KRT16在胰腺癌中的功能、临床意义及预后价值

The function,clinical significance and prognostic values of KRT16 in pancreatic carcinoma

目的:研究角蛋白16(keratin 16,KRT16)在胰腺癌细胞系SW1990中的细胞生物学功能。观察KRT16在胰腺癌组织中的表达,分析其表达水平与胰腺癌患者临床病理特征及预后的关系,探讨KRT16在胰腺癌中的临床意义及预后价值,并预测可能的相关机制。方法:分别采用EdU免疫荧光实验、Transwell实验、划痕实验、流式细胞术检测KRT16对SW1990细胞增殖、侵袭、迁移及凋亡的影响。收集并整理2016年01月至2018年12月期间在我院接受手术治疗的56例胰腺癌患者的组织标本及临床病例资料。采用免疫组织化学染色(immunohistochemistry, IHC)检测KRT16蛋白在胰腺癌组织中的表达,统计分析KRT16蛋白表达水平与胰腺癌患者临床病理特征及预后的关系。KRT16相互作用蛋白网络可通过STRING数据库获取,利用基因集富集分析(gene set enrichment analysis, GSEA)预测KRT16在胰腺癌中可能参与的信号通路。结果:细胞功能实验检测结果表明过表达KRT16促进胰腺癌细胞SW1990增殖、侵袭迁移并抑制其凋亡(P<0.001),同时,沉默KRT16则抑制胰腺癌细胞SW1990增殖、侵袭迁移并诱导其凋亡(P<0.001)。IHC检测结果显示KRT16在36例胰腺癌组织中高表达,在20例中低表达。KRT16蛋白表达水平、M分期是胰腺癌患者预后的独立危险因素(P<0.05)。Kaplan-Meier生存曲线结果显示:KRT16高表达组胰腺癌患者的生存时间显著低于KRT16低表达组患者(P<0.001)。STRING数据库结果表明:KRT16与KRT5等蛋白存在相互作用。GSEA研究结果显示:KRT16 mRNA高表达样本富集到VEGF信号通路、轴突导向等相关基因集(P<0.05)。结论:KRT16促进胰腺癌细胞增殖、侵袭及迁移。KRT16在胰腺癌中高表达并与胰腺癌的不良预后密切相关,KRT16可能通过调节KRT5等相互作用蛋白及调控VEGF等信号通路在胰腺癌中发挥功能。有望成为新的胰腺癌预后判断指标和治疗靶点。

Objective:To research the biological functions of KRT16 in SW1990 cell line.To observe the expression of KRT16 in pancreatic carcinoma tissues.And to analyze the relationship between KRT16 expression level and clinicopathologic feature,prognosis of pancreatic carcinoma patients.Moreover,to investigate the clinical significance and prognosis value of KRT16 in pancreatic carcinoma.And to explore possible related mechanisms.Methods:EdU immunofluorescence test,Transwell assay,scratch test and flow cytometry were used to detect the proliferation,invasion,migration and apoptosis of SW1990 cells,respectively.Meanwhile,56 cases of tissue samples and clinical case data from January 2016 to December 2018 were collected in our hospital.KRT16 protein expression in pancreatic carcinoma tissues was detected by immunohistochemistry(IHC).The correlation between KRT16 expression level and clinicopathologic feature,prognosis of pancreatic carcinoma patients were analyzed.The STRING database was used to analyze the network of proteins interacting with KRT16.Gene set enrichment analysis(GSEA)was used to predict the possible signal pathways of KRT16 in pancreatic carcinoma.Results:Cell function experiment results showed that overexpression KRT16 significantly enhanced SW1990 cell proliferation,invasion,migration and inhibited its apoptosis(P<0.001).However,interference KRT16 significantly inhibited SW1990 cell proliferation,invasion,migration and inhanced its apoptosis(P<0.001).IHC results showed that KRT16 protein was high expression in 36 cases of pancreatic carcinoma tissues and low expression in 20 cases.Then KRT16 protein expression level and M stage were independent risk factors affecting the prognosis of pancreatic carcinoma patients(P<0.05).Kaplan-Meier survival curves revealed that pancreatic carcinoma patients with high KRT16 levels had shorter survival(P<0.001).STRING database showed that there was an interaction between KRT16 and KRT5 and so on.GSEA research results showed that the KRT16 mRNA high expression samples were enriched into VEGF signaling pathway,axon guidance and so on(P<0.05).Conclusion:KRT16 promotes the proliferation,invasion and migration ability of SW1990 cell.Meanwhile,the high expression of KRT16 may be predict an unfavorable prognosis in pancreatic carcinoma patients.Moreover,it plays an important role in pancreatic carcinoma by regulating interacting proteins KRT5 and activating VEGF signaling pathways.And it may serves as a novel prognostic factor and therapeutic target of pancreatic carcinoma.