【摘要】 目的:观察应用RNA干扰(RNAi)技术敲低PIK3R1表达后在体外对人类多发性骨髓瘤细胞系RPMI8226细胞侵袭的抑制作用。方法:将重组质粒表达载体pGenesil-1.1-PIK3R1转染至RPMI8226细胞。Realtime PCR和Western blot分别检测转染前后目的基因mRNA和蛋白的表达水平,酶联免疫吸附试验(ELISA)检测细胞外基质金属蛋白酶(MMP)-2、MMP-9的浓度变化。Transwell实验评价肿瘤细胞侵袭能力的变化。结果:pGenesil-1.1-PIK3R1可以有效抑制PIK3R1mRNA及蛋白的表达,MMP-2、MMP-9表达下调,基质金属蛋白酶组织抑制因子(TIMP)-2表达上调,ELISA证实细胞外MM9-2、MMP-9浓度在治疗组明显减低。Tramwell穿过细胞数:对照组(115.5±3.9)和无义序列组(112.8±6.0),pGenesil-1.1-PIK3R1转染组(73.7±4.0)。结论:靶向PIK3RI的RNAi技术可以序列特异性地抑制PI3KR1表达,在体外明显抑制RPMI8226细胞的侵袭能力。更多还原

【Abstract】 Objective:To observe the inhibitory effects on the invasion and metastasis of Multiple Myeloma RPMI8226 cells line by small hairpin RNA targeting PIK3R1 in vitro.Methods:The recombinant plasmid vector expression vector which contained PIK3 Rl shRNA was transfected into RPMI8226 cells.Real-time PCR and Western blot were used to detect the expression of P1K3 Rl.ELESA was used to measure the change in the MMP-2/MMP-9expression.The invasion ability of the tumor cells was examined by Transwell tests.Resuits:pGenesil-1.1-PIK3R1 mediated shRNA targeting PIK3R1 dramatically down-regulated their expression in RPMI8226 cells.MMP-2and MMP-9were downregulated,and TIMP-2was upregulated.The extracellular levels of MMP-2and MMP-9were decreased.Transwell showed that the number of cells invading through the matrigel in control,nonsense sequence.And pGenesil-1,1-PIK3R1 transfection groups were 115.5±3.9,112.8±6.0,and 73.7±4.0respectively.Conclusion:ShRNA targeting PIK3R1down-regulated significantly their expression in a sequence-specific manner,and inhibited the invasion of Multiple Myeloma RPMI8226 lcells in vitro.更多还原