奥沙利铂对人宫颈癌HeLa细胞株体外作用.docVIP

奥沙利铂对人宫颈癌HeLa细胞株体外作用 　【摘要】 【目的】 研究奥沙利铂对人宫颈癌HeLa 细胞株的生长抑制作用及对Caspase-3活性的影响。【方法】 用人宫颈癌HeLa 细胞株经奥沙利铂处理后，MTT法测定细胞生长抑制率,透射电镜观察细胞形态学变化, 流式细胞仪和原位末端标记技术法（TUNEL）检测细胞凋亡率，比色法检测Caspase-3 活性变化，,并利用AC-DEVD-CHO进行Caspase-3抑制试验。【结果】 奥沙利铂 (4～64 μmol/L)在体外能以时间和剂量依赖的方式对宫颈癌HeLa 细胞产生细胞毒作用并诱导其凋亡，奥沙利铂作用细胞株48 h后,细胞呈现典型的凋亡细胞的形态特征；并能增加HeLa 细胞的 Caspase-3 活性，AC-DEVD-乙醛(AC-DEVD-CHO)可有效抑制奥沙利铂诱导的HeLa 细胞凋亡。【结论】 奥沙利铂在体外能抑制人宫颈癌HeLa 细胞的增殖,诱导细胞凋亡， Caspase-3活化参与了奥沙利铂诱导HeLa 细胞的凋亡过程。 【关键词】 宫颈癌； 奥沙利铂； 生长抑制； Caspase-3 Abstract：【Objective】 To study the effect of oxaliplatin on growth inhibition and caspase-3 activity on human cervical cancer HeLa cells. 【Methods】 Human cervical carcinoma HeLa cells were treated with oxaliplatin. Cell growth rate was determined with MTT assay; cell morphologic changes were examined under electron microscopy. Flow cytometry and TUNEL was used to examine cell apoptosis rate. Activities of caspase-3 were determined by chromometry, and its inhibition was test by AC-DEVD-CHO. 【Results】 Oxaliplatin (4-64 μmol/L) induced cytotoxicity and apoptosis to human cervical carcinoma HeLa cells in vitro is time-dependent and dose-dependent. The typical apoptotic morphological features appeared in the cells treated with oxaliplatin for 48 hours; Caspase-3 activity was up-regulated in oxaliplatin-induced apoptotic cells. The oxaliplatin induced HeLa cell apoptosis was efficiently inhibited by AC-DEVD-CHO. 【Conclusion】 Oxaliplatin can inhibit HeLa cell growth of cervical carcinoma and induce apoptosis in vitro. Caspase3 activation may be involved in oxaliplatin-induced apoptosis of HeLa cells. Key words：cervical carcinoma; oxaliplatin; cell growth; caspase-3 [J SUN Yat-sen Univ(Med Sci), 2007, 28(5):485-489] 奥沙利铂(oxaliplatin, L-OHP))是第3代铂类化合物,其化学名为左旋反式二氨环己烷草酸铂。因为与顺铂作用机制有重要区别[1], 对各种顺铂耐药的肿瘤细胞株无交叉耐药,而且该药无明显肾毒性,耳毒性极微,血液学毒性较轻,神经系统毒性主要为剂量依赖性的自限性外周神经病变[2,3]，故近年来不但用于对顺铂耐药的多种人类癌症的二线治疗[4,5]，而且临床上已经有代替顺铂作为一线治疗的使用[6,7]。体内外试验及临床试验已证实了奥沙利铂对晚期大肠癌、胃癌、胰腺癌、卵巢癌、非小细胞肺癌等有较好疗效[8]，但有关奥沙利铂对人宫颈癌的作用及其机制的研究,国内外报道甚少