Pancreatic cancer has a dismal prognosis due to late detection and lack of efficient therapies.The Kirsten rat sarcoma virus(KRAS)oncogene is mutated in up to 90%of all pancreatic ductal adenocarcinomas(PDACs)and cons...Pancreatic cancer has a dismal prognosis due to late detection and lack of efficient therapies.The Kirsten rat sarcoma virus(KRAS)oncogene is mutated in up to 90%of all pancreatic ductal adenocarcinomas(PDACs)and constitutes an attractive target for therapy.However,the most common KRAS mutations in PDAC are G12D(44%),G12V(34%)and G12R(20%)that are not amenable to treatment by KRAS G12C-directed cysteine-reactive KRAS inhibitors such as Sotorasib and Adagrasib that exhibit clinical efficacy in lung cancer.KRAS G12C mutant pancreatic cancer has been treated with Sotorasib but this mutation is detected only in 2%–3%of PDAC.Recently,the KRAS G12D-directed MRTX1133 inhibitor has entered clinical trials and more of such inhibitors are in development.The other KRAS mutations may be targeted indirectly via inhibition of the cognate guanosine exchange factor(GEF)Son of Sevenless 1 that drives KRAS.These agents seem to provide the means to target the most frequent KRAS mutations in PDAC and to improve patient outcomes.展开更多
Background:Pancreatic ductal adenocarcinoma(PDAC)has a rich and complex tumor immune microenvironment(TIME).M2 macrophages are among the most extensively infiltrated immune cells in the TIME and are necessary for the g...Background:Pancreatic ductal adenocarcinoma(PDAC)has a rich and complex tumor immune microenvironment(TIME).M2 macrophages are among the most extensively infiltrated immune cells in the TIME and are necessary for the growth and migration of cancers.However,the mechanisms and targets mediating M2 macrophage infiltration in pancreatic cancer remain elusive.Methods:The M2 macrophage infiltration score of patients was assessed using the xCell algorithm.Using weighted gene co-expression network analysis(WGCNA),module genes associated with M2 macrophages were identified,and a predictive model was designed.The variations in immunological cell patterns,cancer mutations,and enrichment pathways between the cohorts with the high-and low-risk were examined.Additionally,the expression of FCGR3A and RNASE2,as well as their association with M2 macrophages were evaluated using the HPA,TNMplot,and GEPIA2 databases and verified by tissue immunofluorescence staining.Moreover,in vitro cell experiments were conducted,where FCGR3A was knocked down in pancreatic cancer cells using siRNA to analyze its effects on M2 macrophage infiltration,tumor proliferation,and metastasis.Results:The prognosis of patients in high-risk and low-risk groups was successfully distinguished using a prognostic risk score model of M2 macrophage-related genes(p=0.024).Between the high-and low-risk cohorts,there have been notable variations in immune cell infiltration patterns,tumor mutations,and biological functions.The risk score was linked to the manifestation of prevalent immunological checkpoints,immunological scores,and stroma values(all p<0.05).In vitro experiments and tissue immunofluorescence staining revealed that FCGR3A can promote the infiltration or polarization of M2 macrophages and enhance tumor proliferation and migration.Conclusions:In this study,an M2 macrophage-related pancreatic cancer risk score model was established,and found that FCGR3A was correlated with tumor formation,metastasis,and M2 macrophage infiltration.展开更多
目的:通过生物信息学分析以及细胞生物学实验研究角蛋白6A(KRT6A)对胰腺导管腺癌(PDAC)诊断、预后判断、免疫微环境以及PDAC细胞PANC1增殖、凋亡等生物学行为的影响。方法:通过GEPIA平台整合TCGA(The Cancer Genome Atlas)数据库与GTEx(...目的:通过生物信息学分析以及细胞生物学实验研究角蛋白6A(KRT6A)对胰腺导管腺癌(PDAC)诊断、预后判断、免疫微环境以及PDAC细胞PANC1增殖、凋亡等生物学行为的影响。方法:通过GEPIA平台整合TCGA(The Cancer Genome Atlas)数据库与GTEx(Genotype-Tissue)数据库中的数据,分析KTRT6A在PDAC组织中的表达情况,并通过CIBERSORT工具分析KRT6A表达与PDAC组织中免疫细胞浸润的关系,然后通过GSEA方法研究与KRT6A基因表达相关的肿瘤信号通路。选取长海医院病理科保存的60例PDAC组织与癌旁组织标本进行免疫组化分析,验证KRT6A在肿瘤组织中表达情况;通过干扰RNA敲减PANC1细胞中KRT6A的表达,采用CCK-8实验以及流式细胞术检测敲减KRT6A对细胞的增殖、凋亡的影响。结果:利用TCGA与GTEx数据库数据分析发现,KRT6A在人PDAC组织中呈高表达,且与患者较差的生存期存在关联(P=0.015)。利用CIBERSORT软件以及GSEA分析发现,KRT6A高表达的PDAC组织中M2型巨噬细胞浸润性升高(P=0.034),且与Wnt通路(NES:1.7359272,P<0.05)、磷酸戊糖途径(PPP)(NES:1.5613053,P<0.05)等信号通路上调有关联(P<0.05或P<0.01);免疫组化结果进一步验证了KRT6A在PDAC组织中呈高表达(P<0.001)。增殖和凋亡实验发现,干扰KRT6A能够显著抑制PANC1细胞的增殖(P<0.05)以及凋亡(P<0.001)。结论:KRT6A在人PDAC组织中呈高表达,敲减其表达能够抑制PANC1细胞的增殖和凋亡,具有作为PDAC诊断与预后判断新靶标的潜力。展开更多
文摘Pancreatic cancer has a dismal prognosis due to late detection and lack of efficient therapies.The Kirsten rat sarcoma virus(KRAS)oncogene is mutated in up to 90%of all pancreatic ductal adenocarcinomas(PDACs)and constitutes an attractive target for therapy.However,the most common KRAS mutations in PDAC are G12D(44%),G12V(34%)and G12R(20%)that are not amenable to treatment by KRAS G12C-directed cysteine-reactive KRAS inhibitors such as Sotorasib and Adagrasib that exhibit clinical efficacy in lung cancer.KRAS G12C mutant pancreatic cancer has been treated with Sotorasib but this mutation is detected only in 2%–3%of PDAC.Recently,the KRAS G12D-directed MRTX1133 inhibitor has entered clinical trials and more of such inhibitors are in development.The other KRAS mutations may be targeted indirectly via inhibition of the cognate guanosine exchange factor(GEF)Son of Sevenless 1 that drives KRAS.These agents seem to provide the means to target the most frequent KRAS mutations in PDAC and to improve patient outcomes.
文摘Background:Pancreatic ductal adenocarcinoma(PDAC)has a rich and complex tumor immune microenvironment(TIME).M2 macrophages are among the most extensively infiltrated immune cells in the TIME and are necessary for the growth and migration of cancers.However,the mechanisms and targets mediating M2 macrophage infiltration in pancreatic cancer remain elusive.Methods:The M2 macrophage infiltration score of patients was assessed using the xCell algorithm.Using weighted gene co-expression network analysis(WGCNA),module genes associated with M2 macrophages were identified,and a predictive model was designed.The variations in immunological cell patterns,cancer mutations,and enrichment pathways between the cohorts with the high-and low-risk were examined.Additionally,the expression of FCGR3A and RNASE2,as well as their association with M2 macrophages were evaluated using the HPA,TNMplot,and GEPIA2 databases and verified by tissue immunofluorescence staining.Moreover,in vitro cell experiments were conducted,where FCGR3A was knocked down in pancreatic cancer cells using siRNA to analyze its effects on M2 macrophage infiltration,tumor proliferation,and metastasis.Results:The prognosis of patients in high-risk and low-risk groups was successfully distinguished using a prognostic risk score model of M2 macrophage-related genes(p=0.024).Between the high-and low-risk cohorts,there have been notable variations in immune cell infiltration patterns,tumor mutations,and biological functions.The risk score was linked to the manifestation of prevalent immunological checkpoints,immunological scores,and stroma values(all p<0.05).In vitro experiments and tissue immunofluorescence staining revealed that FCGR3A can promote the infiltration or polarization of M2 macrophages and enhance tumor proliferation and migration.Conclusions:In this study,an M2 macrophage-related pancreatic cancer risk score model was established,and found that FCGR3A was correlated with tumor formation,metastasis,and M2 macrophage infiltration.
文摘目的:通过生物信息学分析以及细胞生物学实验研究角蛋白6A(KRT6A)对胰腺导管腺癌(PDAC)诊断、预后判断、免疫微环境以及PDAC细胞PANC1增殖、凋亡等生物学行为的影响。方法:通过GEPIA平台整合TCGA(The Cancer Genome Atlas)数据库与GTEx(Genotype-Tissue)数据库中的数据,分析KTRT6A在PDAC组织中的表达情况,并通过CIBERSORT工具分析KRT6A表达与PDAC组织中免疫细胞浸润的关系,然后通过GSEA方法研究与KRT6A基因表达相关的肿瘤信号通路。选取长海医院病理科保存的60例PDAC组织与癌旁组织标本进行免疫组化分析,验证KRT6A在肿瘤组织中表达情况;通过干扰RNA敲减PANC1细胞中KRT6A的表达,采用CCK-8实验以及流式细胞术检测敲减KRT6A对细胞的增殖、凋亡的影响。结果:利用TCGA与GTEx数据库数据分析发现,KRT6A在人PDAC组织中呈高表达,且与患者较差的生存期存在关联(P=0.015)。利用CIBERSORT软件以及GSEA分析发现,KRT6A高表达的PDAC组织中M2型巨噬细胞浸润性升高(P=0.034),且与Wnt通路(NES:1.7359272,P<0.05)、磷酸戊糖途径(PPP)(NES:1.5613053,P<0.05)等信号通路上调有关联(P<0.05或P<0.01);免疫组化结果进一步验证了KRT6A在PDAC组织中呈高表达(P<0.001)。增殖和凋亡实验发现,干扰KRT6A能够显著抑制PANC1细胞的增殖(P<0.05)以及凋亡(P<0.001)。结论:KRT6A在人PDAC组织中呈高表达,敲减其表达能够抑制PANC1细胞的增殖和凋亡,具有作为PDAC诊断与预后判断新靶标的潜力。
