下调清道夫受体B类成员1表达对胃癌细胞AGS增殖、迁移及侵袭的影响  

Effect of downregulating scavenger receptor class B type 1 on proliferation,migration,and invasion of gastric cancer cell line AGS

作　　者：胡鑫 杨秀兰 兰冰雪 袁婷 金泳 杜洪 程树强 韦四喜 黄海 HU Xin;YANG Xiulan;LAN Bingxue;YUAN Ting;JIN Yong;DU Hong;CHENG Shuqiang;WEI Sixi;HUANG Hai(Center for Clinical Laboratories,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Laboratory Medicine,the First Affiliated Hospital of Sun Yat-sen University Guizhou Hospital,Guiyang 550029,Guizhou,China;Department of Laboratory Medicine,Guiyang Municipal Center for Public Health Treatment,Guiyang 550004,Guizhou,China;Department of Laboratory Medicine,the Second People's Hospital of Guizhou Province,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵州贵阳550004 [2]贵州医科大学医学检验学院,贵州贵阳550004 [3]中山大学附属第一医院贵州医院检验科,贵州贵阳550029 [4]贵阳市公共卫生救治中心检验科,贵州贵阳550004 [5]贵州省第二人民医院检验科,贵州贵阳550004

出　　处：《贵州医科大学学报》2023年第12期1429-1437,1482,共10页Journal of Guizhou Medical University

基　　金：国家自然科学基金地区科学基金项目(8206100716);贵阳市科技计划项目(筑科合〔2019〕9-1-5);贵州医科大学附属医院博士科研启动基金项目(gyfybsky-2021-39)。

摘　　要：目的探讨清道夫受体B类成员1(SCARB1)对胃癌细胞(AGS)增殖、迁移及侵袭的影响及其机制。方法采用RT-qPCR和Western blot检测SCARB1在人胃黏膜上皮细胞GES1和人胃癌细胞AGS中的表达水平;将AGS细胞随机分为空白对照组、阴性对照组和实验组,在实验组中使用RNA干扰技术下调SCARB1表达,阴性对照组使用siRNA阴性序列进行转染,空白对照组不转染siRNA;克隆形成实验和细胞计数试剂盒-8(CCK-8)法评估各组细胞增殖能力,流式细胞术检测各组细胞凋亡情况,细胞划痕实验和Transwell实验分析各组细胞横向迁移、纵向迁移和侵袭能力,Western blot检测基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)及PI3K/AKT通路相关蛋白的表达。结果与GES1细胞比较,SCARB1在AGS中表达升高(P<0.05),AGS细胞SCARB1干扰效率为(68.06±2.04)%;实验组细胞克隆形成率下降,在转染24 h、48 h及72 h时细胞活力均低于阴性对照组,实验组细胞凋亡率高于阴性对照组(P<0.05),阴性对照组与空白对照组比较,差异无统计学意义;实验组在24 h和48 h的平均划痕宽度大于阴性对照组,48 h时细胞迁移和侵袭数量少于阴性对照组(P<0.05),阴性对照组与空白对照组比较,差异无统计学意义;实验组MMP2、MMP9、AKT、p-AKT(ser473)、PI3K(p110)表达下降,p-AKT/AKT减少(P<0.05),阴性对照组与空白对照组比较,差异无统计学意义。结论SCARB1在AGS中高表达,下调SCARB1表达作用于PI3K/AKT信号通路和MMP2、MMP9蛋白而抑制胃癌细胞AGS增殖、迁移和侵袭,促进其凋亡。Objective To investigate the effect of scavenger receptor class B type 1(SCARB1)on the proliferation,migration and invasion of gastric cancer cell line AGS and its mechanism.Methods RT-qPCR and Western blot were applied to detect the expression level of SCARB1 in human gastric mucosal epithelial cell line GES1 and human gastric cancer cell line AGS.AGS cells were randomly divided into blank control group,negative control group and experimental group.Cells in experimental group were transfected with siRNA against SCARB1 to silence SCARB1 expression by RNA interference,while cells in negative group were transfected with siRNA negative control sequence.Blank control group was not transfected with siRNA.Colony formation assay and cell counting kit 8(CCK-8)assay were used to evaluate the proliferation ability of each group.Flow cytometry was used to detect cellular apoptosis of each group.Cell scratch assay and Transwell assay were used to analyze the lateral migration,longitudinal migration and invasion capacity of each group.Western blot was performed to detect the expressions of matrix metalloproteinase 2(MMP2),MMP9,and PI3K/AKT pathway-related proteins.Results When compared with GES1 cells,SCARB1 expression was increased in AGS(P<0.05).The interference efficiency of SCARB1 in AGS cells was(68.06±2.04)%.Colony formation rate was decreased in experiment group.At 24,48,and 72 hours of transfection,the cell viability was lower in experiment group than that of negative control group,and the apoptosis rate of experimental group was higher than that of negative control group(P<0.05).There was no statistically significant difference in apoptosis rate between negative control and blank control groups.The average scratch widths of experimental group were greater than those of negative control group at 24 and 48 hours,while the numbers of cell migration and invasion were less than those of negative control group at 48 hours(P<0.05).There were no statistically significant differences between negative control and blank contro

关 键 词：清道夫受体B类成员1 胃肿瘤 人胃腺癌细胞 增殖 迁移 侵袭 磷脂酰肌醇3激酶 

分 类 号：R735.2[医药卫生—肿瘤]