Let-7a-3p通过靶向AMPK对肺癌细胞系A549能量代谢及凋亡的影响  

The effects of let-7a-3p on energy metabolism and apoptosis of lung cancer cell line A549 by targeting AMPK

作　　者：张晶[1] 舒琦瑾 项宗汉 Zhang Jing

机构地区：[1]浙江省杭州市萧山区中医院,311200

出　　处：《浙江临床医学》2021年第5期617-620,共4页Zhejiang Clinical Medical Journal

基　　金：浙江省中医药管理局重点项目(2009ZA003)。

摘　　要：目的探讨转染Let-7a-3p对肺癌细胞能量代谢及凋亡影响及机制。方法根据肺癌A549细胞处理不同分为A549组、Let-7a-3p组(转染Let-7a-3p mimics)、阴性对照组(negative control mimic,NC)和未转染对照(non transfected,NT)组。采用RT-qPCR法检测各组细胞Let-7a-3p的水平,CCK8法检测细胞活力,海马细胞能量代谢实时测定仪XF分析测定氧耗率(OCR)和细胞外酸化率(ECAR),流失细胞术检测细胞凋亡,Western blot检测Let--7a-3p、Bcl-2、Caspase--3、Ras及AMPK蛋白的表达。结果Let-7a-3p组Let-7a-3p表达水平明显上调(P<0.01),而NC组Let-7a-3p表达水平与A549组比较,差异无统计学意义(P>0.05)。CCK-8法检测结果显示,与A549及NC组比较,Let-7a-3p组活细胞相对数明显减少(P<0.01)。能量代谢实时测定仪XF分析结果显示,与A549、NC组比较,转染Let-7a-3p后A549细胞的氧耗率(OCR)和细胞外酸化率(ECAR)减少(P<0.01)。流失细胞术检测结果显示,与A549组比较,Let-7a-3p组细胞凋亡率增多(P<0.01)。Western blot检测结果显示,过表达Let-7a-3p可以使A549细胞中Caspase-3与AMPK蛋白上调(P<0.01),而Bcl-2与Ras蛋白下调(P<0.01)。结论Let-7a-3p模拟物可以抑制肺癌细胞A549的活性和能量代谢并促进其凋亡。Let-7a-3p模拟物作用于癌细胞的机制可能是通过上调AMPK及Caspase-3蛋白水平,下调Bcl-2与Ras蛋白水平,从而触发能量代谢和凋亡通路,降低MMP,从而抑制ATP的产生。Objective The efect of Let-7a-3p on the energy metabolism and apoptosis of lung cancer cell line A549 is unknown.To investigate the effects and mechanisms of Let-7a-3p transfection on energy metabolism and apoptosis in lung cancer cells.Methods Experiment according to handle different were divided into A549 lung cancer A549 cell group,Lpt-7a-3p(transfection Let-7a-3p analog mimics),negative control group(negative control mimic,NC)and not transfection control(non transfected,NT)group.The Let-7a-3p level of cells in each group was detected by RT-qPCR,cell activity was detected by CCK8,oxygen consumption rate(OCR)and extracellular acidification rate(ECAR)were determined by real-time measurement of energy metabolism in hippocampal cells by XF,cell apoptosis was detected by loss cytometry,expressions of Let-7a-3p.Bcl-2,Caspase-3,Ras and AMPK were detected by Western blot.Results Let-7a-3p expression level in Let-7a-3p group was significantly up-regulated(P<0.01),while the expression level of Let-7a--3p in NC group was not significantly different from that in A549 group(P>0.05).The results of cck-8 method showed that the relative number of living cells in Let--7a-3p group was significantly decreased compared with A549 and NC group(P<0.01).The analysis results of real-time energy metabolism analyzer XF showed that the oxygen consumption(OCR)and extracellular acidification(ECAR)of A549 cells were significantly decreased after Let-7a-3p transfection compared with those of A549 and NC groups(P<0.01).The results of loss cytometry showed that the apoptosis rate of Let-7a-3p group was significantly higher than that of A549 group(P<0.01).Western blot results showed that overexpression of Let-7a-3p significantly up-regulated caspase-3 and AMPK proteins in A549 cells(P<0.01),while Bcl-2 and Ras proteins were significantly down-regulated(P<0.01).Conclusion Let-7a-3p can inhibit the activity and energy metabolism of lung cancer cell A549 and promote its apoptosis.The mechanism of Let-7a-3p mimesis on cancer cells may be to up-regu

关 键 词：Let-7a-3p AMPK 肺癌A549细胞 能量代谢 

分 类 号：R73[医药卫生—肿瘤]