摘要
目的探讨HER2基因高效RNA干扰质粒转染到SK-BR-3乳腺癌细胞后,对细胞的增殖及其凋亡的影响。方法采用脂质体转染法将针对HER2基因的高效RNA干扰质粒转染至乳腺癌细胞SK-BR-3中,通过细胞计数、MTT比色法、流式细胞术和Western blot检测PCNA增殖细胞核抗原,分析检测其对SK-BR-3细胞增殖、凋亡的影响。结果 MTT比色测定显示,HER2-shRNA2干扰质粒转染到SK-BR-3细胞后在48 h后现抑制,与空白对照组比抑制率分别为48 h 16.53%7、2h 39.03%9、6h 65.47%。流式细胞分析HER2-shRNA组细胞在96 h凋亡率达17.36%,明显高于阴性对照组的1.41%和空白对照组的1.25%(P<0.05)。结论将RNA干扰质粒HER2-shRNA2转染至SK-BR-3细胞中,可特异性地抑制SK-BR-3细胞的增殖并诱导其凋亡,为乳腺癌的靶向治疗奠定了基础。
Objective To examine the biologic effects of HER2 gene silencing on SKBR3 cells through the transfection of HER2-specific shRNA expression vector.Methods SK-BR-3 breast cancer cells overexpressing the HER2 gene were transfected with HER2-shRNA2 vector using Lipofectamine2000.After transfection the antiproliferative effect of HER2-shRNA2 was detected by MTT assay and apoptosis percentage was detected by flow cytometry.The expressions of proliferating cell nuclear antigen（PCNA） were evaluated by Western blot analysis.Results After transfection with HER2-shRNA2,inhibitory ratio of HER2-shRNA2 group was 16.53%（48h）,39.03%（72 h）,or 65.47%（96 h）.After Seventy-two hours,apoptosis percentage of SK-BR-3 cells of transfected with HER2-shRNA2 was increased to 10.29% compared to that in negative control and blank control groups.Conclusion Silencing of HER2 gene by shRNA can inhibit proliferation and induce apoptosis in SK-BR-3 cells.These promising results suggest that shRNA targeted against human HER2 may be valuable tools as antiproferative agents against neoplastic cells.
出处
《中国实验诊断学》
北大核心
2011年第5期802-805,共4页
Chinese Journal of Laboratory Diagnosis
基金
嘉兴市科技计划项目(2009AY2059)
