摘要
目的检测雷帕霉素刺激SCC-090后miR-181c-5p的表达量,验证后者与自噬基因Beclin1的作用关系及对SCC-090的增殖与凋亡的影响。方法应用qRT-PCR检测雷帕霉素(RAPA)及3-甲基腺嘌呤(3-MA)分别刺激SCC-090细胞后miR-181c-5p的表达量;应用qRT-PCR、免疫印迹及荧光活性验证miR-181c-5p与Beclin1的作用关系;应用CCK8检测细胞的增殖,应用流式细胞术检测凋亡细胞比率,应用Western blot检测凋亡基因Bcl-2的表达。结果雷帕霉素处理细胞后,miR-181c-5p表达升高(P<0.05),3-MA处理后的细胞中,miR-181c-5p的表达下降(P<0.05);当miR-181c-5p高表达时,Beclin1表达下调(P<0.05),当miR-181c-5p表达降低时,Beclin1表达升高(P<0.05);miR-181c-5p与Beclin1有靶向关系;miR-181c-5p可明显抑制SCC-090细胞的增殖并促进细胞的凋亡。结论雷帕霉素诱导SCC-090细胞自噬后miR-181c-5p的表达量明显升高,且miR-181c-5p可通过抑制Beclin1的表达降低SCC-090细胞的增值能力且促进细胞的凋亡。
Objective To investigate the expression of miR-181c-5p of in SCC-090 after rapamycin(RAPA)stimulus,and to verify the correlation between the latter and the autophagy gene Beclin1.Methods The expression levels of miR-181c-5p in SCC-090 cells stimulated by RAPA and 3-methyl adenine(3-MA)respectively were detected by qRT-PCR.Western Blot and luciferase activity assay were used to verify the correlation between miR-181c-5p and Beclin1.The proliferation of SCC-090 cells was detected by CCK8,and the apoptotic rate was detected by Flow cytometry,and the expression levels of Bcl-2 were detected by Western Blot.Results After RAPA treatment,the expression levels of miR-181c-5p were significantly increased,however,after 3-MA treatment,which were were significantly decreased(P<0.05).When the expression levels of miR-181c-5p were increased,the expression levels of Beclin1 were decreased,when the expression levels of miR-181c-5p were decreased,the expression levels of Beclin1 were increased.There was a target correlation between miR-181c-5p and Beclin1,and miR-181c-5p could obviously inhibit the growth and promote the apoptosis of SCC-090 cells.Conclusion The expression levels of miR-181c-5p are increased significantly after autophagy of SCC-090 cells induced by RAPA,and miR-181c-5p can reduce cell proliferation and promote apoptosis of SCC-090 cells by inhibiting the expression of Beclin1.
作者
刘军刚
刘洋
哈庆
LIU Jungang;LIU Yang;HA Qing(Department of Stomatology,Beijing Rehabilitation Hospital Affiliated to Capital Medical University,Beijing 100144,China;不详)
出处
《河北医药》
CAS
2021年第18期2734-2737,共4页
Hebei Medical Journal