人胃癌细胞中β-内酰胺酶基因不同转录本的表达特点  

Expression characteristics of different transcripts ofβ-lactamase gene in human gastric cancer cells

作　　者：聂微 刘泽莹 严芝强 成兴真 刘丽荣 杨芳 NIE Wei;LIU Zeying;YAN Zhiqiang;CHENG Xingzhen;LIU Lirong;YANG Fang(Department of Basic Clinical Laboratory and Hematology,School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Clinical Laboratory,Affiliated Cancer Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Gastrointestinal Surgery,Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学医学检验学院临床检验基础与血液学教研室,贵州贵阳550004 [2]贵州医科大学附属肿瘤医院检验科,贵州贵阳550004 [3]贵州医科大学附属医院胃肠外科,贵州贵阳550004

出　　处：《贵州医科大学学报》2022年第7期773-778,共6页Journal of Guizhou Medical University

基　　金：贵州省科技厅计划项目(19NSP015);贵州省卫生健康委科学技术基金项目(gzwjkj2021-168,gzwjkj2020-1-169)。

摘　　要：目的探讨人胃癌细胞中β-内酰胺酶基因(LACTB)不同转录本的表达特点。方法取对数期生长的人正常胃黏膜细胞株(GES-1)、不同分化程度胃癌细胞株[AGS(高分化)、MKN-28(中分化)、MKN-45(低分化)]及HGC-27(未分化)提取RNA,采用反转录PCR(RT-PCR)、巢式PCR、original TA cloning kit(T-A)克隆方法及DNA测序确定各细胞中LACTB表达的转录本类型,采用荧光定量PCR(RT-qPCR)检测LACTB不同转录本在胃癌细胞中的表达水平。结果在胃癌细胞株中检测到LACTB的3个转录本;以GES-1为对照,LACTB转录本1在AGS、MKN-28、MKN-45中高表达(P<0.05),在HGC-27中低表达(P<0.05);以GES-1为对照,转录本2在AGS、MKN-28、MKN-45、HGC-27中低表达(P<0.05),转录本3在MKN-28、MKN-45、HGC-27中低表达(P<0.05)。结论与正常胃黏膜细胞相比,胃癌细胞中LACTB转录本1表达水平增加、转录本2和3表达水平降低,转录本1可能是胃癌细胞分化的指标之一。Objective To investigate the expression characteristics ofβ-lactamase gene(LACTB)transcripts in human gastric cancer cells.Methods RNA was extracted from the human normal gastric mucosa cell lines(GES-1),gastric cancer cell lines[AGS(highly differentiated),MKN-28(moderately differentiated),MKN-45(poorly differentiated)]with different differentiation degrees,and HGC-27(undifferentiated).The transcript type of LACTB expression was determined by reverse transcript PCR(RT-PCR),nested PCR,original TA cloning kit(T-A)cloning methods,and DNA sequencing.The expression levels of the different LACTB transcripts in gastric cancer cells were determined by fluorescence quantitative PCR(RT-qPCR).Results Three transcripts of LACTB were detected in the gastric cancer cell lines.Using GES-1 as a control,LACTB transcript 1 was highly expressed in AGS,MKN-28,and MKN-45(P<0.05),and low expression was observed in HGC-27(P<0.05).Using GES-1 as a control,transcript 2 was found in low expression in AGS,MKN-28,MKN-45,and HGC-27(P<0.05),and transcript 3 was poorly expressed in MKN-28,MKN-45,and HGC-27(P<0.05).Conclusion Compared with normal gastric mucosal cells,the expression level of LACTB transcript 1 is increased in gastric cancer,and the expression levels of transcripts 2 and 3 are decreased.Transcript 1 may be one of the indicators of gastric cancer cell differentiation.

关 键 词：胃肿瘤 Β-内酰胺酶基因 可变剪接 转录本 反转录PCR 荧光定量PCR 

分 类 号：R446.9[医药卫生—诊断学]