组蛋白去甲基化酶JMJD2D对人胃癌AGS细胞恶性表型的影响  

Effects of histone demethylase JMJD2D on the cellular malignant phenotype of human gastric cancer AGS cells

作　　者：余齐鸣 洪莲莲 胡旋宇 李沛[2] 凌志强 Yu Qiming;Hong Lianlian;Hu Xuanyu;Li Pei;Ling Zhiqiang(Experimental Research Centre,Cancer Hospital of University of Chinese Academy of Sciences(Zhejiang Cancer Hospital),Institute of Cancer and Basic Medicine,Chinese Academy of Sciences,Hangzhou 310022,China;Department of Pathophysiology,School of Basic Medical Sciences,Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]中国科学院大学附属肿瘤医院(浙江省肿瘤医院)实验研究中心中国科学院肿瘤与基础医学研究所,杭州310022 [2]郑州大学基础医学院病理生理学教研室,450052

出　　处：《中华实验外科杂志》2021年第3期425-428,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金面上项目(81372332、81572822、81972908);浙江省自然科学基金重点项目(LZ13H160002、LZ18H160002);浙江省自然科学基金面上项目(LY17H160044、LQ19H160005);浙江省"万人计划"科技创新领军人才(浙委人[2019]3号);浙江省卫生高层次创新人才培养工程基金项目(浙卫发[2014]108号);浙江省新世纪151人才工程重点层次项目(浙人社发[2014]150号)。

摘　　要：目的观察组蛋白去甲基化酶JMJD2D在人胃癌细胞系AGS中表达下调后对细胞生物学功能的影响。方法采用小发夹状RNA(small hairoin RNA,shRNA)技术干扰AGS细胞中JMJD2D表达,通过反转录-聚合酶链反应(RT-PCR)检测JMJD2D沉默效果,同时设置对照组;利用细胞计数试剂盒(CCK-8)法检测JMJD2D对AGS细胞增殖能力的影响;利用划痕实验Transwell侵袭实验检测JMJD2D对AGS细胞迁移能力的影响;利用流式细胞技术检测JMJD2D对AGS细胞周期和凋亡的影响。组间比较采用单因素方差分析,进一步比较行配对t检验。结果通过shRNA技术成功下调AGS细胞中JMJD2D表达,细胞增殖实验结果显示,在培养24、48、72 h后,JMJD2D shRNA组细胞的增殖水平(0.311±0.012、0.475±0.038、0.813±0.043)明显低于对照组(0.385±0.013、0.558±0.042、0.917±0.035),差异均有统计学意义(t=3.960、5.370、7.830,P<0.01);划痕实验结果表明,划痕24 h后,JMJD2D shRNA组细胞的相对倍数(0.53±0.09)明显低于对照组(0.99±0.03),差异有统计学意义(t=4.940,P<0.01);Transwell迁移实验结果表明,JMJD2D shRNA组迁移到膜下的细胞数[(155.0±10.6)个]明显低于对照组[(312.0±18.3)个],差异有统计学意义(t=7.530,P<0.01);细胞周期检测结果显示,JMJD2D shRNA组细胞阻滞于G1期(t=4.290,P<0.05);细胞凋亡试验显示,抑制JMJD2D表达可明显增加AGS细胞凋亡(JMJD2D shRNA组细胞凋亡率18.5%,对照组3.1%),差异有统计学意义(t=9.270,P<0.01)。结论下调JMJD2D表达可明显抑制AGS细胞增殖及迁移能力,促进细胞凋亡,提示JMJD2D在胃癌细胞中起着癌基因作用。Objective To investigate the effects of histone demethylase JMJD2D on the cellular malignant phenotype of human gastric cancer AGS cells.Methods The expression of JMJD2D in AGS cells was interfered with small hair RNA(shRNA),and the expression level of JMJD2D was detected by real-time RT-PCR.The control group was set up at the same time.We employed cell counting kit-8(CCK-8)assay to investigate the effects of JMJD2D on the proliferation of AGS cells.Wound healing assay and Transwell assay were used to test the effects of JMJD2D on the migration of AGS cells.Flow cytometry was used to monitor the effects of JMJD2D on cell cycle and cell apoptosis of AGS cells.Results The expression of JMJD2D in AGS cells was successfully downregulated by shRNA technique.The relative proliferation rate of JMJD2D shRNA AGS cells cultured for 24,48 and 72 h was significantly lower than that in the control cells(0.311±0.012 vs.0.385±0.013,t=3.960,P<0.01;0.475±0.038 vs.0.558±0.042,t=5.370,P<0.01;0.813±0.043 vs.0.917±0.035,t=7.830,P<0.01).The relative migration rate of JMJD2D shRNA AGS cells at 24 h after scratching was significantly lower than that of control cells(0.53±0.088 vs.0.99±0.031,t=4.940,P<0.01).Transwell assay showed that the number of migrating cells in the JMJD2D shRNA group was significantly less than that in the control group(155.0±10.6 vs.312.0±18.3,t=7.530,P<0.01).The results of flow cytometry showed that JMJD2D shRNA AGS cells were blocked at G1 phase(t=4.290,P<0.05),and that the inhibition of JMJD2D could significantly influence the apoptosis of AGS cells(18.5%vs.3.1%,t=9.270,P<0.01).Conclusion The down-regulation of JMJD2D expression can significantly inhibit the proliferation and migration of AGS cells and promote apoptosis,suggesting that JMJD2D plays an oncogenic role in gastric cancer cells.

关 键 词：胃癌 增殖 迁移 细胞周期 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]