奥沙利铂抗人肝癌细胞株QGY增殖及其机制的研究

目的观察抗肿瘤药物奥沙利铂对人肝癌细胞株QGY体外增殖的影响并探讨其机制,为其能否应用于肝癌的临床治疗提供理论依据.方法采用人肝癌细胞株QGY作为研究对象,MTT法检测不同浓度和作用时间的奥沙利铂对细胞增殖的抑制效应;光镜及透射电镜观察细胞形态和超微结构的改变;流式细胞仪分析细胞周期的分布及凋亡;免疫细胞化学法检测细胞周期蛋白及凋亡相关基因蛋白的表达.结果奥沙利铂对QGY细胞的增殖均有抑制作用而且呈时间和剂量依赖性.光镜观察发现细胞体积缩小,变圆,胞浆浓缩,核固缩深染,呈现凋亡早期的形态学改变.电镜可见凋亡细胞及凋亡小体.流式细胞仪检测到细胞阻滞于细胞周期的S期和G2/M期,G0/G1期细胞减少及凋亡峰.奥沙利铂作用72 h后,免疫细胞化学表明cyclin A、Bax表达增强,突变型P53蛋白(MTP53)、Bcl-2、Myc表达减弱,Fas表达无差异.结论奥沙利铂对QGY肝癌细胞的增殖都有一定的抑制作用,阻滞细胞周期于S期及G2/M期,它通过上调Bax的表达,下调MTP53、Bcl-2、Myc的表达而诱导细胞凋亡,该药可能不是通过Fas途径诱导细胞凋亡.

Mechanism of supression on proliferation of human hepatoma cell line QGY by oxaliplatin

Objective To observe the effects of oxaliplatin on proliferation in human hepatoma cell lines QGY in vitro and investigate the mechanism. To provide the theory foundation whether it can be used for the chemotherapy of hepatocellular carcinoma. Methods The inhibition of proliferation in QGY cell was estimated by MTT-test. Morphologic changes were observed under light microscope and electronic microscope. Distribution of cell cycle and apoptosis was analyzed using flow cytometry. The expression of cell cycle protein and apoptosis-associated gene protein was detected with immunohistochemical technique. Results Oxaliplatin could inhibit the proliferation of QGY cells and the inhibition depended on the exposure time and dose. The cells showed morphologic changes at the early stage of apoptosis under the light microscope: the shrunk and round cells, condensed cytoplasma and pycnosis nucleus. Apoptotic cells and apoptotic body could be found under the transmission electronic microscope. The analysis of cell cycle indicated that oxaliplatin blocked cells at S and G_2/M phases and the cells of G_0/G_1 phase reduced. When treated with oxaliplatin for 72 h, the expression of cyclin A and Bax were up-regulated, mutant type P53, Bcl-2 and Myc down-regulated, and Fas was not changed. Conclusion Oxaliplatin could inhibit proliferation of the hepatoma cell lines. Cell cycle blocked at S and G_2/M phase. The apoptosis were related to the up-regulation of Bax and down-regulation of mutant type P53, Bcl-2 and Myc. It could not induce apoptosis through the Fas approach.