RNA干扰抑制BMP-2基因表达对人肝癌SMMC7721细胞增殖和凋亡的影响

BMP-2-siRNA Mediated Apoptosis and Proliferation of Human Hepatoma Cell Line SMMC7721

目的设计并化学合成针对骨形态发生蛋白2(BMP-2)的siRNA分子片段,转染人肝癌SMMC7721细胞,观察其对SMMC7721细胞增殖和凋亡的影响。方法阳离子脂质体法瞬间转染SMMC7721细胞,半定量逆转录聚合酶链反应(RT-PCR)法和Western印迹法检测转染BMP-2-siRNA后细胞BMP-2mRNA水平和蛋白水平的变化,MTT法和流式细胞术检测转染BMP-2-siRNA后SMMC7721细胞增殖、细胞周期和凋亡的变化。结果 3对特异性BMP-2-siRNA均有效地抑制了BMP-2基因的表达,以siRNA-B抑制效果最好。转染BMP-2-siRNA后SMMC7721细胞的增殖能力明显受到抑制(P<0.05)且明显促进了SMMC7721细胞的凋亡。结论靶向BMP-2基因的siRNA分子片段可以有效地抑制人肝癌SMMC7721细胞的增殖并促进其凋亡。

Objective To design and synthesis siRNAs targeting Bone Morphogenetic Protein2（BMP-2）gene and to observe its influence on the proliferation and apoptosis of SMMC7721 cells.Methods Three specific siRNAs of BMP-2 were designed and synthesized,and were transiently transfected into SMMC7721 via cathodolyte liposome transfection method.Reverse transcriptase PCR and Western blotting were used to measure BMP-2 expression at mRNA and protein level,respectively.The cell proliferation was assessed by MTTassay and the changes in cell cycle and apoptosis were evaluated by flow cytometry.Results All the 3 specific BMP-2-siRNA（A,B,C）could effectively inhibit the expression of BMP-2 gene,BMP-2-siRNA-B had the highest inhibitive effect.The proliferation of SMMC7721 cells was markedly inhibited and the apoptosis of SMMC7721 cells was increased transfected by BMP-2-siRNAs（P0.05）.Conclusion The siRNA of BMP-2 could effectively inhibit the expression of BMP-2 and induce the growth inhibition and apoptosis increase in SMMC7721 cells.