目的观察Prdx2基因对结直肠癌细胞SW480增殖及凋亡的影响并初步探讨其可能的机制。方法将干扰片段(Prdx2.shRNA)及空载体(阴性对照)经慢病毒转染结直肠癌细胞株SW480,1.25μg/ml嘌呤霉素筛选,建立稳定细胞株,以野生型SW480细胞作为空白对照。采用MTT法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,qRT-PCR及Westernblotting检测Prdx2及survivin的mRNA和蛋白表达情况。结果沉默Prdx2基因对结直肠癌细胞株SW480的生长和增殖有明显抑制作用。与阴性对照(s.59%±o.63%)及空白对照(5.79%±o.60%)比较,Prdx2基因沉默后,细胞凋亡率明显增加(26.60%±4.39%,P〈0.05)。qRT-PCR、Westernblotting结果显示,干扰Prdx2基因明显降低了survivinmRNA及蛋白表达。结论沉默Prdx2基因可抑制结直肠癌细胞增殖,增加凋亡,其机制可能与下调survivin的表达有关。
Objective To study the effect of Prdx2 gene on the proliferation and apoptosis of colorectal cancer cell line SW480 and its mechanism. Methods Colorectal cancer cell line SW480 was transfected with Prdx2-shRNA or empty vector respectively by means of lentivirus, and puromycin (1.25μg/ml) was used for screening. A stable cell line was established. Cells transfected with empty vector and wild-type cells were used as controls. Cell growth curve were determined by MTT method, and cell apoptosis was determined by flow cytometry. Both qRT-PCR and Western blotting were also used to detect the mRNA and protein expression of Prdx2 and survivin. Results Proliferation and apoptosis of colorectal cancer cell line SW480 were significantly inhibited by Prdx2 silent gene. Compared with control groups (5.59% ± 0.63%, 5.79% ± 0.60%), the apoptosis rate was significantly increased after Prdx2 gene silence (26.60% ±4.39%, P〈0.05). Western blotting and qRT-PCR analysis showed that survivin expression level was reduced due to Prdx2 gene interference. Conclusion Prdx2 gene silencing could inhibit the proliferation and increase the apoptosis of colorectal cancer cells, and it may be related to the down-regulation of survivin expression.