沉默c-maf基因对多发性骨髓瘤细胞系RPMI8226增殖和侵袭力的影响

摘要

目的：探讨c-maf基因对多发性骨髓瘤细胞增殖、侵袭力的影响。方法：脂质体法用c-maf siRNA转染多发性骨髓瘤细胞系RPMI8226，RT-PCR技术检测c-maf基因的表达，MTT实验和Transwell小室分别检测多发性骨髓瘤细胞增殖活性和侵袭力的改变，流式细胞术检测细胞周期，Western blots检测相关蛋白的表达情况，并检测Caspase的活性。结果：c-maf siRNA有效地转染入细胞并抑制了c-maf基因的表达。转染c-maf siRNA细胞的增殖活性及体外侵袭力均显著下降（P<0.05），细胞周期阻滞在G2/M期，survivin、MMP-2、MMP-9、ARK5、cyclin B1蛋白水平明显低于对照组（P<0.05），Caspase-3/7蛋白活性高于对照组（P<%Objective:To investigate the effect of c-maf gene on the MM cells' proliferation and invasion activity. Methods:Lipo-fectin Reagent was used to transfect c-maf siRNA into multiple myeloma cell of RPMI8226. The mRNA expression level of c-maf was detected by RT-PCR.Cell growth curve was measured by MTT assay. Transwell chamber test was used to measure MM cells' in vitro in-vasion activity. The cell cycle distribution were assessed by flow cytomentry. The protein expression levels of survivin,MMP-2, MMP-9, ARK5 and cyclin B1 were detected by Western blot. We also detected the activity of Caspase-3/7. Results:The c-maf siRNA was effectively transfected into cells and the mRNA expression of the c-maf gene was inhibited.MTT test and Transwell chamber test showed that the proliferation and in vitro invasion activity of transfected cells were significantly lower than those of other two groups (P<0.05). Cell cycle of c-maf siRNA transfected group cells was arrested in G2/M phase. The expression levels of survivin,MMP-2, MMP-9,ARK5,cyclin B1 and the activity of Caspase-3/7 between c-maf siRNA transfected group and the other two groups were sta-tistically different (P<0.05). Conclusion:c-maf gene by c-maf siRNA can remarkably inhibit proliferation and invasion of multiple my-eloma cell lines of RPMI8226. C-maf gene may be used as the target for multiple myeloma gene therapy.