摘要
目的以RNA干扰抑制血管平滑肌细胞(vascu lar smooth musc le cells,VSMCs)ORC1基因,探讨ORC1基因表达抑制后VSMCs增殖的变化。方法实验设置正常对照组、阴性siRNA组及阳性(ORC1+A、ORC1+B、ORC1+C)siR-NA组。应用W estern b lot检测ORC1基因表达的变化;应用MTT比色试验、3H-TdR掺入试验检测VSMCs增殖的情况。免疫细胞化学染色观察增殖细胞核抗原(proliferating cell nuc lear antigen,PCNA)表达。结果①siRNA转染后,3个阳性siRNA转染组ORC1基因表达水平均降低,尤以第2对阳性siRNA抑制效果最为显著,而空白对照组及阴性对照组间ORC1基因表达水平无显著差异。②siRNA转染使ORC1表达减弱后,VSMCs的MTT吸光度值3、H-TdR掺入量和PCNA表达量均较空白对照组及阴性对照组显著降低。结论RNA干扰介导的ORC1基因沉寂可显著抑制VSMCs增殖。
Objective To investigate the influence of RNA interference targeting ORC1 gene on the proliferation of rat vascular smooth muscle cells （VSMCss）. Methods VSMCss were transfected with siRNA targeting ORC1 gene by liposome. The expression of ORC1 protein was detected by Western blotting. MTT test and s H thymidine （ s H-TdR） incorporation were used to detect VSMCss proliferation. The expression of proliferating cell nuclear antigen （PCNA） in VSMCss was detected by immunocytochemistry. Results After transfected of the three pairs of siRNA targeting ORC1 genes respectively, the expression of ORC1 was all lower than that in the control group （non-transfection and negative siRNA）. Especially, that in the group transfected of the second pair positive siRNA decreased most significantly. The optical density of MTT, ^3H thymidine incorporation and the expression of PCNA decreased significantly in VSMCss transfected with siRNA targeting ORC1 gene as compared with that in the control group. Conclusion ORC1 gene silenced by RNA interference can inhibit VSMCss proliferation.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2006年第11期1161-1163,共3页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目(30470727)~~
