三结构域家族蛋白22在宫颈癌中的表达及其对高危型人乳头瘤病毒阳性宫颈癌细胞增殖、侵袭及凋亡的影响  

Expression of Tripartite Motif Protein 22 in Cervical Cancer and Its Influence on Proliferation,Invasion and Apoptosis of Cervical Cancer Cells with High Risk Human Papillomavirus Infection

作　　者：王海鹏[1] 刘屹[1] 陈庆芬 王希方 侯银银 WANG Hai-peng;LIU Yi;CHEN Qing-fen;WANG Xi-fang;HOU Yin-yin(Shaanxi Provincial People's Hospital,Xi'an 710068,China;The Fourth People's Hospital of Shaanxi,Xi'an 710043,China)

机构地区：[1]陕西省人民医院,陕西西安710068 [2]陕西省第四人民医院,陕西西安710043

出　　处：《肿瘤学杂志》2021年第9期758-765,共8页Journal of Chinese Oncology

基　　金：国家自然科学基金(81402012);陕西省自然科学研究基金(2015JQ8321,2019JM-547);西安交通大学基本业务项目(xyz012019112);西安市科技计划项目[2019114613YX001SF035(3)]。

摘　　要：[目的]探讨三结构域家族蛋白22(tripartite motif protein 22,TRIM22)在宫颈癌中的表达及其与临床病理学特征的关系,并分析TRIM22对高危型人乳头瘤病毒(human papilloma virus,HPV)阳性宫颈癌细胞增殖、侵袭和凋亡的影响。[方法]收集2017年8月至2018年9月41例手术切除或活检的宫颈癌组织和18例正常子宫颈组织,分别提取总RNA和总蛋白。实时定量PCR分析TRIM22 mRNA表达,Western blot分析TRIM22蛋白水平,并分析TRIM22 mRNA表达变化与患者临床病理学特征的关系。以HPV16阳性宫颈癌细胞系CaSki和HPV18阳性宫颈癌细胞系HeLa为研究对象,用TRIM22表达质粒(pUNO1-hTRIM22a)和对照表达质粒(p UNO1)稳定转染Ca Ski细胞和HeLa细胞,以未转染细胞为对照组。实时定量PCR和Western blot检测转染后细胞中TRIM22 mRNA表达和蛋白水平。采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)检测细胞增殖;Transwell细胞侵袭实验检测细胞侵袭能力;流式细胞术检测细胞周期和凋亡;Western blot检测抗磷脂酰肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(protein kinase B,AKT)和磷酸化AKT(phosphorylated AKT,p-AKT)蛋白表达。[结果]TRIM22在宫颈癌组织中mRNA表达(0.99±0.16 vs 4.39±1.41,t=15.40,P<0.001)和蛋白水平(0.30±0.12 vs 1.32±0.27,t=19.97,P<0.001)均明显低于正常子宫颈组织,TRIM22在高危型HPV阳性宫颈癌组织中mRNA表达(0.94±0.14 vs 1.17±0.12,t=4.21,P<0.001)和蛋白水平(0.27±0.08 vs 0.43±0.18,t=3.86,P<0.001)均明显低于高危型HPV阴性宫颈癌组织。TRIM22 mRNA表达与淋巴结转移明显相关(1.04±0.16 vs 0.89±0.13,t=3.15,P=0.003),与肿瘤大小、临床分期无明显相关(P>0.05)。pUNO1转染CaSki细胞和HeLa细胞与未转染细胞在TRIM22 mRNA和蛋白水平、细胞增殖、侵袭数、细胞周期、凋亡率、PI3K、AKT、pAKT蛋白表达量的差异均无统计学意义(P>0.05),pUNO1-hTRIM22a转染后CaSki细胞和HeLa细胞中TRIM22mRNA和蛋白水平显著性�[Objective]To investigate the expression of tripartite motif protein 22(TRIM22)and its effects on proliferation,invasion and apoptosis of cervical cancer cells infected with high risk human papilloma virus(HPV).[Methods]Forty-one cervical cancer tissue samples and 18 normal cervical tissue samples were collected between August 2017 and September 2018.Total RNA and protein in tissue samples were extracted respectively.TRIM22 m RNA expression was analyzed by real-time RT-PCR,while TRIM22 protein level was analyzed by Western blot.The relationship between TRIM22 m RNA expression and clinical pathological features of patients was analyzed.The HPV16-positive cervical cancer Ca Ski cells and HPV18-positive cervical cancer He La cells were transfected with TRIM22 expression plasmid(p UNO1-h TRIM22 a)and control plasmid(p UNO1),the corresponding untransfected cells were used as control group.TRIM22 m RNA and protein expression was analyzed by q RT-PCR and Western blot,respectively.Cell proliferation was measured by cell counting kit-8(CCK-8),cell invasion ability was detected by Transwell assay,cell cycle and apoptosis was determined by flow cytometry,the expression of phosphatidylinositol3-kinase(PI3 K),protein kinase B(AKT)and phosphorylated AKT(p-AKT)was detected by Western blot.[Results]TRIM22 m RNA(0.99±0.16 vs 4.39±1.41,t=15.40,P<0.001)and protein level(0.30±0.12 vs 1.32±0.27,t=19.97,P<0.001)in cervical cancer tissues was significantly lower than those in normal cervical tissues.TRIM22 m RNA(0.94±0.14 vs 1.17±0.12,t=4.21,P=0.001)and protein level(0.27±0.08 vs 0.43±0.18,t=3.86,P<0.001)in high risk HPV-positive cervical cancer tissues was significantly lower than that in high risk HPV-negative cervical cancer tissues.TRIM22 m RNA expression was correlated with lymph node metastasis(1.04±0.16 vs 0.89±0.13,t=3.15,P=0.003),however,did not correlate with tumor size or clinical stage(P>0.05).After transfection,TRIM22 m RNA and protein level was remarkably elevated in p UNO1-h TRIM22 a group(P<0.001).Cell prolife

关 键 词：宫颈肿瘤 乳头状瘤病毒 三结构域家族蛋白 细胞增殖 细胞凋亡 侵袭能力 

分 类 号：R737.33[医药卫生—肿瘤]