Long non-coding RNA MEG3 induces renal cell carcinoma cells apoptosis by activating the mitochondrial pathway
Long non-coding RNA MEG3 induces renal cell carcinoma cells apoptosis by activating the mitochondrial pathway
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DOI：
10.1007/s11596-015-1467-5
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发表时间：
2015
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通讯作者：
Miao Wang 王淼;Tao Huang 黄韬;Gang Luo 罗刚;Chao Huang 黄超;Xing-yuan Xiao 肖行远;Liang Wang 汪良;Guo-song Jiang 蒋国松-Guo-song-Jia
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作者：
Miao Wang 王淼;Tao Huang 黄韬;Gang Luo 罗刚;Chao Huang 黄超;Xing-yuan Xiao 肖行远;Liang Wang 汪良;Guo-song Jiang 蒋国松-Guo-song-Jia
本研究旨在探讨长链非编码RNA（LncRNA）MEG3对肾细胞癌（RCC）细胞786-0生物学行为的影响及其可能的机制。通过 RT-qPCR 检测 29 名 RCC 患者的肿瘤组织和邻近非肿瘤组织以及 RCC 系 786-0 和 SN12 以及人胚肾细胞系 293T 中的 MEG3 表达水平。使用lipofectamine 2000将质粒GV144-MEG3（MEG3过表达质粒）和GV144（对照质粒）稳定转染至786-0细胞中。MTT法测定转染细胞中的细胞活力，PE膜联蛋白V和7AAD染色后流式细胞仪测定细胞凋亡率，Western blotting测定凋亡相关蛋白表达，RT-qPCR测定转染细胞中Bcl-2 mRNA。结果显示，MEG3在RCC组织（P<0.05）和RCC细胞系（P<0.05）中明显下调。 GV144-MEG3转染24 h以上后786-0细胞活力明显下降（P<0.05）。一致地，转染GV144-MEG3 48 h的786-0细胞凋亡率显着增加（P<0.05）。此外，过表达MEG3可降低Bcl-2和procaspase-9蛋白的表达，增强Cleaved caspase-9蛋白的表达，促进细胞色素c蛋白向细胞质的释放（P<0.05）。此外，MEG3过表达使Bcl-2 mRNA水平下降（P<0.05）。结论MEG3可能通过激活线粒体途径诱导RCC细胞凋亡。
This study aimed to examine the effect of long non-coding RNA (LncRNA) MEG3 on the biological behaviors of renal cell carcinoma (RCC) cells 786-0 and the possible mechanism. MEG3 expression levels were detected by RT-qPCR in tumor tissues and adjacent non-tumor tissues from 29 RCC patients and in RCC lines 786-0 and SN12 and human embryonic kidney cell line 293T. Plasmids GV144-MEG3 (MEG3 overexpression plasmid) and GV144 (control plasmid) were stably transfected into 786-0 cells by using lipofectamine 2000. Cell viabilities were determined by MTT, cell apoptosis rates by flow cytometry following PE Annexin V and 7AAD staining, apoptosis-related protein expressions by Western blotting, and Bcl-2 mRNA by RT-qPCR in the transfected cells. The results showed that MEG3 was evidently downregulated in RCC tissues (P<0.05) and RCC cell lines (P<0.05). The viabilities of 786-0 cells were decreased significantly after transfection with GV144-MEG3 for over 24 h (P<0.05). Consistently, the apoptosis rate was significantly increased in 786-0 cells transfected with GV144-MEG3 for 48 h (P<0.05). Furthermore, overexpression of MEG3 could reduce the expression of Bcl-2 and procaspase-9 proteins, enhance the expression of cleaved caspase-9 protein, and promote the release of cytochrome c protein to cytoplasm (P<0.05). Additionally, Bcl-2 mRNA level was declined by MEG3 overexpression (P<0.05). It was concluded that MEG3 induces the apoptosis of RCC cells possibly by activating the mitochondrial pathway.