SPTBN1对结直肠腺癌细胞及其多倍体肿瘤巨细胞生物学行为的影响及分子机制

Effects and molecular mechanisms of SPTBN1 on the biological behavior of colorectal cancer cells and polyploid tumor giant cells

目的探讨SPTBN1在结直肠腺癌细胞及其多倍体肿瘤巨细胞(polyploid giant cancer cells,PGCCs)中的作用及分子机制。方法应用CPTAC数据库分析结直肠腺癌组织和癌旁正常组织中SPTBN1蛋白表达水平;应用Kaplan-Meier Plotter、PrognoScan和UALCAN数据库分析SPTBN1蛋白表达与结直肠腺癌患者生存率的关系;应用Western blot法检测SPTBN1在结直肠腺癌组织和PGCCs中的表达量;应用平板克隆实验和划痕实验检测细胞的增殖与迁移能力;利用siRNA沉默PTPRN2和SPTBN1蛋白的表达。结果CPTAC数据库分析显示,SPTBN1蛋白在结直肠腺癌中的表达明显降低,其低表达与结直肠腺癌临床分期呈正相关;生存分析结果显示,SPTBN1低表达组患者的总生存率显著低于高表达组;应用奥沙利铂诱导的HCT116-PGCCs及LOVO-PGCCs,其增殖和迁移能力强于对照组HCT116和LOVO细胞;Western blot结果显示,SPTBN1蛋白在结直肠腺癌组织和PGCCs中的表达明显下降;SPTBN1敲低组HCT116-PGCCs及LOVO-PGCCs细胞的增殖和迁移能力增强;GEPIA数据库结果显示,SPTBN1蛋白表达与PTPRN2、E-cadherin蛋白表达呈正相关;敲低SPTBN1后,HCT116-PGCCs及LOVO-PGCCs细胞中E-cadherin蛋白表达水平下降;敲低PTPRN2后,HCT116-PGCCs及LOVO-PGCCs细胞中SPTBN1、E-cadherin蛋白表达水平明显降低。结论SPTBN1在结直肠腺癌和PGCCs中低表达,敲低SPTBN1可促进HCT116-PGCCs及LOVO-PGCCs细胞的增殖与迁移,其机制可能与PTPRN2/SPTBN1/E-cadherin通路相关。

Purpose To investigate the role and molecular mechanism of SPTBN1 in colorectal cancer(CRC)cells and its polypoid giant cancer cells(PGCCs).Methods CPTAC database was used to analyze the protein expression level of SPTBN1 in colorectal cancer and normal tissues.The relationship between SPTBN1 expression and prognosis in CRC patients was explored by Kaplan-Meier Plotter,PrognoScan and UALCAN databases.Western blot assay was conducted to detect the protein expression of SPTBN1 in CRC and PGCCs.The proliferation and migration of cells were analyzed using plate cloning and scratch experiment.The expression of PTPRN2 and SPTBN1 proteins were silenced by siRNA.Results CPTAC database analysis showed that SPTBN1 protein expression was significantly decreased in CRC,and its low expression was positively correlated with the clinical stage of CRC.Survival analysis results showed that the overall survival rate of CRC patients with low SPTBN1 expression group was significantly lower than that of patients with high SPTBN1 expression group.HCT116-PGCCs and LOVO-PGCCs were induced by oxaliplatin,and their proliferation and migration abilities were stronger than HCT116 and LOVO.SPTBN1 protein expression was distinctly decreased in colorectal cancer tissues and PGCCs by western blot assay.The proliferation and migration abilities of HCT116-PGCCs and LOVO-PGCCs were decreased in SPTBN1 knockdown group.GEPIA database showed that SPTBN1 protein was positively correlated with PTPRN2 and E-cadherin protein.In addition,the expression of E-cadherin in HCT116-PGCCs and LOVO-PGCCs was decreased after SPTBN1 knockdown.When PTPRN2 was knockdown by siRNA,the expression levels of SPTBN1 and E-cadherin in HCT116-PGCCs and LOVO-PGCCs were markedly declined.Conclusion SPTBN1 expression was low in colorectal cancer and PGCCs.The group of SPTBN1-knockdown can promote the proliferation and migration of HCT116-PGCCs and LOVO-PGCCs,and the mechanism may be related to PTPRN2/SPTBN1/E-cadherin pathway.