| 王潇玉,田阳子,吴振杰,郭 森,王思佳,郭伟楠.MiR-210在黑素瘤细胞铁死亡中的作用和机制研究[J].,2021,(10):1818-1823 |
| MiR-210在黑素瘤细胞铁死亡中的作用和机制研究 |
| The Investigation on the Role of MiR-210 in Ferroptosis and the Underlying Mechanism in Melanoma |
| 投稿时间：2020-10-23 修订日期：2020-11-18 |
| DOI：10.13241/j.cnki.pmb.2021.10.004 |
| 中文关键词: miR-210 TFRC 黑素瘤 铁死亡 |
| 英文关键词: miR-210 TFRC Melanoma Ferroptosis |
| 基金项目:国家自然科学基金项目（81902791 ; 81625020） |
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| 中文摘要: |
| 摘要 目的：探讨miR-210在黑素瘤细胞铁死亡中的作用及其可能调控机制。方法：通过qRT-PCR实验检测0.5 μM铁死亡诱导剂RSL3刺激前后黑素瘤细胞系A2058和451Lu中miR-210的表达水平变化情况；在0.5 μM铁死亡诱导剂RSL3刺激的同时，给予antagomiR-210处理抑制黑素瘤细胞系A2058和451Lu中miR-210的表达后，通过CCK8法检测细胞存活水平，通过流式细胞术检测细胞内lipid ROS水平；在此基础上，通过生物信息学分析、qRT-PCR实验、免疫印迹实验和萤光素酶报告实验明确miR-210对靶分子TFRC的调控作用；在0.5 μM铁死亡诱导剂RSL3刺激的条件下，给予antagomiR-210处理抑制miR-210的表达，同时使用siRNA沉默TFRC的表达水平，通过CCK8法检测细胞存活水平。结果：1）在0.5 μM RSL3刺激后，黑素瘤细胞系A2058和451Lu中miR-210的表达水平显著升高；2） 在0.5 μM RSL3刺激后，抑制miR-210的表达可加剧黑素瘤细胞系A2058和451Lu的死亡，同时显著增加细胞内lipid ROS水平；3）miR-210可直接结合TFRC mRNA的3'UTR区域，过表达miR-210可以显著抑制TFRC的转录和蛋白表达；4）在0.5 μM RSL3刺激的条件下，沉默TFRC的表达可以显著逆转抑制miR-210对黑素瘤细胞系A2058和451Lu铁死亡的促进作用。结论：miR-210是黑素瘤细胞铁死亡的重要抑制因子，且是通过靶向调控TFRC实现的。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the role and possible underlying mechanism of miR-210 in the regulation of ferroptosis in melanoma. Methods: qRT-PCR assay was used to detect the level of miR-210 in A2058 and 451Lu melanoma cell lines before and after the stimulation with 0.5 μM ferroptosis inducer RSL3. After the stimulation with 0.5 μM ferroptosis inducer RSL3, antagomiR-210 was used to obtain the knockdown of miR-210 level in A2058 and 451Lu melanoma cell lines, and cell survival was detected by CCK8 assay, as well as lipid ROS detected by flow cytometry analysis. On this basis, bioinformatics analysis, qRT-PCR assay, immunoblotting assay and luciferase report assay were used to clarify the regulatory mechanism of TFRC by miR-210. After the stimulation with 0.5 μM ferroptosis inducer RSL3, siRNA transfection was used to obtain the knockdown of TFRC expression in A2058 and 451Lu melanoma cells with the pre-treatment with antagomiR-210 that could suppress miR-210 level, and cell survival was then detected by CCK8 assay. Results: 1) MiR-210 was increased significantly in both A2058 and 451Lu melanoma cell lines after the stimulation with 0.5 μM ferroptosis inducer RSL3. 2) Under the stimulation with 0.5 μM ferroptosis inducer RSL3, the knockdown of miR-210 could significantly attenuate cell survival of A2058 and 451Lu melanoma cell lines, with the level of intracellular lipid ROS profoundly increased. 3) MiR-210 could directly bind to the 3'UTR region of TFRC mRNA. The overexpression of miR-210 could significantly suppress the expression of TFRC at both mRNA and protein levels in both A2058 and 451Lu melanoma cell lines. 4) The knockdown of TFRC could reverse the effect of silencing miR-210 on ferroptosis in both A2058 and 451Lu melanoma cell lines. Conclusion: MiR-210 is a crucial suppressor of ferroptosis in melanoma. MiR-210 inhibited melanoma cell ferroptosis via the suppression of its novel target TFRC. |
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