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目的观察靶向封闭再生基因Ⅰα(REGⅠα)对胰腺癌细胞株SW1990增殖行为的影响。方法设计并体外合成靶向REGⅠα的小干扰RNA(siRNA),将其转染到SW1990细胞中,半定量RT-PCR、蛋白质印迹法检测转染前后REGⅠα基因表达的变化。采用MTT法检测转染前后细胞增殖能力的改变,流式细胞技术检测细胞周期的变化。结果 REGⅠαsiRNA转染胰腺癌细胞SW1990后,明显抑制REGⅠα基因表达,以200nmol/LREGⅠαsiRNA对REGⅠα基因表达抑制效果最佳。以该浓度的REGⅠαsiRNA转染SW1990细胞后,SW1990细胞在siRNA转染后48hREGⅠαmRNA表达抑制率最高,达82%;干扰后SW1990中REGⅠα蛋白表达也明显下调。抑制SW1990细胞中REGⅠα的表达后,SW1990细胞的增殖能力明显下降,与阴性对照组(转染无关序列的siRNA)及空白对照组(无任何处理)相比,差异有统计学意义(P<0.05)。转染细胞的周期分布有明显改变:G1期细胞增加,S期细胞减少。结论靶向封闭REGⅠα基因能明显抑制胰腺癌细胞SW1990在体外的增殖能力,为探索胰腺癌基因治疗提供新的策略。
Objective To investigate the effect of targeted gene REG1α on the proliferation of pancreatic cancer cell line SW1990. Methods siRNA targeting REGⅠα was designed and synthesized in vitro. The siRNA was transfected into SW1990 cells. The expression of REGⅠα gene was detected by semi-quantitative RT-PCR and Western blotting. MTT assay was used to detect the changes of cell proliferation before and after transfection, and the changes of cell cycle were detected by flow cytometry. Results REGⅠα siRNA transfected into pancreatic cancer cell line SW1990 significantly inhibited the expression of REGⅠα gene and the best inhibitory effect of REGⅠα gene expression with 200nmol / LREGⅠα siRNA. After transfected SW1990 cells with this concentration of REGⅠα siRNA, SW1990 cells had the highest inhibitory rate of 82% at 48h after siRNA transfection, and the expression of REGⅠα protein was also significantly down-regulated in SW1990 after SW1990 treatment. The inhibitory effect of REGⅠα expression in SW1990 cells was significantly decreased compared with that of negative control group (transfected with siRNA unrelated sequence) and blank control group (without any treatment) (P <0.05 ). The cycle distribution of transfected cells changed significantly: cells in G1 phase increased and S phase cells decreased. CONCLUSION: Targeting of REGⅠα gene can significantly inhibit the proliferation of pancreatic cancer cell line SW1990 in vitro and provide a new strategy for gene therapy of pancreatic cancer.