下调ACTG1抑制PI3K/Akt信号通路相关蛋白表达促进胃癌细胞HGC-27凋亡机制研究  

Mechanism of ACTG1 Downregulation Inhibits the Expression of Protein Associated to PI3K/Akt Signaling Pathway and Promotes Gastric Cancer Cells Line HGC-27 Apoptosis

作　　者：李寿敏 兰冰雪 潘莉 孙铭 崔淼淼 韦四喜 黄海 LI Shoumin;LAN Bingxue;PAN Li;SUN Ming;CUI Miaomiao;WEI Sixi;HUANG Hai(Center for Clinical Laboratories,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,China;School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,China;Department of Clinical Laboratory,Liupanshui Municipal People’s Hospital,Guizhou Liupanhui 553001,China;Department of Clinical Laboratory,Guiyang Nanming District People’s Hospital,Guiyang 550004,China;Department of Clinical Laboratory,Guiyang Second People’s Hospital,Guiyang 550004,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵阳550004 [2]贵州医科大学医学检验学院,贵阳550004 [3]六盘水市人民医院检验科,贵州六盘水553001 [4]贵阳市南明区人民医院检验科,贵阳550004 [5]贵阳市第二人民医院检验科,贵阳550004

出　　处：《现代检验医学杂志》2025年第2期1-5,共5页Journal of Modern Laboratory Medicine

基　　金：国家自然科学基金(82060442);2021年贵州医科大学附属医院博士科研启动基金(gyfybsky-2021-39);贵州医科大学国家自然科学基金培育项目(21NSFCP05);2021年贵州医科大学附属医院国家自然科学基金培育项目(gyfynsfc-2021-11)。

摘　　要：目的探讨下调γ-肌动蛋白(ACTG1)对人胃癌细胞(未分化)(HGC-27)凋亡的影响及机制。方法利用CRISPR/Cas9技术构建稳定敲低ACTG1 HGC-27细胞系;通过DNA测序及Western blot验证ACTG1敲低情况;流式细胞术检测细胞凋亡率;Western blot检测凋亡相关蛋白Bcl2相关X蛋白(Bax)、B细胞淋巴瘤2家族蛋白(Bcl2)以及PI3K(磷脂酰肌醇3-激酶)/AKT(蛋白激酶B)通路相关蛋白AKT,p-AKT的表达水平。结果构建稳定敲低ACTG1的HGC-27细胞系。与Control组相比,ACTG1蛋白水平在sgACTG1-1组和sgACTG1-2组显著下降。与Control组细胞凋亡率相比,sgACTG1-1组和sgACTG1-2组细胞凋亡率(10.11%±0.46%,14.67%±0.17%vs 6.54%±0.67%)明显升高,差异具有统计学意义(t=-7.58,-20.28,均P<0.01);与Control组相比,sgACTG1-1组和sgACTG1-2组Bax蛋白水平(0.89±0.02,1.00±0.08 vs 0.71±0.03)显著升高(t=-8.14,-5.87),Bcl2蛋白水平(0.49±0.06,0.39±0.06 vs 0.65±0.07)显著下降(t=3.09,5.35),差异具有统计学意义(均P<0.05)。与Control组相比,sgACTG1-1组和sgACTG1-2组AKT蛋白水平(0.95±0.10,0.43±0.09 vs 1.17±0.06)和p-AKT蛋白水平(0.38±0.08,0.28±0.12 vs 0.70±0.14)显著下降,差异具有统计学意义(t=3.20,12.13;3.44,3.85,均P<0.05)。结论下调ACTG1抑制PI3K/Akt信号通路相关蛋白AKT,p-AKT的表达,促进胃癌细胞HGC-27凋亡。Objective To investigate the effect and mechanism of actin gamma1(ACTG1)downregulation on apoptosis in gastric cancer cells line HGC-27.Methods Stable ACTG1 knockdown HGC-27 cell line was constructed by CRISPR/Cas9,ACTG1 knockdown was verified by DNA sequencing and Western blot,apoptosis rate was detected by flow cytometry,Western blot detected the expression level of apoptosis-related proteins Bcl2-associated X protein(Bax),B cell lymphoma 2 family protein(Bcl2)and PI3K/AKT signal pathway-related proteins AKT and p-AKT.Results HGC-27 cell lines with stable knockdown of ACTG1 were constructed,and the ACTG1 protein levels were decreased in the sgACTG1-1 and sgACTG1-2 groups compared with the Control group.Compared with the apoptosis rate in the Control group(6.54%±0.67%),cell apoptosis rate in the sgACTG1-1 and sgACTG1-2 groups(10.11%±0.46%,14.67%±0.17%)were significantly increased,the differences were statistically significant(t=-7.58,-20.28,all P<0.01).Compared the Control group,the Bax protein levels in the sgACTG1-1 and sgACTG1-2 groups(0.89±0.02,1.00±0.08 vs 0.71±0.03)were significantly increased(t=-8.14,-5.87),the level of Bcl2 protein(0.49±0.06,0.39±0.06 vs 0.65±0.07)were significantly decreased(t=3.09,5.35),the differences were statistically significant(all P<0.05).Compared with the Control group,the AKT protein levels in the sgACTG1-1 and sgACTG1-2 groups(0.95±0.10,0.43±0.09 vs 1.17±0.06)and the P-AKT protein level(0.38±0.08,0.28±0.12 vs 0.70±0.14)were significantly decreased,the differences were statistically significant(t=3.20,12.13;3.44,3.85,all P<0.05).Conclusion Downregulation of ACTG1 inhibits the expression of PI3K/Akt signaling pathway related proteins AKT,p-AKT and promotes gastric cancer cells line HGC-27 apoptosis.

关 键 词：γ-肌动蛋白 人胃癌细胞(未分化) 磷脂酰肌醇3-激酶/蛋白激酶B 凋亡 

分 类 号：R735.2[医药卫生—肿瘤] R730.43[医药卫生—临床医学]