摘要
目的 探讨肌盲样蛋白2(muscleblind-like 2,MBNL2)通过p38 MAPK/mTOR信号通路调控食管鳞状细胞癌(esophageal squamous cell carcinoma, ESCC)细胞凋亡的分子机制。方法 利用GEPIA2肿瘤在线数据库分析MBNL2在ESCC癌组织中的差异表达,通过qRT-PCR验证MBNL2在ESCC细胞系KYSE-150、KYSE270、KYSE70中的mRNA表达水平,运用基因沉默/过表达技术结合克隆形成实验、Transwell迁移/侵袭实验评估MBNL2对细胞恶性表型的影响,Western blot检测MAPK/mTOR通路关键蛋白(p-MAPK、p-mTOR)及自噬-凋亡相关标志物(Bcl-2/Bax、LC3-II/I、p62)的表达变化,采用特异性MAPK抑制剂SB202190分析MAPK/mTOR通路的作用。结果 MBNL2在ESCC组织和细胞中高表达。沉默MBNL2抑制ESCC细胞增殖、迁移和侵袭,而过表达MBNL2则促进细胞增殖、迁移和侵袭数。同时,沉默MBNL2能下调p-p38 MAPK、p-mTOR、Bcl-2和p62蛋白表达水平,上调Bax蛋白水平和LC3II/LC3I比值,过表达MBNL2时则呈现相反趋势。p38 MAPK抑制剂SB202190能够逆转过表达MBNL2对细胞增殖、迁移、侵袭、凋亡和自噬的影响。结论 MBNL2通过激活p38 MAPK/mTOR通路阻断自噬,抑制ESCC细胞凋亡。本研究为开发新型ESCC治疗策略提供了新的思路和理论依据。
Objective To investigate the molecular mechanisms by which muscleblind-like 2(MBNL2)regulates apoptosis in esophageal squamous cell carcinoma(ESCC)cells through the p38 MAPK/mTOR signaling pathway.Methods The differential expression of MBNL2 in ESCC cancer tissues was analyzed using GEPIA2 online tumor database,and the mRNA expression levels of MBNL2 in normal esophageal squamous epithelial cells ET-1A and human ESCC cell lines KYSE-150,KYSE270 and KYSE70 were verified by qRT-PCR.Gene silencing and overexpression techniques were combined with clonogenic assays and Transwell migration/invasion assays to assess the effects of MBNL2 on the malignant phenotypes of the cells.Western blotting was used to detect the expression changes of key proteins in the p38 MAPK/mTOR pathway(p-p38 MAPK,p-mTOR)and autophagy-apoptosis related markers(Bcl-2/Bax,LC3-II/I,p62).The role of the p38 MAPK/mTOR pathway was analyzed using the specific MAPK inhibitor SB202190.Results MBNL2 was highly expressed in ESCC tissues and cells.Silencing MBNL2 inhibited cell proliferation,migration,and invasion of ESCC cells,while overexpression of MBNL2 promoted these processes.Silencing MBNL2 downregulated the levels of p-p38 MAPK,p-mTOR,Bcl-2 and p62,and upregulated the level of Bax and the ratio of LC3II/LC3I,with the opposite effects observed upon MBNL2 overexpression.The p38 MAPK inhibitor SB202190 can reverse the effects of overexpression of MBNL2 on cell proliferation,migration,invasion,apoptosis and autophagy.Conclusion MBNL2 blocks autophagy and inhibits apoptosis in ESCC cells by activating the p38 MAPK/mTOR pathway.This study provides new insights and theoretical basis for the development of novel therapeutic strategies for ESCC.
作者
戴晓燕
张燕
仇建伟
张圣来
Dai Xiaoyan;Zhang Yan;Qiu Jianwei;Zhang Shenglai(Department of Gastroenterology,The Second Affiliated Hospital of Nantong University,Nantong 226001,China)
出处
《中国组织化学与细胞化学杂志》
2025年第4期319-326,共8页
Chinese Journal of Histochemistry and Cytochemistry
基金
南通大学临床医学专项科研基金一般项目(2023LY002)。
