Abstract:

Objective To explore the expression of miRNA-135a（miR-135a）in the hepatic carcinoma and its clinical significance. Methods The level of expression of miR-135a in the hepatic carcinoma tissues of 20 patients with hepatic carcinoma and the commensurate adjacent normal liver tissues were detected by real?time quantitation PCR. HepG2 cells were transfected with miR?135a mimics mediated by liposomal transfection reagent and the expression of miR-135a in the transfected HepG2 cells were detected by real?time quantitation PCR. The changes of cell clone formation rat and cell viability were analyzed by tablet clone formation experiment（PCF）and MTT method, respectively. Meanwhile, the expression of the related genes of miR?135a regulation were analyzed to discover the role of miR?135a in maintain normal function. Results The miR-135a was detected in both hepatic carcinoma and commensurate adjacent normal liver tissues. The expression of miR-135a in the commensurate adjacent normal liver tissues were significantly higher than that in the hepatic carcinoma （P<0.05）. Compared with negative control, the expression of miR-135a in the miR?135a mimics transfected HepG2 cells was significantly increased （P<0.05）, however, the clone formation rate and viability were significantly decreased （P<0.05）. And the expression of miR-135a regulation related genes including FOS, PI3, Jak2, Stat3 were all down regulated（P<0.05）. These data suggested that the up?regulated of miR-135a could inhibited the clone formation rate and decrease the cell viability. Conclusion miR-135a was differently expression in hepatic carcinoma and normal liver tissues and the up-regulated of miR-135a could inhibited the clone formation rate and decrease the cell viability. miR-135a contributed to the viability and occurrence of hepatic carcinoma by regulated of the related genes directly or indirectly, miR?135a may became the important target for clinical treatment and prognosis of hepatic carcinoma.

Key words: miR?135a, HepG2, Hepatic carcinoma, miR?135a mimics

摘要：

 目的 探讨miRNA?135a（miR?135a）在肝癌组织中的表达及其临床意义。方法 采用实时荧光定量PCR检测20例肝癌组织及其癌旁组织中miR?135a的表达水平, 并通过脂质体介导的方法将miR?135a模拟物（miR?135amimics）转染肝癌细胞株HepG2，采用实时荧光定量PCR测定转染后细胞中miR?135a的表达水平；通过平板克隆实验和MTT法分别检测转染后细胞克隆形成率和存活率的变化，并对miR?135a调控相关靶细胞的基因表达水平进行检测，观察miR?135a在肝癌细胞维持正常功能中的作用。结果 通过对20例肝癌组织及其癌旁组织中miR?135a的表达水平进行检测，结果miRNA?135a的表达在正常组织中相对更高，且两组之间存在显著性差异（P<0.05）；与阴性对照组比较，转染miR?135amimics后HepG2细胞中miR?135a表达水平显著提高（P<0.05），而形成单克隆能力和细胞存活率均显著下降（P<0.05），同时转染组细胞中与miR?135a细胞调控相关靶基因FOS、PI3、Jak2、Stat3的mRNA表达量相较于NC组均显著降低（P<0.05），表明miR?135a表达提高后抑制细胞形成单克隆能力并降低存活率。结论 miR?135a在正常组织和肝癌组织中差异表达，同时当miR?135a表达提高时抑制肝癌细胞形成单克隆能力和降低细胞存活率，表明miR?135a可能通过一些相关靶基因直接或间接调控HepG2肝癌细胞的存活，在肝癌的发生过程中可能起到作用，miR?135a可能成为临床治疗肝癌和预后的重要靶点。

关键词: HepG2 细胞, 肝癌, miR?135a, miR?135a mimics