GATA3/GATA6调控结肠癌细胞株自噬和增殖的机制  

Mechanisms of GATA3/GATA6 regulating autophagy and proliferation of colon cancer cell lines

作　　者：刘珊 刘鑫 林友刚[1] LIU Shan;LIU Xin;LIN Yougang(Department of Anorectal Surgery,Hainan General Hospital(Hainan Affiliated Hospital of Hainan medical University),Haikou 570311,China;不详)

机构地区：[1]海南省人民医院(海南医学院附属海南医院)肛肠外科,海口570311 [2]海南省人民医院(海南医学院附属海南医院)胃肠外科,海口570311

出　　处：《临床外科杂志》2022年第7期694-696,共3页Journal of Clinical Surgery

摘　　要：目的研究GATA3/GATA6调控结肠癌细胞株自噬与增殖的机制。方法以结肠癌HCT116细胞作为实验材料,建立针对GATA3及GATA6的干扰RNA表达载体及阴性对照载体,瞬时转染HCT116细胞。流式细胞术检测转染细胞周期改变,Annexin-V-FITC/PI双染检测细胞凋亡情况,应用MDC对细胞自噬进行测定。结果流式细胞检测发现,干扰GATA3/GATA6后结肠癌细胞凋亡率为(43.15±13.58)%,显著高于空白对照组的(4.13±0.46)%,差异有统计学意义(P<0.05)。对照组0小时、24小时、48小时、72小时结肠癌细胞增殖水平分别为(1.05±0.06)、(1.01±0.07)、(1.03±0.04)、(1.06±0.07),干扰GATA3/GATA6后分别为(1.04±0.04)、(0.81±0.03)、(0.61±0.06)、(0.49±0.05),结肠癌细胞存活率较空白对照组显著下降,且随时间推移呈依次下降趋势,差异均有统计学意义(P<0.05)。干扰GATA3/GATA6后结肠癌细胞内mTOR、ULK1、LC3-B mRNA表达水平分别为(0.62±0.13)、(0.51±0.06)、(0.37±0.04),较空白对照组的(0.92±0.16)、(0.86±0.13)、(0.82±0.14)显著下降,差异有统计学意义(P<0.05)。干扰GATA3/GATA6后,结肠癌细胞内mTOR、ULK1、LC3-B蛋白表达量低于空白对照组,差异有统计学意义(P<0.05)。结论干扰GATA3/GATA6能抑制结肠癌细胞自噬,进而抑制细胞增殖,增加细胞凋亡。Objective To investigate the mechanisms of GATA3/GATA6 regulating autophagy and proliferation of colon cancer cell lines.Methods Colon cancer cell line HCT116 were used as experimental materials,and transfected with the vectors expressing small interfering RNAs targeting GATA3 and GATA6 and the negative control vectors,respectively.Flow cytometry was used to detect cell cycle changes in the transfected cells,Annexin-V-FITC/PI double staining was used to detect apoptosis,and MDC staining was applied to measure cell autophagy.Results Flow cytometry showed that the apoptosis rate of colon cancer cells after GATA3/GATA6 interference[(43.15±13.58)%]was significantly higher than that of blank control group[(4.13±0.46)%,P<0.05].In the control group,the proliferation levels of colon cancer cells at 0 h,24 h,48 h and 72 h were 1.05±0.06、1.01±0.07、1.03±0.04、1.06±0.07,respectively.After GATA3/GATA6 interference,the data were 1.04±0.04、0.81±0.03、0.61±0.06、0.49±0.05,respectively.Compared with the control group,the survival rate of colon cancer cells decreased significantly,and showed a downward trend over time(P<0.05).The mRNA expression levels of mTOR,ULK1 and LC3-B in colon cancer cells after GATA3/GATA6 interference were 0.62±0.13、0.51±0.06、0.37±0.04,respectively,which were significantly decreased compared with the control group(0.92±0.16、0.86±0.13、0.82±0.14,respectively)(P<0.05).After interfering GATA3/GATA6,the protein expression levels of mTOR,ULK1 and LC3-B in colon cancer cells were significantly lower than those in control group(P<0.05).Conclusion Interfering GATA3/GATA6 may inhibit autophagy and cell proliferation in colon cancer cells,and increase apoptosis.

关 键 词：GATA3 GATA6 结肠癌 自噬 增殖 

分 类 号：R735.35[医药卫生—肿瘤]