临床与病理杂志2025，Vol.45Issue(4)：405-415,11.DOI:10.11817/j.issn.2095-6959.2025.240605
UCHL1在结直肠癌细胞增殖和转移中的作用
Role of UCHL1 in the proliferation and metastasis of colorectal cancer cells
摘要
Abstract
Objective:Colorectal cancer(CRC)is the second leading cause of cancer-related deaths worldwide,with high metastatic potential and therapeutic resistance remaining major clinical challenges.In recent years,targeted modulation of key tumor molecules has emerged as a research focus,particularly the role of the ubiquitin-proteasome system(UPS)in tumorigenesis and progression.This study aims to investigate the mechanistic role of ubiquitin carboxyl-terminal hydrolase L1(UCHL1)in CRC progression. Methods:Tumor tissue specimens were collected from 74 CRC patients(June 2016 to June 2018)at the First Affiliated Hospital of Henan University of Chinese Medicine and Huai'an First People's Hospital,all untreated with radiotherapy or chemotherapy.The expression level of UCHL1 protein was analyzed by immunohistochemistry(IHC),and its positive classification results were records.Loss-of-function experiments were conducted in CRC cell lines(HCT 116 and LoVo),divided into a si-UCHL1(transfected with UCHL1-specific siRNA)group and a normal(negative control)group.Cell proliferation was evaluated using colony formation assays and thiazolyl blue tetrazolium bromide(MTT)assays,while cell migration and invasion capacities were assessed by quantifying transmembrane cells in Transwell assays.Expression changes of apoptosis-related proteins[B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),caspase-3,caspase-9,cytochrome c(cyt c)]and autophagy-related proteins[mechanistic target of rapamycin complex 2(mTORC2),phosphorylated protein kinase B(pAKT),forkhead box O3(FOXO3),microtubule-associated protein light chain 3B-Ⅱ(LC3BⅡ)]were analyzed by Western blotting. Results:Immunohistochemical(IHC)analysis revealed significantly elevated UCHL1 protein expression in both primary CRC tissues(89.10％,66/74)and metastatic CRC lesions(90.00％,18/20).Notably,high UCHL1 expression levels showed significant correlation with advanced tumor stage and poorer clinical outcomes in CRC patients.The colony formation assay demonstrated that UCHL1 knockdown significantly inhibited the proliferative capacity of CRC cells(P<0.05).MTT assay results demonstrated that UCHL1 knockdown significantly inhibited cellular viability in CRC cells(P<0.05).Transwell assay results demonstrated that compared to the normal group,the Si-UCHL1 group exhibited significant reductions in migratory and invasive cell numbers.Specifically,in HCT 116 cells,migration decreased from 530.23±7.07 to 275.30±5.42;invasion declined from 140.15±5.66 to 40.21±3.54;in LoVo cells,migration decreased from 560.11±8.48 to 189.12±6.33;invasion dropped from 280.10±6.20 to 36.25±3.46.Western blotting analysis revealed that compared with the Normal group,the Si-UCHL1 group showed significantly decreased protein expression levels of mTORC2,p-AKT,and Bcl-2(all P<0.05),while demonstrating markedly increased expression of FoxO3,LC3B-Ⅱ,Bax,cleaved caspase-3,cytochrome c,and cleaved caspase-9(all P<0.05),accompanied by distinct apoptotic features. Conclusion:UCHL1 promotes CRC progression by inhibiting apoptosis and autophagy,and it may serve as a molecular marker for CRC diagnosis and prognosis.Targeting UCHL1 could be a novel therapeutic strategy for CRC treatment.关键词
泛素羧基末端水解酶-1/自噬/凋亡/结直肠癌/转移/增殖Key words
UCHL1/autophagy/apoptosis/colorectal cancer/metastasis/proliferation引用本文复制引用
杨凯丽,周武碧,牛德亮,郭地,成树贞,刘磊,周艳阳,任伟宏..UCHL1在结直肠癌细胞增殖和转移中的作用[J].临床与病理杂志,2025,45(4):405-415,11.基金项目
河南省高等学校重点科研项目计划基础研究专项(19zx009).This work was supported by the Basic Research Special Project of Henan Provincial Key Scientific Research Projects Plan in Higher Education Institutions,China(19zx009). （19zx009）
