摘要
目的探讨含SPOC结构域蛋白1(SPOC domain-containing protein 1,SPOCD1)在结直肠癌中的表达及相关分子机制。方法利用TCGA数据库和GEO数据库分析结直肠癌和癌旁正常组织中SPOCD1 mRNA的表达;采用免疫组化和Western blot法检测结直肠癌和癌旁正常组织或细胞系中SPOCD1蛋白表达,并分析其与临床病理特征的关系;应用GEPIA和Prognoscan数据库分析SPOCD1表达与结直肠癌患者预后的关系;基因集富集分析SPOCD1可能参与的通路;采用GEPIA数据库分析基因表达之间的相关性;利用siRNA沉默结直肠癌细胞系HCT116中SPOCD1蛋白表达,Western blot法验证下游基因。结果TCGA和GEO数据库分析结果显示,SPOCD1 mRNA在结直肠癌组织中的表达显著增加;免疫组化结果显示,SPOCD1蛋白在结直肠癌组织中高表达(67.8%,40/59),显著高于癌旁正常组织(37.3%,22/59)(P<0.001);SPOCD1在结直肠癌细胞系(HCT116和LoVo)中的表达高于正常结直肠细胞系(NCM460);SPOCD1蛋白表达与结直肠癌淋巴结转移、脉管浸润密切相关(P<0.05);生存分析显示,SPOCD1高表达组患者的总生存率和无瘤生存率均明显低于低表达组(P<0.05);基因集富集分析显示,SPOCD1可能参与上皮-间充质转化(epithelial-mesenchymal transition,EMT);SPOCD1与Slug表达呈正相关;Western blot结果示,与对照组相比,敲低组中SPOCD1和Slug蛋白水平均下降。结论SPOCD1在结直肠癌中高表达,并可能通过EMT通路促进结直肠癌的进展,有望作为结直肠癌新的诊断标志物及药物治疗新靶点。
Purpose To observe the expression and possible molecular mechanisms of SPOC domain-containing protein 1(SPOCD1)in colorectal cancer(CRC).Methods The expression of SPOCD1 mRNA in normal and CRC tissues was analyzed by using TCGA data and GEO data.Immunohistochemical staining and Western blot were used to detect the expression of SPOCD1 protein in CRC and normal tissues and cell lines.And the correlation between SPOCD1 gene expression and clinicopathological characters of CRC patients were analyzed.GEPIA and Prognoscan database were used to analyze relationship between the expression of SPOCD1 mRNA and the survival of CRC patients.GSEA was used to analyze the possible pathways involved by SPOCD1.The correlation of gene expression was analysed by using GEPIA database.Furthermore,small interfering RNA(siRNA)was used to knockdown SPOCD1 expression in HCT116.Western blot was used to validate the downstream gene.Results The analysis of TCGA and GEO data showed that the expression of SPOCD1 mRNA was significantly increased in CRC.The results of immunohistochemistry showed that the positive rate of SPOCD1 protein in CRC(67.8%,40/59)was significantly higher than that in normal tissues(37.3%,22/59)(P<0.001).The expression of SPOCD1 in colorectal cancer cell lines HCT116 and LoVo were higher than that in normal colorectal cell line NCM460.The expression of SPOCD1 showed significant correlation with lymph node metastasis and vascular infiltration(P<0.05).The overall survival rate and disease free survival rate was significantly lower in CRC patients with high SPOCD1 expression than those with low SPOCD1 expression(P<0.05).GSEA analysis showed that SPOCD1 may participate in the epithelial-mesenchymal transition(EMT)pathway.SPOCD1 expression is remarkably positively correlated with the expression of Slug.Western blot showed that SPOCD1 and Slug protein level decreased in the knockdown group compared with the control group.Conclusion SPOCD1 is highly expressed in CRC and may promote the progression of CRC via EMT pathway.It may be used as a new biomarker for diagnosis of CRC and a new target for drug therapy.
作者
温奕巽
李庆
田壮壮
白真真
陈静
曹立宇
WEN Yi-xun;LI Qing;TIAN Zhuang-zhuang;BAI Zhen-zhen;CHEN Jing;CAO Li-yu(Department of Pathology,Anhui Medical University,Hefei 230032,China;Department of Pathology,Fuyang Hospital of Anhui Medical University,Fuyang 236000,China)
出处
《临床与实验病理学杂志》
CAS
北大核心
2023年第7期802-806,共5页
Chinese Journal of Clinical and Experimental Pathology
基金
安徽省高校自然科学研究重点项目(KJ2021A0338)。
