CHMP2B高表达促进胰腺癌发生和发展的机制  

High expression of CHMP2B promotes the occurrence and development of pancreatic cancer and explores its mechanism

作　　者：曹天宇 金皓东 黄灿 Cao Tianyu;Jin Haodong;Huang Can(Dept of Biological Chemistry and Basic Medical College,Anhui Medical University,Hefei 230032;The First Clinical Medical College,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学基础医学院生物化学与分子生物学教研室,合肥230032 [2]安徽医科大学第一临床医学院,合肥230032

出　　处：《安徽医科大学学报》2024年第9期1551-1556,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82203424)。

摘　　要：目的解析带电多泡体蛋白2B(CHMP2B)在胰腺癌中高表达的临床意义,探究CHMP2B在胰腺癌中的促癌机制。方法通过GEPIA网站的转录组学分析胰腺癌中CHMP2B的表达情况,探究CHMP2B的表达与胰腺癌无疾病生存率和总生存率的关系。通过Western blot法检测HPNE细胞与BXPC-3、MIA PaCa-2、PANC-1三种胰腺癌细胞中CHMP2B的表达。胰腺癌在MIA PaCa-2细胞中使用Lipofectamine 2000转染CHMP2B因子的过表达质粒,通过Western blot验证在MIA PaCa-2中过表达CHMP2B质粒的过表达效果。通过克隆形成实验比较过表达CHMP2B因子对MIA PaCa-2细胞成瘤能力。CCK-8实验和Transwell实验验证过表达CHMP2B因子对MIA PaCa-2细胞增殖和迁移侵袭的影响。采用免疫荧光染色检测CHMP2B因子的细胞定位,Western blot法检测CHMP2B高表达对微管关联蛋白1轻链3B(MAP1LC3B)和重组核孔蛋白62(P62)表达的影响。结果CHMP2B因子在胰腺癌组织中表达升高(P<0.01)。CHMP2B因子的过表达降低胰腺癌患者的总生存率和无疾病生存率(P<0.01)。人胰腺癌细胞系中CHMP2B表达量明显高于正常人胰腺导管上皮细胞。CHMP2B的高表达促进胰腺癌MIA PaCa-2细胞成瘤,同时与对照组相比CHMP2B高表达导致胰腺癌增殖、迁移和侵袭能力升高。免疫荧光染色可见CHMP2B在细胞核和细胞质中均有分布,并与膜泡分泌相关。CHMP2B的高表达促进了MAP1LC3B的表达,但是抑制了P62的表达。结论临床胰腺癌组织样本中CHMP2B的高表达与体外胰腺癌细胞系中的高表达相一致。CHMP2B分布在细胞核和细胞质,并且参与膜泡运输和溶酶体自噬,在胰腺癌细胞系MIA PaCa-2中,CHMP2B与自噬标志物MAP1LC3B表达呈正相关,并且抑制P62的表达。可知CHMP2B对胰腺癌恶性程度的促进与细胞自噬相关。CHMP2B通过调节膜泡运输和细胞自噬满足胰腺癌细胞高的营养代谢需求,进而促进胰腺癌的恶性生物学行为。Objective To analyze the clinical significance of elevated CHMP2B expression in pancreatic cancer,and to explore the oncogenic mechanism of CHMP2B in pancreatic cancer.Methods A transcriptomic analysis conducted on the GEPIA website revealed CHMP2B expression in over 20 types of malignant tumors.The relationship between CHMP2B expression and disease-free survival and overall survival of pancreatic cancer was analyzed.The correlation between CHMP2B,MAP1LC3B and P62 expression was analyzed by TCGA database.The overexpression plasmid of CHMP2B factor was transfected with Lipofectamine 2000 in MIA PaCa-2 cells,and the overexpression of CHMP2B factor in MIA PaCa-2 was verified by Western blot.The effect of overexpression of CHMP2B on tumorigenic ability of MIA PaCa-2 cells was verified by clonal formation experiment.The effect of overexpression of CHMP2B on proliferation,migration and invasion of MIA PaCa-2 cells was verified by CCK-8 and Transwell experiments.The localization of CHMP2B factor was detected by immunofluorescence staining.The correlation between CHMP2B,MAP1LC3B and P62 expression was analyzed by TCGA database.The expression of MAP1LC3B and P62 in CHMP2B overexpression was verified by qPCR.Results The expression of CHMP2B factor increased in pancreatic cancer(P<0.01).Overexpression of CHMP2B factor was associated with overall survival and disease-free survival(P<0.01).The high expression of CHMP2B promoted the tumorigenesis of MIA PaCa-2 cells in pancreatic cancer,and also promoted the proliferation,migration and invasion of pancreatic cancer.Immunofluorescence staining showed that CHMP2B was distributed in both nucleus and cytoplasm and correlated with vesicle secretion.Moreover,CHMP2B was positively correlated with the expression of autophagy marker MAP1LC3B while inhibiting the expression of P62.Conclusion The high expression of CHMP2B in clinical pancreatic cancer tissue samples is consistent with the high expression in vitro pancreatic cancer cell lines.CHMP2B is localized in both nucleus and cytopl

关 键 词：CHMP2B 膜泡运输 胰腺癌 溶酶体自噬 P62 MAP1LC3B 

分 类 号：R735.3[医药卫生—肿瘤]