多柔比星对肝癌细胞GATA4的表达及细胞凋亡的影响  

Effect of doxorubicin on expression of GATA4 and apoptosis of hepatocarcinoma cells

作　　者：任传路[1] 沈红石[2] 王静[3] 刘厚奇[3] 

机构地区：[1]中国人民解放军第100医院检验科,苏州215007 [2]中国人民解放军第100医院血液科,苏州215007 [3]第二军医大学基础医学部组织胚胎学教研室,上海200433

出　　处：《药学服务与研究》2008年第5期336-339,共4页Pharmaceutical Care and Research

基　　金：国家自然科学基金(No.90208026)

摘　　要：目的:通过观察多柔比星(别名阿霉素)对肝癌细胞GATA4表达和细胞凋亡的影响,探讨多柔比星抑制肝癌细胞增殖的可能机制。方法:检测不同p53状态肝癌细胞中GATA4的表达。以300 nmol/L多柔比星加入高表达GATA4的小鼠Hepa1-6肝癌细胞培养液中。碘化丙啶染色,以流式细胞术观察用药后不同时间点(0、6、12、18、24、30 h)凋亡细胞比例,MTT法观察细胞生长,半定量RT-PCR比较不同时间点GATA4、bax、bcl-XL、bcl-2 mRNA的表达,Western blot检测GATA4和细胞增殖核抗原(PCNA)的表达。结果:不同p53状态肝癌细胞中均有GATA4的高表达。分别与0 h或6 h相比,多柔比星处理后12、18、243、0 h的肝癌细胞凋亡率显著增加(流式细胞术检测),且肝癌细胞增殖显著减少(MTT法)。0、6、12、18、24和30 h GATA4 mRNA和GATA4蛋白质的表达呈递减趋势,Bcl-XLmRNA、PCNA蛋白质和bcl-2 mRNA/bax mRNA比值在6 h后也相继递减。结论:GATA4和p53在肝癌细胞中的表达可能不相关。多柔比星可抑制GATA4的表达,而Bcl-XLmRNAb、cl-2 mRNA、bcl-2/bax和PCNA表达的减少,以及凋亡细胞的增多则相对滞后。多柔比星可能通过抑制GATA4的表达来抑制肝癌细胞增殖或诱导肝癌细胞凋亡。Objective： To explore the possible mechanism of doxorubiein （Dox） in inhibiting the proliferation of hepatocar cinoma cells through detecting the influence of Dox on expression of GATA4 and apoptosis in hepatocarcinoma cells. Methods： Expression of GATA4 was detected in hepatocarcinoma cell lines with various p53 activities. Dox （300 nmol/L） was added into the culture medium of Hepal-6 highly expressing GATA4. The ratios of apoptotic cells at different time points after addition of Dox（0, 6, 12, 18, 24, 30 h） were obtained with propidium iodide staining by flow cytometry（FCM）. MTT method was used to observe the cell proliferation. The expression of GATA4, bax, bcl-2 and Bcl-XL mRNA was compared at different time points by semi-quantitative RT PCR. Furthermore, the expression of GATA4 and proliferating cell nuclear antigen （PCNA） was detected by Western blot method. Results： High expression of GATA4 was detected in hepatocarcinoma cell lines with different p53 activities. The result of FCM showed a significant increase in apoptotic ratio of hepatocarcinoma cells treated with Dox at the time points of 12, 18, 24 and 30 h compared with that at 0 h or 6 h, and the result of MTT indicated decreased cell proliferation. The expression of GATA4 mRNA and GATA4 protein presented obvious tendency of degression from 0 h to 30 h, and sequent degression of Bcl-XL mRNA, PCNA protein and ratio of bcl-2 mRNA/bax mRNA appeared 6 h later. Conclusion： There seems to be no obvious correlation between GATA4 and p53. Dox can inhibit the expression of GATA4 while increase in apoptosis and decrease in Bcl-XL mRNA, PCNA protein and the ratio of bcl 2 mRNA and bcl-2 bax mRNA relatively lagged behind. It is implied that Dox might inhibit the proliferation of hepatocarcinoma cells and induce cell apoptosis by depletion of GATA4.

关 键 词：多柔比星 肝肿瘤 细胞凋亡 GATA4 

分 类 号：R965[医药卫生—药理学] R979.14[医药卫生—药学]