Observation on sensitivity of human gastric cancer SGC-7901 cells to cisplatin by RNAi techniques
Observation on sensitivity of human gastric cancer SGC-7901 cells to cisplatin by RNAi techniques
复制标题
RNAi技术观察人胃癌SGC-7901细胞对顺铂的敏感性
DOI：
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发表时间：
2009
期刊：
影响因子：
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通讯作者：
师建国
中科院分区：
文献类型：
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作者：
臧丹丹;杨世明;闫庆国;崔颖;师建国
目的：研究RNAi技术下调Rb蛋白表达是否能增强人胃癌SGC-7901细胞对顺铂的敏感性。方法：将靶向Rb的siRNA和阴性对照siRNA分别导入SGC-7901细胞。RT-PCR和Western印迹检测基因敲除效率。用流式细胞仪分析细胞周期分布。用10%胎牛血清或无血清培养细胞，用顺铂处理不同时间。用活化的caspase-3分析细胞的凋亡。用四甲基偶氮唑盐比色法检测细胞存活率。结果：靶向Rb的siRNA显著降低了细胞的蛋白质和RNA水平，使细胞从G1期解离。当细胞在10%的血清中培养时，用20μM顺铂处理这些细胞，与阴性对照siRNAs细胞相比，细胞凋亡率增加了约1.8%。相反，无血清培养的细胞在Rb抑制组和对照组之间无显著差异。结论：RNAi技术下调Rb基因表达，通过促进细胞进入G1期，增强了胃癌细胞对顺铂的化疗敏感性。
Objective: In our study,we investigated whether down-regulating Rb protein by RNAi techniques can enhance the sensitivity of human gastric cancer SGC-7901 cells to cisplatin. Methods: SiRNA targeting Rb or the negative control siRNA was transfected into SGC-7901 cells. Efficiency of knockdown was tested by RT-PCR and Western blot. Cell cycle distribution was analyzed by FACS. Cells with siRNA transfection were cultured in 10% FBS or serum free medium and were treated with cisplatin for different periods. Apoptosis was analyzed by activated caspase-3. Cell survival was analyzed by MTT assay. Results: An siRNA targeting Rb significantly decreased the protein and RNA level, which released the cells from G1 phase. Treatment of these cells with 20 μM cisplatin increased apoptotic cell death approximately 1.8-fold compared to cells transfected with negative control siRNA when cells were cultured in 10% serum. In contrast, cells cultured in the serum free medium have no significant differences between Rb inhibition and the control. Conclusions: Our data demonstrate that down-regulation of Rb using RNAi techniques enhances the chemosensitivity of gastric cancer cells to cisplatin through facilitating cells enter the G1 phase.