目的:筛选三阴性乳腺癌(triple-negative breast cancer,TNBC)潜在生物标志物并进行验证,探讨TNBC预后差、难治疗的遗传因素。方法:GEO与TCGA下载TNBC表达谱数据,使用GEO2R与R语言筛选差异表达基因(differentially expressed genes,DEGs...目的:筛选三阴性乳腺癌(triple-negative breast cancer,TNBC)潜在生物标志物并进行验证,探讨TNBC预后差、难治疗的遗传因素。方法:GEO与TCGA下载TNBC表达谱数据,使用GEO2R与R语言筛选差异表达基因(differentially expressed genes,DEGs);KEGG与GO富集分析;STRING数据库与Cytoscape软件绘制PPI网络并筛选核心基因;Kaplan-Meier数据库绘制生存曲线;利用bc-GenExMiner与UALCAN数据库验证以上结果;CIBERSORT算法与TIMER数据库分析预后相关基因对免疫细胞浸润及免疫检查点基因表达量的影响;qRT-PCR与Western Blot鉴定目的基因在TNBC与非三阴性乳腺癌(non-triple-negative breast cancer,n-TNBC)间的表达差异;si-RNA敲减目的基因;划痕实验与Transwell实验对比目的基因敲减与否对癌细胞侵袭、迁移的影响。结果:共得到123个TNBC相关DEGs,主要与雌激素信号通路相关;MCODE聚类分析筛选出16个核心基因;KRT6B、CDH3与预后相关,其中KRT6B在TNBC中高表达;KRT6B能够减少γδT细胞浸润,使γδT细胞表面CD27与PTPRC(CD45)减少;KRT6B表达量与CD276(B7-H3)表达量呈正相关;qRT-PCR与Western Blot结果显示KRT6B在TNBC中高表达;KRT6B经敲减后TNBC细胞侵袭、迁移能力均明显减弱。结论:KRT6B在TNBC中高表达,可能通过抑制γδT细胞浸润、减少γδT细胞表面标志物的表达并下调免疫检查点CD276的表达,与TNBC不良预后相关,有望成为诊断与治疗的新靶点。展开更多
Objectives:Keratin 6A(KRT6A)has been implicated in the progression of multiple malignancies;however,its expression pattern and biological role in cervical cancer(CC)have not been elucidated.This study aims to investig...Objectives:Keratin 6A(KRT6A)has been implicated in the progression of multiple malignancies;however,its expression pattern and biological role in cervical cancer(CC)have not been elucidated.This study aims to investigate KRT6A expression in CC tissues and evaluate its effects on cellular proliferation,migration,and invasion,thereby assessing its potential as a biomarker and therapeutic target.Methods:Differentially expressed genes were screened from the Gene Expression Omnibus(GEO)dataset(GSE9750)using the thresholds∣log2FC∣>2 and false discovery rate(FDR)<0.05.Immunohistochemistry was performed to evaluate KRT6A protein expression in tumor tissues.Stable KRT6A knockdown was established in CC cell lines to assess proliferation,colony formation,migration,and invasion in vitro.A nude-mouse xenograft model was used to evaluate tumor growth in vivo.Results:KRT6A expression was significantly increased in CC tissues relative to adjacent non-tumor tissues(p=0.019).Patients with KRT6A-positive tumors had a significantly lower 3-year progression-free survival(PFS)rate than those with KRT6Anegative tumors(45.16%vs.78.57%;p=0.009).KRT6A was identified as an independent prognostic indicator for CC(p=0.044).In vitro,KRT6A silencing significantly reduced colony formation,proliferation,migration,and invasion in SiHa and HeLa cells in comparison to controls(p<0.05).In vivo,tumors in the KRT6A knockdown group were significantly smaller,with reduced expression of both KRT6A and Ki-67 in tumor tissues(p<0.05).Conclusion:KRT6A is overexpressed in CC and associates with adverse clinicopathological features and poor PFS.Knockdown of KRT6A suppresses cell proliferation,migration,and invasion,indicating that KRT6A may represent a promising prognostic biomarker and potential target for treating CC.展开更多
文摘目的:筛选三阴性乳腺癌(triple-negative breast cancer,TNBC)潜在生物标志物并进行验证,探讨TNBC预后差、难治疗的遗传因素。方法:GEO与TCGA下载TNBC表达谱数据,使用GEO2R与R语言筛选差异表达基因(differentially expressed genes,DEGs);KEGG与GO富集分析;STRING数据库与Cytoscape软件绘制PPI网络并筛选核心基因;Kaplan-Meier数据库绘制生存曲线;利用bc-GenExMiner与UALCAN数据库验证以上结果;CIBERSORT算法与TIMER数据库分析预后相关基因对免疫细胞浸润及免疫检查点基因表达量的影响;qRT-PCR与Western Blot鉴定目的基因在TNBC与非三阴性乳腺癌(non-triple-negative breast cancer,n-TNBC)间的表达差异;si-RNA敲减目的基因;划痕实验与Transwell实验对比目的基因敲减与否对癌细胞侵袭、迁移的影响。结果:共得到123个TNBC相关DEGs,主要与雌激素信号通路相关;MCODE聚类分析筛选出16个核心基因;KRT6B、CDH3与预后相关,其中KRT6B在TNBC中高表达;KRT6B能够减少γδT细胞浸润,使γδT细胞表面CD27与PTPRC(CD45)减少;KRT6B表达量与CD276(B7-H3)表达量呈正相关;qRT-PCR与Western Blot结果显示KRT6B在TNBC中高表达;KRT6B经敲减后TNBC细胞侵袭、迁移能力均明显减弱。结论:KRT6B在TNBC中高表达,可能通过抑制γδT细胞浸润、减少γδT细胞表面标志物的表达并下调免疫检查点CD276的表达,与TNBC不良预后相关,有望成为诊断与治疗的新靶点。
基金supported by Yixing Taodu Light Science and Technology Research Plan,Grant Number:2024SF14.
文摘Objectives:Keratin 6A(KRT6A)has been implicated in the progression of multiple malignancies;however,its expression pattern and biological role in cervical cancer(CC)have not been elucidated.This study aims to investigate KRT6A expression in CC tissues and evaluate its effects on cellular proliferation,migration,and invasion,thereby assessing its potential as a biomarker and therapeutic target.Methods:Differentially expressed genes were screened from the Gene Expression Omnibus(GEO)dataset(GSE9750)using the thresholds∣log2FC∣>2 and false discovery rate(FDR)<0.05.Immunohistochemistry was performed to evaluate KRT6A protein expression in tumor tissues.Stable KRT6A knockdown was established in CC cell lines to assess proliferation,colony formation,migration,and invasion in vitro.A nude-mouse xenograft model was used to evaluate tumor growth in vivo.Results:KRT6A expression was significantly increased in CC tissues relative to adjacent non-tumor tissues(p=0.019).Patients with KRT6A-positive tumors had a significantly lower 3-year progression-free survival(PFS)rate than those with KRT6Anegative tumors(45.16%vs.78.57%;p=0.009).KRT6A was identified as an independent prognostic indicator for CC(p=0.044).In vitro,KRT6A silencing significantly reduced colony formation,proliferation,migration,and invasion in SiHa and HeLa cells in comparison to controls(p<0.05).In vivo,tumors in the KRT6A knockdown group were significantly smaller,with reduced expression of both KRT6A and Ki-67 in tumor tissues(p<0.05).Conclusion:KRT6A is overexpressed in CC and associates with adverse clinicopathological features and poor PFS.Knockdown of KRT6A suppresses cell proliferation,migration,and invasion,indicating that KRT6A may represent a promising prognostic biomarker and potential target for treating CC.
