miR-146a调控铁死亡过程对甲状腺乳头状癌细胞增殖和转移的调控作用
miR-146a regulates iron death process onproliferation and metastasis of papillary thyroid cancer cells
龚烨 1王胄 1苏鹏程1
作者信息
- 1. 新疆维吾尔自治区人民医院乳腺甲状腺科,乌鲁木齐 830001
- 折叠
摘要
目的 探讨微小核糖核酸-146a(miR-146a)通过影响铁死亡过程对人甲状腺癌(TPC-1)细胞增殖和转移的调控作用.方法 在培养箱内对TPC-1细胞孵育及传代,制备TPC-1单细胞悬液进行转染,转染后分为Control组(未处理)、miR-146a mimics NC组(进行非特异性对照miR-146a模拟物处理)、miR-146a mimics组(miR-146a模拟物处理)、inhibitor NC组(非特异性对照的miR-146a 抑制剂处理)、miR-146a inhibitor 组(miR-146a 抑制剂处理)、miR-146a inhibitor+Eras-tin(10 µmol/L)组(miR-146a抑制剂和Erastin处理)6组,采用细胞活力检测试剂(CCK8)、细胞迁移实验(Transwell)检测TPC-1细胞侵袭能力,划痕实验检测TPC-1细胞愈合能力,流式细胞术检测活性氧(ROS)表达情况,试剂盒检测TPC-1细胞中亚铁离子(Fe2+)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶4(GPX4)的表达变化.结果 miR-146a mimics组TPC-1细胞增殖活性最低,miR-146a inhibitor组TPC-1细胞增殖活性最高,差异有统计学意义(P＜0.05).miR-146a mimics组TPC-1细胞迁移距离和侵袭能力最低,miR-146a inhibitor组TPC-1细胞迁移距离和侵袭能力最高,差异有统计学意义(P＜0.05).miR-146a mimics组TPC-1细胞中Fe2+和ROS水平最高,GPX4和GSH水平最低,差异有统计学意义(P＜0.05).miR-146a inhibitor组TPC-1细胞中Fe2+和ROS水平最低,GPX4和GSH水平最高,差异有统计学意义(P＜0.05).与Control组比较,miR-146a inhibitor 组和 miR-146a inhibitor+Erastin 组 TPC-1 细胞增殖活性、迁移距离和侵袭能力均增大,Fe2+和ROS的表达降低,GSH和GPX4的表达升高,差异有统计学意义(P＜0.05).与 miR-146a inhibitor 组比较,miR-146a inhibitor+Erastin 组 TPC-1 细胞增殖活性、迁移距离和侵袭能力均减小,Fe2+和 ROS的表达升高,GSH和GPX4的表达降低,差异有统计学意义(P＜0.05).结论 miR-146a通过影响铁死亡的模式调控并影响TPC-1细胞增殖和转移的进程.
Abstract
Objective To investigate the regulatory effect of microrna-146a(miR-146a)on proliferation and metastasis of human thyroid cancer cell(TPC-1)by affecting the process of iron death.Methods TPC-1 cells were incubate and passedin the incubator,and TPC-1 single-cell suspension was preparedfor transfec-tion.Transfection was dividedinto control group(nottreated),miR-146a mimics NC group(treated with non-specific control miR-146a mimetics),miR-146a mimics group(treated with miR-146a mimetics)and inhibitor NC group(treated with non-specific control miR-146a inhibitors)and miR-146a inhibitor group(treated with miR-146a inhibitors),MiR-146a inhibitor+Erastin(10 μmol/L group)(treated with miR-146a inhibitor and Erastin)6 groups,cell viability assay(CCK8)and cell migration assay(Transwell)were used to detect the invasion ability of TPC-1 cells,scratch assay was used to detect the healing ability of TPC-1 cells,flow cytometry was used to detect the expression of reactive oxygen species(ROS).The expressionof ferrous ions(Fe2+),glutathione(GSH)and glutathione peroxidase 4(GPX4)in TPC-1 cells was detected by a kit.Results The miR-146a mimics group had the lowest proliferation activity of TPC-1 cells,while the miR-146a inhibitor group had the highest proliferation activity of TPC-1 cells,with a sta-tistically significant difference(P＜0.05).The miR-146a mimics group had the lowest migration distance and invasion ability of TPC-1 cells,while the miR-146a inhibitor group had the highest migration distance and invasion ability of TPC-1 cells,with statistically significant differences(P＜0.05).The miR-146a mimics group had the highest levels of Fe2+and ROS in TPC-1 cells,and the lowest levels of GPX4 and GSH,with statistically significant differences(P＜0.05).The miR-146a inhibitor group had the lowest levels of Fe2+and ROS in TPC-1 cells,and the highest levels of GPX4 and GSH,with statistically signifi-cant differences(P＜0.05).Compared with the control group,bothmiR-146a inhibitor group and miR-146a inhibitor+Erastin group showed an increase in TPC-1 cell proliferation activity,migration distance-and invasion ability.The expression of Fe2+and ROS was decreased,while the expression of GSH and GPX4 wasincreased,with statistical significance(P＜0.05).Compared with miR-146a inhibitor group,the miR-146a inhibitor+Erastin group showed a decrease in TPC-1 cell proliferation activity,migration distanceand invasion ability,an increase in Fe2+and ROS expression,and a decrease in GSH and GPX4 ex-pression,with statistical significance(P＜0.05).Conclusion miR-146a regulates and influences the pro-liferation and metastasis of TPC-1 cells by influencing the pattern of ferroptosis.
关键词
乳头状甲状腺癌/微小核糖核酸/铁死亡/铁死亡诱导剂/侵袭Key words
papillary thyroid cancer/microribonucleic acid/ferroptosis iron death inducers/invasion引用本文复制引用
基金项目
新疆维吾尔自治区自然科学基金资助项目(2022D01C140)
出版年
2023
NETL
NSTL
维普
万方数据