顺铂诱导宫颈癌SiHa细胞周期阻滞及凋亡的研究  

Effect of cisplatin on cell cycle arrest and apoptosis of human cervical cancer SiHa cells

作　　者：柳友清[1] 邢辉[1] 韩晓兵[1] 石小燕[1] 梁缝奇[1] 陈刚[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,湖北武汉430030

出　　处：《中华肿瘤防治杂志》2006年第4期248-251,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家973重点基础研究发展规划项目(2002C-B513100)

摘　　要：目的：探讨顺铂在体外对宫颈癌SiHa细胞周期和凋亡的影响及其作用机制。方法：采用MTT法测定顺铂对SiHa细胞增殖的影响；DNA ladder和Hoechst33258检洲药物作用前后的细胞凋亡情况；流式细胞仪检测药物作用前后的周期阻滞和凋亡；Western blot检测E6、p53和p21的表达。结果：顺铂抑制SiHa细胞生长呈时效和量效关系，10mol／L顺铂作用SiHa细胞12、24、36和48h，亚G1峰分别为（3．07±0．15）％、（5．18±0．28）％、（17．73±1．20）％和（32．55±2．74）％，与对照组比较差异均有统计学意义，t值分别为32．80、31．46、29．73和25．99，P值均为0．000；顺铂能使SiHa细胞发生G2～M期阻滞，24h达到最高（66．43±4．62）％，凋亡开始；Hoechst33258实验组可见凋亡细胞；琼脂糖凝肢电泳出现DNA梯形条带；Western blot提示，顺铂作用SiHa细胞后HPV E6的蛋白水平表达逐渐降低，p53和p21蛋白水平表达逐步升高。结论：顺铂随着药物浓度和作用时间的增加对SiHa细胞的体外抑制效应增加，顺铂通过引起SiHa细胞G2～M周期阻滞及抑制HPV E6蛋白的表达，恢复p53功能，启动凋亡，起到杀伤肿瘤作用。OBJECTIVE： To study cell cycle arrest and apoptosis by cisplatin on cervical cancer cell line SiHa and its mechanism in vitro. METHODS： Cell growth inhibition of cisplatin on SiHa was analyzed by MTT assay. Cell apoptosis were detected by DNA cytofluorometry, Hoechst33258 staining and DNA electrophoresis when treated with cisplatin. Cell cycle arrest was analyzed by cytometry. The protein expressions of E6, p53 and p21 were studied by Western blot. RESULTS： Cisplatin inhibited the proliferation of SiHa cells in a time and dose-dependant manner. After 12, 24, 36, 48 h treated with 10mol/L Cisplatin, Sub-G1 peaks detected by flow cytometry were （3. 07±0. 15）%, （5. 18±0. 28）%, （17. 73±1.20）% and （32. 55±2.74）%, respectively. The percentages of sub-G1 of the experimental group were significantly higher than those of the control （t = 32. 80, 31.46, 29. 73 and 25. 99, P = 0. 000）% Hoechst33258 staining, DNA ladder and cytomytry showed apoptosis induced by cisplatin. Cisplatin caused G2-M cell cycle arrest. Western blot showed cisplatin decreased expression of HPV E6 and increased expressions of p53 and p21 proteins. CONCLUSION： Cisplatin can induce cycle arrest and restore the function of p53 to induce apoptosis in SiHa cells to kill the tumor cell.

关 键 词：宫颈肿瘤/病理学 顺铂/药理学 细胞凋亡 病毒蛋白质类 蛋白质P53 蛋白质P21 流式细胞术 

分 类 号：R737.33[医药卫生—肿瘤]