Long Non-coding RNA MEG3 Induces Renal Cell Carcinoma Cells Apoptosis by Activating the Mitochondria

来源 :Journal of Huazhong University of Science and Technology(med | 被引量 : 0次 | 上传用户:hflx152
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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-q PCR 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 293 T. 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 m RNA by RT-q PCR 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 m RNA 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. 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-q PCR 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 293 T. 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 m RNA by RT-q PCR in the transfected 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 m RNA level was declined by MEG3 overexpression (P <0.05). It was confirmed that MEG3 induces the apoptosis of RCC cells possibly by activating the mitochondrial pathway.
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