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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HepG2 增殖、凋亡的影响。方法：通过荧光定量PCR（qRT-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HepG2 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 pcDNA3.1-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HepG2细胞的增殖、凋亡的变化。结果：LncRNA RP13-514E23.1 在大部分肝癌细胞系（HepG2, 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pcDNA-RP13-514E23.1 后，qRT-PCR 检测HepG2 细胞的RP13-514E23.1 和MAPK10 mRNA 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HepG2 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：LncRNA 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HepG2 细胞增殖受到抑制、凋亡增加。

英文摘要:

Objective:To investigate the expression profile of long non-coding RNA RP13-514E23.1 in hepatocellular carcinoma cell lines, and to study the MAPK10 expression changes and the biological functions of the lncRNA in hepatocellular carcinoma cells HepG2, especially on cell proliferation and apoptosis.Methods:Quantitative reverse-transcription PCR(qRT-PCR) was used to detect the relative expression of RP13-514E23.1 in cell lines above. Transfect HepG2 cells with pcDNA-RP13-514E23.1 to up-regulate RP13-514E23.1 expression.The Mock group and NC group as control, to evaluated the efficiency of transfecting by qRT-PCR, and the expression of MAPK10 was evaluated by qRT-PCR and western blot. CCK-8 assay and flow cytometry were performed to evaluate the overexpression of RP13-514E23.1 on HepG2 cells' proliferation and apoptosis.Results:Compared with normal liver cell line LO2, there was a relatively low expression of RP13-514E23.1 in almost hepatocellular carcinoma cell lines (HepG2, SMMC, Huh7, Hep3B)(1.00 vs 0.58 ± 0.05, P<0.05). The expression of RP13-514E23.1 and MAPK10 were up-regulated in HepG2 cells after transfected pcDNA-RP13-514E23.1 (29.90 ± 1.40 and 2.42 ± 0.25 respectively, P<0.05), and the protein expression of MAPK10 was also up-regulated,(2.10± 0.16, P<0.05); proliferation of HepG2 cells was induced but apoptosis was enhanced ((5.53± 1.17)%, (6.40± 2.84)% and (46.87± 3.45)% respectively, P<0.01).Conclusion:The expression of RP13-514E23.1 is significantly down-regulated in almost hepatocellular carcinoma cell lines. After transfect HepG2 cells with pcDNA-RP13-514E23.1 to up-regulate RP13-514E23.1 expression, the expression of MAPK10 was up-regulated, proliferation of HepG2 cells were inhibited and apoptosis were processed.

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