【摘要】 目的观察增殖细胞核抗原（PCNA）小干扰RNA（siRNA）对人结肠癌CaCO₂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CaCO₂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CaCO₂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-G₁+G₀/G₁期减少,S+G₂/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CaCO₂细胞增殖及克隆形成;细胞停留于G₂/M期,明显诱导细胞凋亡。更多还原

【Abstract】 Objective To explore the inhibitory effect of proliferating cell nuclear antigen（PC- NA）small interfering RNA（siRNA）on human colon carcinoma CaCO₂ ceils.Methods WST-8 assay and colony formation inhibitory assay were employed to detect the inhibitory effect of No.2 and No.4 PC- NA siRNA on CaCO₂ cells.Flow cytometry was used to examine the cell cycle using propidium iodide（PI） dying.Morphological changes of apoptotic cells were observed under a fluorescent microscope with ho- chest33258 staining.AnnexinV-PI dying was applied to detect the percentage of apoptotic ceils.Results After activation of either No.2 or No.4 PCNA siRNA with CaCO₂ ceils,the proliferating inhibitory rates were（72.24±1.01）% and（74.67±3.35）%,respectively and the inhibitory rates of colony formation all reached to 90%;After transfected with the two agents respectively,the CaCO2 ceils all displayed Sub- G₁ apoptotic peaks obviously,reduction at Sub-G₁+G₀/G₁ phase and the increase at S + G₂/M phase; More apoptotie morphological changes were observed in both siRNA treatment groups.The apoptotic rate was increased in consistent with the increasing concentration of No.2 siRNA.The percentage of early apop- totic cells was continuously higher than late apoptotic and necrosis ceils.Conclusion PCNA siRNA sig- nificantly inhibited human colon CaCO₂ cell proliferation and colony formation,which was contributed to the ceil cycle arresting at G₂/M phase,and predominant induction of cell apoptosis.更多还原