过表达miR-200家族对α粒子诱发恶性转化人支气管上皮细胞迁移能力的影响

Effects of miR-200 family overexpression on migration of malignantly transformed human bronchial epithelial cells induced by α-particles

目的:探讨过表达miR-200家族对α粒子诱发恶性转化人支气管上皮细胞(RHT35细胞)迁移能力的影响。方法:用SiPORT转染试剂转染100n mol/LmiR-200b mimic和miR-141 mimic或阴性对照(mimic control)于RHT35细胞中72h后,分别采用免疫印迹、实时荧光定量PCR技术检测过表达miR-200b和miR-141对RHT35细胞ZEB家族(ZEB1和ZEB2)及上皮间质转变相关标志分子表达的影响;并用伤口愈合实验研究过表达miR-200b和miR-141对RHT35细胞迁移能力的影响。结果:ZEB1和ZEB2蛋白在RHT35细胞中高表达。与阴性对照转染组相比,miR-200b mimic和miR-141 mimic转染组中miR-200靶基因ZEB家族表达显著下调(P<0.01);而上皮间质转变相关标志分子E-cad mRNA表达显著上调(P<0.05或P<0.01),N-cad表达显著下调(P<0.01)。与阴性对照转染组相比,miR-200b mimic和miR-141mimic转染组能显著抑制RHT35细胞的迁移能力。结论:过表达miR-200b和miR-141对RHT35细胞的迁移能力有显著抑制作用。

OBJECTIVE： To explore effects of miR-200 family overexpression on migration of malignantly transformed human bronchial epithelial cells （RHT35） induced by α-particles. METHODS：The mRNA and protein levels of ZEB family （ZEB1 and ZEB2） and epithelial-mesenchymal transition marker in RHT35 was measured by Real Time PCR and western blot 72 h after transfection of 100 nmol/L miR-200b mimic, miR-141 mimic or mimic control into RHT35 cells with SiPORT reagent. In vitro wound healing assay was used to investigate whether overexpression of miR-200b and miR-141 could block migration of malignantly transformed cells induced by α-particles. RESULTS：The protein levels of ZEB1 and ZEB2 in RHT35 cells were upregulated compared with R15H20 and BEP2D cells. Overexpression of miR-200b and miR-141 led to reduced expression of ZEB1 and ZEB2 in RHT35 cells compared with mimic control （P0.01）. Overexpressions of miR-200b and miR-141 upregulated E-cad expression （P0.05 or P0.01） and downregulated N-cad expression in RHT35 cells （P0.01）. Overexpressions of miR-200b and miR-141 markedly blocked migration of RHT35 cells compared with mimic control. CONCLUSION：Overexpressions of miR-200b and miR-141 strongly block migration of RHT35 cells.