微小RNA在肝癌耐药Bel-7402/5-FU细胞中的表达分析

Analysis of miRNA in the Hepatocellular Carcinoma Resistant Cells Bel-7402/5-FU

目的:通过对比肝癌亲本Bel-7402细胞及其5-FU耐药Bel-7402/5-FU细胞中微小RNA(microRNA, miRNA)的表达差异,探讨miRNA在肝癌细胞多药耐药中的作用。方法:体外培养Bel-7402细胞及其耐药株Bel-7402/5-FU细胞,MTT法检测Bel-7402/5-FU细胞对不同化疗药物的敏感性,miRNA表达芯片检测Bel-7402细胞和Bel-7402/5-FU细胞miRNA的差异表达,qPCR验证差异表达的miRNA。结果:Bel-7402/5-FU细胞对不同的化疗药物均具有耐药性;miRNA芯片检测结果发现,耐药株Bel-7402/5-FU细胞与亲本Bel-7402细胞中,差异表达的miRNA有29个,其中上调14个,下调15个;qPCR验证结果发现,和亲本Bel-7402细胞比较,耐药株Bel-7402/5-FU细胞中miR-10a-5p、-371a-3p、-371a-5p、-372,-373-3p、-146-5p、-6723-5p显著下调,miR-4497、-3148显著上调。结论:肝癌多药耐药的发生与miRNA表达异常有关。

Objective: To determined the different experssion of miRNA in the parental cells Bel-7402 and its resistant cells Bel-7402/5-FU and investigated the effect of miRNA in the multidrug resistance of hepatocellular carcinoma. Methods: In vitro culture of Bel-7402 cells and its resistant strains Bel-7402/5-FU cells, MTT assays were used to detect the sensitivity to different chemotherapy drug of Bel-7402/5-FU. The miRNA array was used to detect the different expression between Bel-7402 cells and Bel-7402/5-FU cells. q PCR assay was used to verified the different miRNA. Results: Bel-7402/5-FU cells were resistant to diferent chemotherapy drug. In miRNA array analysis, 29 miRNAs were changed, and 14 miRNAs were up-regulated, 15 were down-regulated. The verified results of qPCR showed that compared with parental Bel-7402 cells, miR-10 a,-371 a-3 p,-371 a-5 p,-372,-373-3 p,-146-5 p,-6723-5 p were down-regulated in Bel-7402/5-FU cells, miR-4497 and-3148 were up-regulated in Bel-7402/5-FU cells. Conclusions: The diferent expression of miRNA was related to the MDR of HCC.