韩娜娜 徐细明△ 张美霞 甘园园 何晓琴.长链非编码RNA对HepG2 肝癌细胞增殖及凋亡的影响[J].现代生物医学进展英文版,2016,16(10):1829-1832. |
长链非编码RNA对HepG2 肝癌细胞增殖及凋亡的影响 |
Influence of Up-regulation of Long Non-coding RNA RP13-514E23.1 onProliferation and Apoptosis of Hepatocellular Carcinoma Cell Line |
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DOI: |
中文关键词: 肝癌 长链非编码RNA RP13-514E23.1 MAPK10 增殖 凋亡 |
英文关键词: Hepatocellular carcinoma Long non-coding RNA RP13-514E23.1 MAPK10 Proliferation Apoptosis |
基金项目:湖北省自然科学基金项目(2012FKC143) |
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中文摘要: |
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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