miR-203通过靶向FABP4抑制肺癌细胞的转移

摘要

目的 探讨脂肪酸转运蛋白4(FABP4)对肺癌细胞转移能力的影响并研究靶向FABP4的miRNA.方法 通过ELISA和western blot检测不同转移能力的肺癌细胞FABP4表达情况.通过shRNA减少FABP4表达或慢病毒过表达FABP4,观察其对肺癌细胞转移能力的影响.通过miRNA数据网站预测靶向FABP4的miRNA,并用Q-PCR检测其在不同转移能力的肺癌细胞的表达情况.结果 高转移能力的NL9980、H661、95C肺癌细胞中FABP4的表达较低转移能力的L9981、A549、PC13显著增加(P<0.05).降低NL9980细胞中FABP4的表达,其转移能力被显著抑制(P<0.05);而过表FABP4后,A549转移能力显著增强(P<0.05).miR-203、miR-361和miR-539在高转移能力的肺癌细胞中表达较低转移的显著减少(P<0.05),并且miR-203抑制NL9980细胞FABP4表达的作用最强.过表达靶向FABP4位点突变的FABP4蛋白,可以显著减少miR-203对NL9980细胞转移能力的抑制(P<0.05).结论 FABP4可以促进肺癌细胞的转移;而miR-203可靶向抑制FABP4表达,进而抑制肺癌细胞的转移.%Objective To explore the role of fatty acid binding protein 4 (FABP4) in regulating lung cancer cell metastasis and identify miRNAs that target FABP4.Methods The expression of FABP4 in lung cancer cells with different metastatic potentials was detected using enzyme-linked immunosorbent assay(ELISA)and Western blotting.The effects of FABP4 knockdown or overexpression by shRNA or a recombinant lentivirus, respectively, on lung cancer cells metastasis were assessed. The miRNAs that targeted FABP4 were screened using target prediction algorithms and the results were verified with Q-PCR. Results FABP4 expression was significantly higher in lung cancer cell lines with high metastatic potentials(NL9980,H661,and 95C) than in those with low metastatic potentials (L9981, A549, and PC13) (P<0.05). FABP4 knockdown in NL9980 cells resulted in significantly inhibited metastasis of the cells (P<0.05), while FABP4 overexpression obviously promoted the metastasis of A549 cells (P<0.05). The expressions of miR-203, miR-361 and miR-539 were significantly higher in highly metastatic lung cancer cells than in the cells with low metastatic potentials (P<0.05). In NL9980 cells, FABP4 expression was most obviously suppressed by miR-203(P<0.05),and target site mutational FABP4 overexpression significantly attenuated the inhibitory effect of miR-203 on NL9980 metastasis (P<0.05). Conclusion FABP4 can promote lung cancer metastasis, and by targeting FABP4 to inhibit its expression,miR-203 can suppress the metastasis of lung cancer cells.