miR-1197负调控Smad3抑制结肠癌细胞SW620的增殖、侵袭、迁移能力及S期细胞周期的聚集相关研究  

Studies on the negative regulation of mir-1197 to inhibit the proliferation,invasion,migration and the ability of Smad3 to inhibit SW620 cells in colon cancer

作　　者：张隆陶[1] 王劲[1] 张贻庆[1] 崔贵医 ZHANG Long-tao;WANG Jin;ZHANG Yi-qing(Department of Hepatobiliary And Pancreatic Surgery,The People’s Hospital of Jiaozuo City,Jiaozuo 454000,China)

机构地区：[1]焦作市人民医院肝胆胰疝外科,河南焦作454000

出　　处：《中国实验诊断学》2019年第5期874-879,共6页Chinese Journal of Laboratory Diagnosis

摘　　要：目的研究miR-1197负调控Smad3抑制结肠癌细胞SW620的增殖、侵袭、迁移能力及S期细胞周期的聚集,分析miR-1197在结肠癌发生及发展中的作用。方法 qRT-PCR检测miR-1197在结肠癌患者血清,肿瘤组织和癌旁正常组织中的表达水平并分析其与结肠癌患者临床特征的相关性;采用miRBase筛选miR-1197的潜在靶基因Smad3并通过荧光素酶报告基因法,qRT-PCR及Western blot验证miR-1197对Smad3的靶向调控作用;Western blot检测结肠癌肿瘤组织和癌旁正常组织中Smad3的表达量;采用CCK8法,流式细胞法,Tranwell侵袭实验,细胞划痕实验分别检测miR-1197与Smad3对SW620细胞的增殖,周期,侵袭和迁移能力的影响;构建结肠癌裸鼠移植瘤模型,检测miR-1197在裸鼠体内对肿瘤生长及细胞增殖的影响。结果血清中miR-1197的表达水平与结肠癌相关临床指标呈负相关;miR-1197在结肠癌肿瘤组织中的表达水平显著低于癌旁正常组织,在发生远端转移肿瘤组织中的表达水平显著低于未转移肿瘤组织;miR-1197通过结合Smad3的3’-UTR靶向调控Smad3的mRNA及蛋白表达;Smad3在肿瘤组织中的表达水平显著高于癌旁正常组织;miR-1197抑制SW620细胞的增殖、侵袭、迁移及S期细胞周期的聚集,但这种抑制效果可被Smad3逆转;miR-1197可抑制裸鼠移植瘤的生长及细胞增殖能力。结论 miR-1197通过负调控Smad3的表达在结肠癌中发挥抑癌作用,具有成为结肠癌诊断与预测新靶点的潜力。Objective to investigate the inhibitory effect of miR-1197 on the proliferation,invasion,migration and S phase cell aggregation of colon cancer cell SW620 via Smad3,and to analyze the role of miR-1197 in the occurrence and development of colon cancer.Methods qRT-PCR was used to detect the expression of miR-1197 in the serum,tumor tissue and adjacent normal tissue of colon cancer patients and analyzed its correlation with the clinical characteristics of colon cancer patients;miRBase was used to screen out the potential target gene of miR-1197 was Smad3,meanwhile,luciferase reporter gene assay,qrt-pcr and Western blot were used to verify the targeted regulatory effect of miR-1197 on Smad3;Western blot was used to detect the Smad3 expression in colon cancer tumor tissues and adjacent normal tissues;The effects of miR-1197 and Smad3 on the proliferation,cycle,invasion,migration of SW620 cells were detected by CCK8,flow cytometry,Tranwell invasion assay and wound healing assay,respectively.The tumor model of colon cancer xenograft in nude mice was established to detect the effect of miR-1197 on tumor growth and cell proliferation in vivo.Results The expression level of miR-1197 in serum was negatively correlated with the clinical indicators of colon cancer.The expression level of miR-1197 in the colon cancer tumor tissue was significantly lower than that in the adjacent normal tissue,and the expression level in the tumor tissue with distant metastasis was significantly lower than that in the non-metastatic tumor tissue.miR-1197 regulated the mRNA and protein expression of Smad3 by targeting the 3′-UTR of Smad3.The expression level of Smad3 in tumor tissues was significantly higher than that in adjacent normal tissues.miR-1197 inhibited the proliferation,invasion,migration and S phase cell aggregation of SW620 cells,but this inhibitory effect could be reversed by Smad3.miR-1197 inhibited the growth and proliferation of transplanted tumor cells in nude mice.Conclusion miR-1197 plays a tumor suppressive role in colon c

关 键 词：结肠癌 miR-1197 SMAD3 

分 类 号：R735.35[医药卫生—肿瘤]