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目的:探讨Pokemon特异性小干扰RNA对肺癌细胞A549和食管癌细胞EC109增殖的影响。方法:瞬时转染Pokemon特异性小干扰RNA至肺癌细胞A549和食管癌细胞EC109,RT-PCR、Western Blot技术检测转染后细胞中Pokemon的mRNA和蛋白表达水平,检测细胞的增殖及细胞周期变化。结果:与空白组和阴性对照组相比,瞬时转染Pokemon小干扰RNA后,肺癌细胞A549和食管癌细胞EC109中Pokemon的mRNA水平均下降至25%~35%,蛋白水平亦明显下降。细胞增殖能力在培养24,48,72 h均显著降低(P<0.05)。细胞周期分析显示转染Pokemon小干扰RNA后S期的比例显著高于siRNA阴性对照组(A549细胞:55.7%±2.5%vs 42.7%±0.6%,P<0.01;EC109细胞:67.7%±2.5%vs 52.0%±2.0%,P<0.01)。G1期的比例显著低于siRNA阴性对照组(A549细胞:33.0%±2.0%vs 45.3%±1.5%,P<0.01;EC109细胞:30.7%±1.2%vs 44.0%±1.7%,P<0.01)。两种细胞均阻滞于S期。结论:Pokemon小干扰RNA可抑制肺癌细胞A549和食管癌细胞EC109的增殖。
Objective: To investigate the effect of Pokemon-specific small interfering RNA on the proliferation of lung cancer cell line A549 and esophageal cancer cell line EC109. METHODS: Pokemon-specific small interfering RNA was transiently transfected into lung cancer cell line A549 and esophageal cancer cell line EC109. The expression of Pokemon mRNA and protein in transfected cells was detected by RT-PCR and Western Blot. The proliferation and cell cycle changes were detected. . RESULTS: Compared with the blank group and the negative control group, the Pokemon mRNA levels in lung cancer cells A549 and esophageal cancer EC109 decreased to 25% to 35% after transient transfection of Pokemon siRNA, and the protein levels also decreased significantly. The cell proliferation was significantly decreased at 24, 48, and 72 h (P<0.05). Cell cycle analysis showed that the proportion of S phase after transfection of Pokemon small interfering RNA was significantly higher than that of siRNA negative control group (A549 cells: 55.7%±2.5% vs 42.7%±0.6%, P<0.01; EC109 cells: 67.7%±2.5% Vs 52.0% ± 2.0%, P < 0.01). The proportion of G1 phase was significantly lower than that of siRNA negative control group (A549 cells: 33.0%±2.0% vs 45.3%±1.5%, P<0.01; EC109 cells: 30.7%±1.2% vs 44.0%±1.7%, P<0.01) . Both cells arrested in the S phase. Conclusion: Pokemon siRNA can inhibit the proliferation of lung cancer cells A549 and esophageal cancer EC109.