RNA干扰抑制骨肉瘤MG-63细胞β-catenin的表达

摘要

Objective To invcstigate the role of down-regulation of β-catenin by small interfering RNA(siRNA)on the survival,invasion and chemosensitivity of human MG-63 osteosarcoma cells. Methods The siRNA against β-catenin was constructed and transfected into MG-63 cells with LipofectamineTM 2000. The expression of β-catenin was dctected by RT-PCR and Western blot. Flow cytometry was used to detect the cell cycle. The proliferation of cells was assayed by CCK-8 method. Transwell test was used to detect the invasion ability of MG-63 cells. Chemosensitivity to doxorubicin of MG-63 cells was determined by CCK-8. Results As compared with the control group,the mRNA and protein level of β-catenin was significantly decreased in experimental group transfected with β-catenin siRNA. Cell cycle analysis revealed no obvious changes in phases in MG-63 cells. Cell invasion ability in experimental group was significantly decreased. CCK-8 assay revealed no change in proliferation of cells. Chemosensitivity to doxorubicin in experimental group was increased. Conclusion Knock-down of β-catenin gene may decrease the invasion ability and increase chemosensitivity to doxorubicin in MG-63 osteosarcoma cells.%目的 通过抑制Wnt信号通路核心蛋白β-catenin在骨肉瘤细胞系MG-63中的表达,探讨β-catenin对MG-63细胞的生长、细胞侵袭性以及耐药性等方面的作用.方法 设计合成β-catenin的siRNA序列,LipofectamineTM2000转染入MG-63细胞.用RT-PCR和Western blot法检测干扰后β-catenin的mRNA和蛋白表达情况,利用流式细胞仪检测细胞周期,Transwell法检测细胞侵袭能力,CCK-8法检测细胞增殖,利用阿霉素检测细胞耐药性.结果 干扰后,实验组细胞的β-catenin mRNA和蛋白表达较对照组和空白组显著降低(均P＜0.05);细胞周期检测实验组与对照组和空白组比较差异无统计学意义(P＞0.05);Transwell法结果显示实验组细胞侵袭能力较对照组和空白组显著下降(均P＜0.05);CCK-8法检测显示实验组细胞增殖力较对照组和空白组略有降低,但差异无统计学意义(均P＞0.05);多柔比星敏感性检测显示实验组较对照组和空白组细胞生长抑制率显著增加(均P＜0.05).结论 特异性抑制β-catenin基因表达可抑制骨肉瘤MG-63细胞的侵袭能力,并增加MG-63细胞对化疗药物阿霉素的敏感性.