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目的:探讨长链非编码RNA RP13-514E23.1在肝癌细胞系中的表达,并观察上调该基因后其下游基因MAPK10的变化及对肝癌细胞Hep G2增殖、凋亡的影响。方法:通过荧光定量PCR(q RT-PCR)检测RP13-514E23.1在正常肝细胞与肝癌细胞系中的表达差异,进一步构建质粒转染肝癌Hep G2细胞上调RP13-514E23.1的表达,以Mock组(只加转染试剂)和NC组(转染空载体pc DNA3.1-NC)作为对照评估转染效率及对MAPK10表达的影响。用CCK-8实验和流式细胞术检测转染前后Hep G2细胞的增殖、凋亡的变化。结果:Lnc RNA RP13-514E23.1在大部分肝癌细胞系(Hep G2,SMMC,Huh7,Hep3B)中的表达明显低于正常肝细胞(0.58±0.05 vs 1.00,P<0.05);转染pc DNA-RP13-514E23.1后,q RT-PCR检测Hep G2细胞的RP13-514E23.1和MAPK10m RNA表达量显著升高(分别为29.90±1.40、2.42±0.25,P<0.05),western blot检测MAPK10蛋白表达量较对照组也升高(2.10±0.16,P<0.05);CCK-8结果显示Hep G2细胞在各个时间段增殖均受到抑制(P<0.01),Mock组、NC组和实验组的凋亡率分别为(5.53±1.17)%、(6.40±2.84)%和(46.87±3.45)%(P<0.01)。结论:Lnc RNA RP13-514E23.1在肝癌细胞系中表达异常降低,上调其表达后MAPK10的表达升高,且Hep G2细胞增殖受到抑制、凋亡增加。
OBJECTIVE: To investigate the expression of RP13-514E23.1, a novel long-chain non-coding RNA, in human hepatocellular carcinoma cell line HCC and to investigate the effect of downregulation of MAPK10 on the proliferation and apoptosis of Hep G2 cell. Methods: The expression of RP13-514E23.1 in normal hepatocytes and hepatoma cell lines was detected by qRT-PCR. The expression of RP13-514E23.1 was further up-regulated in Hep G2 cells transfected with Mock group (transfection reagent only) and NC group (transfected empty vector pcDNA3.1-NC) as a control to evaluate transfection efficiency and the impact of MAPK10 expression. The proliferation and apoptosis of Hep G2 cells before and after transfection were detected by CCK-8 assay and flow cytometry. Results: The expression of Lnc RNA RP13-514E23.1 in most hepatocellular carcinoma cell lines (Hep G2, SMMC, Huh7, Hep3B) was significantly lower than that in normal hepatocytes (0.58 ± 0.05 vs 1.00, P <0.05) -RP13-514E23.1, the expression of RP13-514E23.1 and MAPK10m RNA in Hep G2 cells was significantly increased by q RT-PCR (29.90 ± 1.40,2.42 ± 0.25, P <0.05, respectively), and MAPK10 The expression of CCK-8 in Hep G2 cells was higher than that in the control group (2.10 ± 0.16, P <0.05). The results of CCK-8 showed that the proliferation of Hep G2 cells was inhibited at all time points (P <0.01) The apoptotic rates were (5.53 ± 1.17)%, (6.40 ± 2.84)% and (46.87 ± 3.45)%, respectively (P <0.01). CONCLUSION: The abnormal expression of Lnc RNA RP13-514E23.1 in hepatocellular carcinoma cell line was down-regulated. The up-regulated expression of MAPK10 increased Hep G2 cell proliferation and increased apoptosis.