摘要
目的探讨食管鳞癌(esophageal squamous cell carcinoma,ESCC)中CBX4的表达情况,揭示CBX4对ESCC细胞增殖的影响及其潜在机制。方法TCGA数据库中分析CBX4在不同肿瘤中的表达情况;GEO在线数据库对ESCC及其对应癌旁组织CBX4的表达水平进行t检验分析;过表达或敲低CBX4后,通过MTT、菌落集成实验和流式凋亡术检测CBX4对ESCC细胞活力的影响;裸鼠皮下成瘤和免疫组化法验证CBX4对瘤体增殖的影响;qRT-PCR和Western blot方法验证凋亡相关基因PARP、Bcl-2、Bax的mRNA和蛋白表达水平;筛选转录组测序结果推测CBX4可能的作用机制,并通过qRT-PCR和Western blot进行验证。结果CBX4在多种肿瘤中高表达;ESCC组织中CBX4表达高于正常组织。敲低CBX4后ESCC细胞凋亡增加,增殖被抑制(P<0.05);同时,被剪切的PARP和Bax抑癌基因mRNA和蛋白表达水平升高,促癌基因Bcl-2表达水平降低。体内裸鼠成瘤结果表明,过表达CBX4后,瘤体体积和Ki67表达明显增加(P<0.05)。转录组测序结果表明CBX4与p38 MAPK通路基因相关性高,敲低CBX4后,p38 MAPK通路被激活,其下游转录因子表达量增加(P<0.05),从而诱导ESCC细胞凋亡。结论CBX4在ESCC中高表达,发挥促癌作用,可能通过调控p38 MAPK信号通路影响ESCC细胞增殖。
Aim To investigate the expression level of CBX4 in esophageal squamous cell carcinoma(ESCC)and the effect of CBX4 on ESCC proliferation and underlying molecular mechanisms.Methods The expression of CBX4 in different cancers was analyzed in Pan-cancers.The expression level of CBX4 in ESCC was analyzed by t-test based on Gene Expression Omnibus(GEO)data.The viability of CBX4-overexpressed/knockdown ESCC cells was detected by MTT assay,colony formation assay and flow cytometry assay.Furthermore,the tumor volumn,tumor weight and Ki67 expression were measured by mouse xenograft assay and immunohistochemistry.The mRNA and protein expression levels of apoptosis-related genes PARP、Bcl-2、Bax were determined by qRT-PCR and Western blot,respectively.In addition,the underlying molecular mechanism of CBX4 in ESCC was revealed by qRT-PCR and Western blot.Results CBX4 was upregulated in various cancers.The expression level of CBX4 in ESCC was higher than that in normal tissues(P<0.05)based on Gene Expression Omnibus(GEO)data.Compared with the normal group,the proliferation of CBX4 knockdown ESCC cells was significantly inhibited and the apoptosis was promoted(P<0.05).Meanwhile,the mRNA and protein expression levels of cleaved PARP and Bax were upregulated while that of Bcl-2 was downregulated.In CBX4 overexpression group,tumor volume in vivo increased(P<0.05).Immunohistochemical results also showed an increase in Ki67 expression.Furthermore,the results of RNA-seq,bioinformatics analysis and qRT-PCR experiments indicated that CBX4 probably regulated the proliferation and apoptosis of ESCC through p38 MAPK signaling pathway.Conclusion CBX4 is highly expressed in ESCC and plays as an oncogene role,which might regulate cell proliferation through the p38 MAPK signaling pathway.
作者
马燕春
花雨艳
刘蕊
毋阿婧
尹晓洁
杨洁
MA Yan-chun;HUA Yu-yan;LIU Rui;WU A-jing;YIN Xiao-jie;YANG Jie(Translational Medicine Research Center,Shanxi Medical University,Taiyuan 030001,China;Second Hospital of Shanxi Medical University,Shanxi Medical University,Taiyuan 030001,China;College of Pharmacy,Shanxi Medical University,Taiyuan 030001,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2024年第9期1673-1679,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81802440)
山西省基础研究计划自然科学研究面上项目(No 2022030212211187)。
关键词
食管鳞癌
CBX4
增殖
凋亡
p38
MAPK信号通路
药物靶点
esophageal squamous cell carcinoma
CBX4
proliferation
apoptosis
p38 MAPK signaling pathway
drug target