微小RNA-3679与小鼠异种移植物肿瘤生长的关系研究  

AmiRNA-3679 Promotes Tumor Growth in Mouse Xenografts

作　　者：何静宇 周雪琴 李艺明[1] 王文涛[2] He Jingyu;Zhou Xueqin;Li Yiming;Wang Wentao(Department of Hepatobiliary and Pancreatic Surgery,The Third Hospital of Mianyang•Sichuan Mental Health Center,Mianyang 621000,China;Department of Liver Surgery,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]绵阳市第三人民医院•四川省精神卫生中心肝胆胰外科,绵阳621000 [2]四川大学华西医院肝脏外科,成都610041

出　　处：《成都医学院学报》2023年第5期555-559,564,共6页Journal of Chengdu Medical College

基　　金：绵阳市卫生健康委员会鼓励科研项目(No:202216)。

摘　　要：目的探讨微小RNA(miRNA)-3679对小鼠异种移植物(Hep3B)肿瘤生长影响的机制研究。方法将裸鼠随机分为3组,空白对照组(NC组),抑制miRNA-3679组(CT1组)及同时转染miRNA-3679 inhibitor与si-ZADH2组(CT2组);裸鼠成瘤实验检测各组瘤体质量及体积;实时荧光定量PCR(qPCR)检测相关基因表达;蛋白质免疫印迹检测瘤组织中靶蛋白及相关凋亡蛋白;免疫组化染色测定Caspase-3的表达;Ki-67免疫组织化学染色分析细胞增殖情况。结果28 d,CT1组肿瘤体积及质量均小于NC组(P<0.01);CT1组及CT2组中miRNA-3679的表达低于NC组(P<0.01),CT1组中的ZADH2表达高于NC组及CT2组(P<0.001);蛋白质免疫印迹检测提示,CT1组中的ZADH2蛋白表达升高(P<0.001),CT2组ZADH2蛋白表达低于CT1组(P<0.001);CT1组Bax、Caspase-3、Caspase-9蛋白表达高于NC组和CT2组(P<0.05),但CT1组Bcl-2蛋白表达低于NC组和CT2组(P<0.05);免疫组织化学染色显示,CT1组Caspase-3的表达高于NC组和CT2组(P<0.05);Ki-67免疫组织化学染色提示,CT1组肿瘤细胞数量少于NC组和CT2组(P<0.01)。结论miRNA-3679可促进小鼠异种移植物(Hep3B)肿瘤的发生与发展,且miRNA-3679可通过靶基因ZADH2发挥作用。Objective To study the mechanism of miRNA-3679 promoting tumor growth of xenograft(Hep3B)in mice.Methods Nude mice were randomly divided into three groups:blank control(NC)group,miRNA-3679 inhibitor(CT1)group and simultaneous transfection of miRNA-3679 inhibitor and si-ZADH2(CT2)group.Nude mouse tumorigenesis experiment was used to detect the mass and volume of tumor in each group.The expression of related genes was detected by quantitative polymerase chain reaction(qPCR).Western blot was used to detect the target proteins and apoptosis related proteins in tumor tissues.And immunohistochemical staining was used to detect the expression of Caspase-3.Ki-67 immunohistochemical staining was used to analyze the cell proliferation.Results On the 28th day,the tumor volume and mass in CT1 group were smaller than those in NC group(P<0.01);The expression of miRNA-3679 in CT1 group and CT2 group was lower than that in NC group(P<0.01);And the expression of ZADH2 in CT1 group was higher than that in NC group and CT2 group(P<0.001);Western blot showed that the expression of ZADH2 protein in CT1 group was increased(P<0.001),and that in CT2 group was lower than that in CT1 group(P<0.001);The protein expressions of Bax,Caspase-3 and Caspase-9 in CT1 group were higher than those in NC group and CT2 group(P<0.05),while the protein expression of Bcl-2 in CT1 group was lower than that in NC group and CT2 group(P<0.05);Immunohistochemical staining showed that the expression of Caspase-3 in CT1 group was higher than that in NC group and CT2 group(P<0.05);Ki-67 immunohistochemical staining showed that the number of tumor cells in CT1 group was less than that in NC group and CT2 group(P<0.01).Conclusion miRNA-3679 can promote the occurrence and development of xenograft(Hep3B)tumor in mice.And miRNA-3679 can play a role through the target gene ZADH2.

关 键 词：微小RNA 微小RNA-3679 含锌结合醇脱氢酶结构域-2 可塑性相关基因蛋白-3 

分 类 号：R394.3[医药卫生—医学遗传学]