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。
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.
出处
《陕西医学杂志》
CAS
2015年第2期134-137,共4页
Shaanxi Medical Journal
基金
陕西省科技攻关项目(2011k13-01-03)
