lncRNA IRAIN对肝癌细胞增殖和凋亡的影响及其机制-现代医学

lncRNA IRAIN对肝癌细胞增殖和凋亡的影响及其机制

单位：武汉市第一医院肝胆外科, 湖北武汉 430000

分类号：R735.7

出版年·卷·期（页码）：2020·39·第八期（943-948）

摘要：

目的：探讨长链非编码RNA（lncRNA）IRAIN对肝癌细胞增殖和凋亡的影响与机制。方法：采用RT-PCR检测IRAIN在42例肝癌组织和3种肝癌Hep 3B2.1-7、Huh7和HLE细胞中表达水平。将体外培养的Huh7细胞分为未感染组（正常培养）、Lv-Ctrl组（以携带空载对照的慢病毒感染）和Lv-IRAIN组（以携带IRAIN序列的慢病毒感染），采用RT-PCR检测各组细胞中IRAIN的表达水平，CCK-8法检测细胞增殖，流式细胞术检测细胞凋亡，Western blot检测细胞中Bax和Cleaved caspase-3蛋白的表达水平。结果：与癌旁组织相比，肝癌组织中IRAIN表达水平明显降低（P<0.05）；与人肝上皮THLE-3细胞相比，肝癌Hep 3B2.1-7、Huh7和HLE细胞中IRAIN表达水平均明显降低（P<0.05），且IRAIN在Huh7细胞中的表达水平最低。与未感染组比较，Lv-IRAIN组细胞增殖活力明显减弱，且细胞凋亡率和细胞中Bax、Cleaved caspase-3蛋白的表达水平均明显升高（P<0.05）；但Lv-Ctrl组与未感染组比较，各指标差异无统计学意义（P>0.05）。结论：IRAIN在肝癌中低表达，上调其表达可抑制细胞增殖并诱导细胞凋亡，其作用机制可能与上调Bax和Cleaved caspase-3蛋白的表达有关。

Objective: To investigate the effects and mechanisms of long-chain non-coding RNA (lncRNA) IRAIN on proliferation and apoptosis of hepatocellular carcinoma cells.Methods: The expression of IRAIN in 42 cases of hepatocellular carcinoma and 3 kinds of hepatocellular carcinoma Hep 3B 2.1-7, Huh 7 and HLE cells were detected by RT-PCR.Huh7 cells cultured in vitro were divided into uninfected group (normal culture), Lv-Ctrl group (infection with lentivirus carrying no-load control) and Lv-IRAIN group (infection with lentivirus carrying IRAIN sequence). The expression of IRAIN was detected by RT-PCR, cell proliferation was detected by CCK-8 method, cell apoptosis was detected by flow cytometry, expression level of Bax and Cleaved caspase-3 proteins were detected by Western blot.Results: Compared with the adjacent tissues, the expression of IRAIN in hepatocellular carcinoma was significantly lower (P<0.05); compared with human hepatic epithelial THLE-3 cells, the expression of IRAIN in Hep 3B 2.1-7, Huh 7 and HLE cells was significantly lower (P<0.05), and the expression of IRAIN in Huh 7 cells was the lowest. Compared with the uninfected group, the proliferation of Lv-IRAIN group was significantly decreased, and the apoptotic rate and the expression levels of Bax and Cleaved caspase-3 proteins in Lv-IRAIN group were significantly increased (P<0.05), but there was no significant difference in each index between Lv-Ctrl group and uninfected group (P>0.05).Conclusion: IRAIN is lowly expressed in hepatocellular carcinoma. Up-regulation of IRAIN expression can inhibit the proliferation of cancer cells and induce apoptosis, whose mechanism may be related to up-regulation of Bax and Cleaved caspase-3 protein expression.

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