摘要
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 explore the inhibitory effect of proliferating cell nuclear antigen （ PC- NA） small interfering RNA （siRNA） on human colon carcinoma CaCO2 cells. Methods WST-S 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 hochest33258 staining. AnnexinV-PI dying was applied to detect the percentage of apoptotic cells. Results After activation of either No. 2 or No. 4 PCNA siRNA with CaCO2 cells, 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 cells all displayed Sub-G1 apeptotic peaks obviously,reduction at Sub-G1 ＋ G0/G1 phase and the increase at S ＋ G2/M phase; More apoptotic morphologicgl 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 apoptotic cells was continuously higher thatt late apoptotic and necrosis cells. Conclusion PCNA siRNA significantly inhibited human colon CaCO2 cell proliferation and colony formation, which was contributed to the cell cycle arresting at G2/M phase, and predominant induction of cell apoptosis.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2007年第4期454-456,F0003,共4页
Chinese Journal of Experimental Surgery
基金
广州市科技计划资助项目(2002J1-C0361)
