miRNA-149抑制膀胱癌细胞侵袭的初步分子机制研究

Study on the preliminary molecular mechanism of miRNA-149 inhibiting bladder cancer cell invasion

目的探讨miRNA-149抑制膀胱癌细胞侵袭的可能分子机制。方法采用PolyJet^( TM)DNA In Vitro Tranfection Reagent将miRNA-149过表达质粒稳定转染膀胱癌细胞T24、T24T,并用荧光定量PCR技术检测转染率;利用免疫蛋白印迹法检测EMT标志蛋白及miRNA-149下游靶基因IGFBP5和PDGFRA蛋白的表达。结果在T24、T24T细胞中成功过表达miRNA-149,且差异具有统计学意义(P<0.05)。miRNA-149显著抑制膀胱癌细胞EMT发生,从而抑制膀胱癌细胞侵袭。进一步机制研究发现miRNA-149可抑制其下游靶基因IGFBP5和PDGFRA蛋白的表达。结论该研究表明miRNA-149抑制膀胱癌侵袭可能与EMT的发生及其下游基因IGFBP5和PDGFRA的表达密切相关,为临床膀胱癌的转移研究提供了新的启示。

Objective To investigate the possible molecular mechanism of miRNA-149 inhibiting the invasion of bladder cancer cells. Methods miRNA-149 overexpression plasmid was stably transfected into bladder cancer cells T24, T24T using PolyJetTM DNA In Vitro Tranfection Reagent, and the transfection rate was detected by real-time PCR; the expression of EMT marker protein and miRNA-149 downstream target genes IGFBP5 and PDGFRA proteins were detected by Western blot method. Results miRNA-149 was successfully overexpressed in T24 and T24T cells, and the difference was statistically significant（P〈0.05）. miRNA-149 significantly inhibited the occurrence of EMT in bladder cancer cells, thereby inhibiting the invasion of bladder cancer cells. Further mechanism studies found that miRNA-149 inhibited the expression of its downstream target genes IGFBP5 and PDGFRA proteins. Conclusion This study demonstrates that miRNA-149 inhibition of bladder cancer invasion may be closely related to the occurrence of EMT and the expression of downstream genes IGFBP5 and PDGFRA, providing new insights for the study on clinical bladder cancer metastasis.