miR-449a下调TGIF2抑制甲状腺癌细胞CAL-62恶性增殖的研究  

Study of miR-449a down-regulating TGIF2 and inhibiting the malignant proliferation of thyroid cancer cell CAL-62

作　　者：岳秀杰 杨崔航 单妍 赵月 樊静 YUE Xiujie;YANG Cuihang;SHAN Yan;ZHAO Yue;FAN Jing(Department of Pathology,Qinhuangdao Fourth Hospital,Qinhuangdao 066000,Hebei,China)

机构地区：[1]秦皇岛市第四医院北区病理科,河北秦皇岛066000

出　　处：《西部医学》2024年第2期191-197,共7页Medical Journal of West China

基　　金：秦皇市科学技术研究与发展计项目(201902A076)。

摘　　要：目的 探讨miR-449a靶向TGIF2对甲状腺癌细胞CAL-62恶性增殖、氧化应激和上皮间质转化(EMT)的调控作用。方法 以甲状腺癌细胞CAL-62作为研究对象。分为对照组、miR-449a mimic组、pcDNA-TGIF2组、miR-449a mimic+pcDNA-TGIF2共转染组。采用RT-qPCR检测细胞中miR-449a和TGIF2表达;TargetScan软件与双荧光素酶报告实验验证二者靶向关系;CCK8检测细胞活力;Brdu染色检测细胞增殖;试剂盒检测氧化应激标记物SOD和MDA含量;DCF染色检测ROS含量;Western blot检测EMT标记分子E-cadherin, N-cadherin和Fibronectin的蛋白表达。结果 TGIF2在CAL-62细胞中表达较高,与对照组相比,pcDNA-TGIF2转染组BrdU阳性细胞数目和百分率显著增加,SOD含量显著升高而MDA和ROS含量显著降低,同时N-cadherin和Fibronectin表达显著上升而E-cadherin表达显著下降(均P<0.05)。在miR-449a mimic+pcDNA-TGIF2共转染组中上述现象被有效缓解和逆转。结论 miR-449a过表达可通过靶向TGIF2抑制甲状腺癌细胞CAL-62恶性增殖,加剧其氧化应激和上皮间质转换。Objective To investigate the effect of miR-449a targeting TGIF2(TGF-β-induced factor homeobox 2)on the malignant proliferation,oxidative stress and epithelial mesenchymal transition(EMT)of thyroid cancer cell line CAL-62.Methods CAL-62 was used as the research object.Group:control group,miR-449a mimic group,pcDNA-TGIF2 group and miR-449a mimic+pcDNA-TGIF2 co-transfection group.RT-qPCR was used to detect the expression of miR-449a and TGIF2.Targetscan software and double luciferase reporter assay were used to verify the targeted relationship;CCK8 was used to detect cell viability.BrdU staining was used to detect cell proliferation;kits were used to detect the content of oxidative stress markers SOD and MDA;DCF staining was used to detect ROS content;Western blot was used to detect the expression of EMT markers E-cadherin,N-cadherin and Fibronectin.Results TGIF2 was highly expressed in CAL-62 cells.Compared with the control group,the number and percentage of BrdU positive cells in pcDNA-TGIF2 transfection group were significantly increased,SOD content was significantly increased,MDA and ROS contents were significantly decreased,and the expression of N-cadherin and fibronectin was significantly increased,and the expression of E-cadherin was significantly decreased.In miR-449a mimic+pcDNA-TGIF2 co-transfection group,the above phenomenon was effectively alleviated and reversed.Conclusion Overexpression of miR-449a can inhibit the malignant proliferation of CAL-62 cells by targeting TGIF2,and aggravate oxidative stress and EMT.

关 键 词：甲状腺癌 miR-449a TGIF2 CAL-62细胞 DCF染色 上皮间质转化 

分 类 号：R736.1[医药卫生—肿瘤]