miR-146a-5p、SMAD4在甲状腺滤泡癌组织中表达变化和对FTC-238细胞系增殖迁移侵袭影响及靶向关系OACSTPCD
Expression changes of miR-146a-5p and SMAD4 in thyroid follicular carcinoma tissues,their effects on proliferation,migration,and invasion of FTC-238 cells,and verification of the targeted relationship
目的 通过观察甲状腺滤泡癌(FTC)组织中miR-146a-5p、SMAD4的表达变化及对细胞增殖、迁移、侵袭能力的影响,分析miR-146a-5p与SMAD4之间的靶向关系,以探讨miR-146a-5p是否通过靶向SMAD4促进FTC增殖、迁移、侵袭.方法 收集FTC组织标本30例份,非肿瘤甲状腺或距离肿瘤边缘>2 cm 癌旁组织30例份,采用RT-qPCR法检测FTC组织及癌旁组织中miR-146a-5p、SMAD4 mRNA.将FTC-238细胞系分为miR-146a-5p mimics组和阴性对照(miR-NC组)分别转染miR-146a-5p mimics(使细胞过表达miR-146a-5p)及miR-NC.采用CCK8法观察两组培养12、24、48 h时的细胞增殖能力,划痕实验观察两组细胞迁移能力,Transwell小室实验观察两组细胞侵袭能力.分别采用RT-qPCR及免疫印迹法检测两组细胞中SMAD4 mRNA及蛋白.双荧光素酶报告基因实验验证miR-146a-5p与SMAD4的靶向关系.结果 与癌旁组织相比,FTC组织中miR-146a-5p相对表达量降低(P<0.05);SMAD4 mRNA相对表达量升高(P<0.05).与miR-NC组相比,各个时间点miR-146a-5p mimics组细胞OD值降低(P 均<0.05);与miR-NC组相比,miR-146a-5p mimics组细胞迁移、侵袭数目减少(P 均<0.05).与miR-NC组相比,miR-146a-5p mimics组SMAD4 mRNA及蛋白相对表达量降低(P均<0.05).miR-146a-5p与SMAD4存在靶向关系.结论 FTC组织中miR-146a-5p表达降低、SMAD4表达升高;miR-146a-5p过表达可抑制FTC-238细胞系增殖、迁移、侵袭;miR-146a-5p与SMAD4之间存在靶向关系.miR-146a-5p可能通过靶向抑制SMAD4促进FTC癌细胞的增殖、迁移、侵袭.
Objective To analyze the targeted relationship between miR-146a-5p and SMAD4 by observing the ex-pression changes of microRNA-146a-5p(miR-146a-5p)and SMAD4 homolog 4(SMAD4)in follicular thyroid carcinoma(FTC)tissues and their effects on cell proliferation migration and invasion abilities,in order to explore whether miR-146a-5p promotes invasion and metastasis of FTC by targeting SMAD4.Methods We collected 30 samples of FTC tissues and 30 samples of non-tumor thyroid tissues or tissues adjacent to the tumor margin(>2 cm)from patients with FTC.We used RT-qPCR to detect miR-146a-5p and SMAD4 mRNA in the FTC tissues and adjacent tissues.The FTC-238 cells were di-vided into the miR-146a-5p mimics group and the negative control(miR-NC group),which were transfected with miR-146a-5p mimics(to overexpress miR-146a-5p in the cells)and miR-NC,respectively.CCK-8 was used to observe the cell proliferation abilities of the two groups at 12,24,and 48 h of culture.The scratch test was used to observe the migration abilities of the two groups,and the Transwell chamber experiment was used to observe the invasion abilities of the two groups.RT-qPCR and Western blotting were used to detect SMAD4 mRNA and protein in the two groups,respectively.The dual luciferase reporter gene experiment was used to verify the targeted relationship between miR-146a-5p and SMAD4.Results The relative expression level of miR-146a-5p in FTC tissues was lower than that in the adjacent tissues(P<0.05).The relative expression level of SMAD4 mRNA in FTC tissues was higher than that in adjacent tissues(P<0.05).The OD value at each time point in the miR-146a-5p mimics group was lower than that in the miR-NC group(all P<0.05).The number of migration and invasive cells after transfection in the miR-NC group was larger than that in the miR-146a mimics group(P<0.05).The relative expression levels of SMAD4 mRNA and protein in the miR-146a-5p mimics group were lower than those in the miR-NC group(both P<0.05).There was a targeted relationship between miR-146a-5p and SMAD4.Conclusion In FTC tissues,the expression of miR-146a-5p decreases while the expression of SMAD4 in-creases.Overexpression of miR-146a-5p can inhibit the proliferation,migration,and invasion of the FTC-238 cells.There is a targeted relationship between miR-146a-5p and SMAD4.MiR-146a-5p may promote the proliferation,invasion,and migration of FTC cells by targetedly inhibiting SMAD4.
黄睿;刘红春;王昌敏
新疆维吾尔自治区人民医院临床检验中心,乌鲁木齐 830001
临床医学
微小RNA-146a-5pSMAD同源物4细胞增殖细胞侵袭细胞转移甲状腺滤泡癌
microRNA-146a-5pSMAD homolog 4cell proliferationcell invasioncell metastasisthyroid fol-licular carcinoma
《山东医药》 2024 (027)
1-5 / 5
新疆维吾尔自治区自然科学基金项目(2023D01C59).
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