi G,Rosti V,Danova M,Lucotti C,Cazzola M.Apoptosis:Biological and clinical aspects.Haematologica,1994,79:86-90.
18.Marx J.Cell death studies yield cancer clues.Science,1998,259(5096):760-761.
19.Tsuda H,Sata M,ljuuin H,Kumabe T,Uchida M,Ogou Y,Akagi Y,Shirouzu K,Hara H,Nakashima Y.A novel strategy for remission induction and maintenance in cancer therapy,Oncol Rep,2002,9(1):65-68.
20.Altieri DC.Survivin,versatile modulation of cell division and apoptosis in cancer.Oncogene,2003,22:8581-8589.
21.Kerr JF,Winterford CM,Harmon BV,Apoptosis:its significance in cancer and cancer therapy.Cancer,1997,73(8):2013-2026.
22.Zhu L,Harlow E,Dynlacht BD.P~(107) uses a p21CIPl-related domain to bind cyclin/cdk2 and regulate interactions with E2F[J].Genes Dev,1995,9(14):1740-1752
23.谭立军,沈忠英,汤雪明。细胞周期调控与肿瘤.肿瘤,1997,17(5):295-297。
24.El-Deiry WS,Tokino T,Velculescu VE et al.A potential mediator of p53 tumor suppression.Cell,1993,75(4):817-825.
25.EI-Deiry WS,Harper JW,O'Connor PM et al.WAF1/CIP1 is induced in p53-mediated G1 arrest and apoptosis.Cancer Res,1994,54(5):1169-1174.
26.Hunter T,Pines J.Cyclins and cancer Ⅱ:cyclin D and CDK inhibitors come of age.Cell,1994,79(4):573-582.
1.Ryu SY,Lee CO,choi SU et al.In Vitro cytotxicity of tanshinones from Salvia miltiorrhiza[J].Plant medica,1997,63(4):339-342.
2.袁淑兰,羊裔明,黄光琦。应用中药提取物丹参酮治疗肿瘤[P]。中国专利:1264580A,2002-08-30。
3.Mosaddik MA.In vitro cytotoxicity of tanshinones isolated from Salvia miltiorrhiza Bunge against P388 lymphocytic leukemia cells[J].Phytomedicine,2003,10(8):682-685.
4.陈良良,吴良村,李庆霞等。丹参注射液对肿瘤浸润淋巴细胞的体外扩增及抗肿瘤作用[J]。浙江中医学院学报,2002,26(3):49-52。
5.梁勇,羊裔明,袁淑兰等。丹参酮ⅡA诱导急性早幼粒细胞白血病分化及其分子机制[J]。中华血液杂志,2000,21(1):23-26。
6.黄光琦,袁淑兰,周宏远等。丹参诱导人宫颈癌肛180细胞的分析[J]。中华药理学和毒理学杂志,1996,10(4):285。
7.吴俣,羊裔明,孟文彤等。丹参酮对K562细胞株的诱导分化[J]。华西医科大学学报,2002,33(1):80-83。
8.徐俊丽,和水祥,陈静宏等。丹参酮ⅡA对肝癌SMMC-7721细胞COX-2表达的影响[J]。世界华人消化杂志,2006,14(14):1352-1356。
9.唐忠志,唐瑛,付立波。丹参酮ⅡA对人肝癌BEL-7402细胞生长的影响及其机制[J]。第一军医大学学报,2003,23(6):595-601。
10.李庆彦,李春忠,黄智武等。3种丹参制剂对Bel-7402细胞和外周血干细胞生长的影响[J]。实用癌症杂志,2001,16(5):467-468。
11.刘明章,黄贻穗,肖伟琪。丹参酮ⅡA磺酸钠对Lewis癌无促进生长与转移的作用[J]。中国药理学报,1991,12(6):534-537。
12.张培彤,裴迎霞,祁鑫等。不同药物对人肺巨细胞癌PGCL3细胞表面表达的血小板免疫相关抗原的影响[J]。中草药,1999,30(5):353-355。
13.孙婧强,周信达,刘银坤。丹参对肝癌转移复发防治作用的研究[J]。中国中西医结合杂志,1999,19(5):292-295。
14.刘鲁明,陈震,陈培丰。对活血化瘀中药治疗恶性肿瘤的思考[J]。中医杂志,2007,48(9):776-779。
15.夏荣龙,陈孝平。丹参在大鼠恶性梗阻性黄疸时对肿瘤细胞凋亡的作用[J]。第四军医大学学报,2006,27(19):1802-1804。
16.Xiujie Wang,Yuquan Wei,Shulan Yuan et al.Potiential anticancer activity of tanshinone ⅡA against human breast cancer.Int.J.Cancer,2005,116:799 - 807.
17.Bai S,Zhao Y,Zhang X,Li H,Jing J,Xie C,Chen G Establishment of a heterologous graft model for human breast infiltrating duct carcinoma in nude mice.Hua Xi Yi Ke Da Xue Xue Bao,1999,30:220 - 221.
18.Chen D,Zhao Y,Bai S,Shi Z,Zhang J.Effect ofginsenoside Rg3 on the progression of orthotopically xenotransplanted human breast cancer in nude mice and its mechanism.Sichuan Da Xue Xue Bao Yi Xue Bao,2003,34:546 - 548.
19.Liu Chen H,Jiang Y.Protective effect of tanshinone ⅡA on acute hepatic injury in mice.Zhong Yao Cai,2001,24:588 - 589.
20.Singh AV,Xiao D,Lew KL,Dhir R,Singh SV.Sulforaphane induces caspase-mediated apoptosis in cultured PC-3 human prostate cancer cells and retards growth of PC-3 xenografts in vivo.Carcinogenesis,2004.25:83 - 90.