论文部分内容阅读
目的观察增殖细胞核抗原(PCNA)小干扰RNA(siRNA)对人结肠癌CaCO_2细胞抑制作用。方法WST-8法和克隆形成抑制研究No.2和No.4 PCNA siRNA对CaCO_2细胞的作用;碘化丙锭(PI)单染检测细胞周期,Hochest33258染色观察凋亡形态,膜联蛋白(Annexin)V和PI双染测定凋亡百分率。结果No.2和No.4 PCNA siRNA与CaCO_2细胞作用,增殖抑制率分别为(72.24±1.01)%和(74.67±3.35)%;克隆形成抑制率均达90%;两种药物转染细胞均出现明显亚二倍体峰,Sub-G_1+G_0/G_1期减少,S+G_2/M期增多;siRNA处理组细胞有较多凋亡形态变化;凋亡率随浓度增加而增加,且早期凋亡率持续高于晚期凋亡/坏死率。结论PCNA siRNA显著抑制人结肠癌CaCO_2细胞增殖及克隆形成;细胞停留于G_2/M期,明显诱导细胞凋亡。
Objective To observe the inhibitory effect of proliferating cell nuclear antigen (PCNA) small interfering RNA (siRNA) on human colon carcinoma CaCO 2 cells. Methods The effect of No.2 and No.4 PCNA siRNA on CaCO_2 cells was studied by WST-8 and clonogenic assay. The cell cycle was detected by single stained with propidium iodide (PI) and the morphology of apoptotic cells was observed by Hochest33258 staining. Annexin ) V and PI double staining to determine the percentage of apoptosis. Results No.2 and No.4 PCNA siRNAs could inhibit the proliferation of CaCO 2 cells by 72.24 ± 1.01% and 74.67 ± 3.35%, respectively. The inhibitory rates of clonogenicity reached 90%. Both of the transfected cells Sub-G_1 + G_0 / G_1 phase decreased and S + G_2 / M phase increased. SiRNA-treated cells showed more apoptotic morphological changes; apoptosis rate increased with increasing concentration, Mortality continued to be higher than late apoptosis / necrosis. Conclusions PCNA siRNA can significantly inhibit the proliferation and colony formation of human colon cancer CaCO 2 cells. The cells remain in G 2 / M phase and induce apoptosis obviously.