胃癌细胞中Cullin1基因表达与肿瘤细胞凋亡的关系  

Relationship between Cullin1 gene expression and tumor cell apoptosis in gastric cancer cells

作　　者：杨沛刚[1] 田园[1] 檀碧波[1] 丁平安 李勇[1] 赵群[1] 陈祖建 YANG Peigang;TIAN Yuan;TAN Bibo;DING Ping'an;LI Yong;ZHAO Qun;CHEN Zujian(Department of Surgery,the Fourth Hospital of Hebei Medical University,Hebei Shijiazhuang 050011,China;Linxi County People's Hospital,Hebei Xingtai 054900,China)

机构地区：[1]河北医科大学第四医院外三科,河北石家庄050011 [2]河北省临西县人民医院,河北邢台054900

出　　处：《现代肿瘤医学》2022年第14期2487-2491,共5页Journal of Modern Oncology

基　　金：河北省医学科学研究重点课题计划(编号:20180547);河北省高等学校科学技术研究项目(编号:ZD2019139)。

摘　　要：目的:探讨胃癌组织及细胞中Cullin1基因表达水平及其对凋亡的影响及其机制。方法:在TCGA(The Cancer Genome Atlas)数据库下载胃癌数据,分析胃癌组织与正常组织中Cullin1表达差异。应用实时荧光定量PCR(qRT-PCR)技术检测36例胃癌及正常组织中Cullin1基因表达。合成小干扰RNA(Cullin1-siRNA)转染胃癌细胞系AGS,抑制Cullin1表达;四甲基偶氮唑盐比色法(MTT)检测细胞的增殖活性;流式细胞仪实验检测细胞凋亡率;qRT-PCR和蛋白印迹(Western blot)检测转染各组细胞Cullin1、Bcl2、Bax和Survivin、Livin基因mRNA和蛋白表达。结果:生物信息学结果显示,Cullin1基因在胃癌组织中表达水平明显高于正常组织(P<0.01);不同T分期的胃癌组织中Cullin1基因表达存在差异(P=0.026)。qRT-PCR结果显示验证结果与生物信息学结果符合。Cullin1-siRNA能有效沉默AGS细胞Cullin1基因的表达;有效抑制AGS细胞Cullin1表达后,转染组细胞的活性明显低于NS-siRNA组和空白对照组;凋亡率在Cullin1-siRNA组明显高于NS-siRNA组、空白对照组。Cullin1-siRNA转染后AGS中Cullin1和Bcl2、Survivin、Livin基因和蛋白表达下调,而Bax基因和蛋白表达上调(P<0.05)。结论:Cullin1基因在胃癌中表达增强且与肿瘤浸润深度有关,该基因可能通过抑制胃癌细胞凋亡而发挥作用。Objective:To explore the expression level of Cullin1 gene in gastric cancer tissues and cells and its effect on apoptosis and its mechanism.Methods:Data of gastric cancer was collected from The Cancer Genome Atlas(TCGA)database,and the difference about Cullin1 expression between gastric cancer tissues and normal tissues was analyzed.The Quantitative Real Time PCR(qRT-PCR)technique was used to detect the expression of Cullin1 gene in gastric cancer and normal tissues from 36 patients.Small interfering RNA(Cullin1-siRNA)was transfected into GASTRIC cancer cell line AGS to inhibit Cullin1 expression.MTT assay was used to detect the proliferative activity of cells.Flow cytometry was used to detect the apoptosis rate.The mRNA and protein expressions of Cullin1,Bcl2,Bax,Survivin and Livin in the transfected cells were detected by qRT-PCR and Western blot.Results:Bioinformatics results showed that the expression level of Cullin1 gene in gastric cancer tissues was significantly higher than that in normal tissues(P<0.01).There were differences in the expression of Cullin1 gene in gastric cancer tissues of different T stages(P=0.026).The qRT-PCR results showed that the verification results were consistent with the bioinformatics results.Cullin1-siRNA can effectively silence the expression of Cullin1 gene in AGS cells.After Cullin1 expression of AGS cells was effectively inhibited,the activity of transfected cells was significantly lower than that of NS-siRNA group and blank control group.The apoptosis rate of Cullin1-siRNA group was significantly higher than NS-siRNA group and blank control group.The expressions of Cullin1,Bcl2,Survivin and Livin genes and proteins were down-regulated in AGS after cullin1-siRNA transfection,while the expressions of Bax genes and proteins were up-regulated in AGS(P<0.05).Conclusion:The expression of Cullin1 gene is enhanced in gastric cancer and is related to the depth of tumor invasion,which may play a role by inhibiting the apoptosis of gastric cancer cells.

关 键 词：胃癌 Cullin1 基因干扰 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]