沉默GTPBP4基因表达对食管癌细胞增殖、侵袭和化疗敏感性影响  

Effects of GTPBP4 Gene Silencing on Proliferation,Invasion and Chemosensitivity of Human Esophageal Carcinoma Cells

作　　者：张翠红 吕欣 侯春立 张建军 马博敬 范才 Zhang Cuihong;Lv Xin;Hou Chunli;Zhang Jianjun;Ma Bojing;Fan Cai(Department of Radiation Oncology,the 9S0th Hospital of PLA Joint Logistics Support Force,Shijiazhuang 050082,Hebei,China;Department of Oncology,the 9S0th Hospital of PLA Joint Logistics Support Force,Shijiazhuang 050082,Hebei,China;Department of Medical Services,Central Hospital of Baixiang County,Xingtai 055450,Hebei,China)

机构地区：[1]解放军联勤保障部队第九八〇医院放疗科,石家庄050082 [2]柏乡县中心医院医务科,河北邢台055450 [3]解放军联勤保障部队第九八〇医院肿瘤科,石家庄050082

出　　处：《肿瘤预防与治疗》2021年第3期190-198,共9页Journal of Cancer Control And Treatment

基　　金：河北省卫生厅青年科技课题(编号:20150382)。

摘　　要：目的:探讨沉默GTPBP4基因表达对人食管癌EC109细胞生长、侵袭及化疗药物敏感性的影响.方法:通过GEO数据库,分析食管癌患者组织中GTPBP4 mRNA的表达水平;利用慢病毒(LV-GTPBP4-siRNA组)及阴性病毒(LV-NC组)转染食管癌EC109细胞后,采用实时荧光定量PCR(real-time quantitative polymerase chain reaction,RT-qPCR)和蛋白质印迹法(Western blot)检测GTPBP4基因表达沉默效果,然后采用CCK8法、流式细胞法、Transwell小室实验分别检测GTPBP4基因表达沉默后EC109细胞增殖、周期分布、侵袭能力及化疗敏感性变化.结果:GEO数据库分析表明GTPBP4 mRNA在人食管癌组织中的表达明显高于正常邻近食管组织(P<0.001).EC9706、EC109和KYSE-150细胞GTPBP4 mRNA相对表达水平分别为2.66±0.32、4.62±0.06、3.38±0.17,明显高于HEEC细胞(均P<0.001);EC9706、EC109和KYSE-150细胞GTPBP4蛋白相对表达水平分别为2.46±0.56、3.95±0.36、3.18±0.10,明显高于HEEC细胞(P<0.01,P<0.001,P<0.001).LV-GTPBP4-siRNA病毒转染后EC109细胞中GT-PBP4 mRNA及蛋白表达水平明显下降(P<0.05,P<0.001).GTPBP4基因表达沉默后,EC109细胞的生长受到明显抑制(P<0.001),且加入不同浓度顺铂和5-氟尿嘧啶处理细胞后,细胞的存活率和药物半数抑制浓度(50%in-hibiting concentration,IC50)均降低(均P<0.05);同时G0/G1期细胞所占比例明显升高,S期细胞所占比例降低(均P<0.001);并且GTPBP4基因表达沉默及联合化疗药物顺铂处理使EC109细胞的穿膜细胞数量明显减少,侵袭能力降低(P<0.001,P=0.001).结论:人食管癌组织中GTPBP4 mRNA的表达明显高于正常邻近食管组织.LV-GTPBP4-siRNA慢病毒载体可有效抑制食管癌EC109细胞中GTPBP4基因的表达,从而抑制细胞增殖及侵袭能力,增强人食管鳞癌细胞对化疗药物的敏感性.Objective: To investigate the effects of GTPBP4 gene silencing on growth, invasion and and chemosensitivity of human esophageal carcinoma cells. Methods: The expression of GTPBP4 mRNA in esophageal cancer was analyzed using Bioinformatics analysis of microarray data in GEO database. The effect of GTPBP4 gene silencing was detected by real-time quantitative polymerase chain reaction(RT-qPCR) and Western blot after esophageal carcinoma cells were transfected with lentivirus(the LV-GTPBP4-siRNA group) and negative virus(the LV-NC group), respectively. The changes in proliferation ability, cell cycle distribution, invasion and chemosensitivity of esophageal carcinoma EC109 cells after GTPBP4 gene silencing were detected by CCK8, flow cytometry(FCM) assay and Transwell invasion assay, respectively. Results: The expression level of GTPBP4 mRNA in esophageal cancer tissues were significantly higher than that in adjacent normal esophageal tissues(P<0.001). The relative expression levels of GTPBP4 mRNA in EC9706, EC109 and KYSE-150 cells were 2.66±0.32, 4.62±0.06 and 3.38±0.17, respectively, which were significantly higher than those in human esophageal epithelial cells(HEEC)(all P<0.001);the relative expression levels of GTPBP4 protein in EC9706, EC109 and KYSE-150 cells were 2.46±0.56, 3.95±0.36 and 3.18±0.10, respectively, which were significantly higher than those in HEEC(P<0.01, P<0.001, P<0.001). The expressions of GTPBP4 mRNA and GTPBP4 protein in EC109 cells significantly decreased after GTPBP4 gene silencing(P<0.05, P<0.001). After GTPBP4 gene silencing, the proliferation ability of EC109 cells significantly decreased, the proportion of cells in G0/G1 phase significantly increased, and the proportion of cells in S phase significantly decreased(all P<0.001);and survival rates and 50% inhibiting concentration(IC50) of EC109 cells both significantly decreased after different concentrations of cisplatin or 5-FU were further added(all P<0.05). Meanwhile, the number of invasion cells significantly decreased both

关 键 词：食管癌 人类GTP结合蛋白4 RNA干扰 慢病毒 增殖 化疗敏感性 

分 类 号：R735.1[医药卫生—肿瘤] R730.231[医药卫生—临床医学]