shRNA沉默IGF-Ⅱ基因对人肾上腺皮质癌细胞株SW-13增殖与凋亡的影响

Effects of IGF-Ⅱ gene silencing by shRNA on proliferation and apoptosis of human adrenocortical tumor cell line SW-13

目的探讨shRNA沉默IGF-Ⅱ基因对人肾上腺皮质癌细胞株SW-13增殖与凋亡的影响。方法构建针对人IGF-Ⅱ基因的shRNA慢病毒表达载体,稳定转染人肾上腺皮质癌细胞株SW-13。将实验随机分为3组:空白对照组、阴性对照组、IGF-Ⅱ-shRNA组,应用RT-PCR检测转染后各组IGF-Ⅱ基因mRNA的表达,以MTT比色法、流式细胞术检测转染后SW-13细胞生长增殖、凋亡的情况。结果 IGF-Ⅱ-shRNA组的IGF-Ⅱ基因mRNA表达量明显低于阴性对照组和空白对照组(P均<0.01),IGF-Ⅱ-shRNA组较之其他两组能明显抑制细胞的增殖(P均<0.01),IGF-Ⅱ-shRNA组的细胞凋亡率与其他两组相比显著增加(P均<0.01)。结论靶向IGF-Ⅱ基因的shR-NA可以有效抑制人肾上腺皮质癌细胞株SW-13的生长,并促进其凋亡,IGF-Ⅱ基因可能是肾上腺皮质癌治疗的一个潜在靶点。

Objective To investigate the effect of IGF- Ⅱ gene silencing by shRNA on growth and apoptosis in human adrenocortical tumor cell line SW-13. Methods ShRNA lentiviral vector that can express shRNA of IGF- Ⅱ was constructed, and transfected into human adrenocortical tumor cell line SW-13. SW-13 cells were divided into three groups： blank control group, negative group and experimental group. After transfection, the mRNA was detected by Reverse transcriptionpolymerase chain reaction. Cell proliferation and apoptosis were determined with MTT and flow cytometry. Results Compared with blank control group and negative group, the expression of IGF- Ⅱ mRNA was decreased significandy in IGF- Ⅱ - shRNA group （ all P 〈 0.01 ） ; cell proliferation was significantly inhibited in IGF- Ⅱ -shRNA group（ all P 〈 0.01 ） ; cell apoptosis was significantly higher in IGF- Ⅱ -shRNA group than in blank control group and negative group （ all P 〈 0.01 ）. Conclusions The shRNA of IGF- Ⅱ gene can effectively inhibit SW-13 cell proliferation and promote cell apoptosis. IGF- Ⅱ might be a potential target for the therapy of adrenal cortical carcinoma.