Chen Peng, Pan Meng, Shen Zhengchao, Yang Yuquan, Wang Xiaoming
Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Wannan Medical College, Wuhu, Anhui 241000, P.R. China.
Department of Hepatobiliary Surgery, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui 241001, P.R. China.
J Cancer. 2024 Feb 17;15(7):2033-2044. doi: 10.7150/jca.90689. eCollection 2024.
It is crucial to probe into the biological effect and mechanism of miRNA-485-5p regulating keratin 17 (KRT17) in pancreatic cancer (PC) to understand its pathogenesis and identify potential biological targets. The bioinformatics means were used to evaluate the clinical significance of KRT17 expression in the Cancer Genome Atlas (TCGA) database. TargetScan database analysis in conjunction with dual luciferase and RNA Immunoprecipitation (RIP) experiments was used to probe the interaction relationship of miRNA-485-5p with KRT17. The expression of miRNA-485-5p and KRT17 in PC tissue and cancer cell lines was detected by Q-PCR paired with western blot assay. The biological function of miRNA-485-5p in regulating KRT17 was investigated in the PC cell line via gene silencing/overexpression technique. A western blot experiment was utilized to investigate the regulatory effect of KRT17 on cell cycle-related proteins and the FAK/Src/ERK signal pathway. The level of KRT17 was increased in PC tissues and this significantly decreased the survival rate of PC patients. TargetScan in combination with dual luciferase and RIP experiments verified the miRNA-485-5p target KRT17. The expression of KRT17 was high in the PC cell line, although the expression of miRNA-485-5p was low. Silencing KRT17 or overexpression of miRNA-485-5p significantly inhibited PC cell viability, proliferation, invasion, and colony formation, while promoting apoptosis. Overexpression of KRT17 drastically reversed the function of miRNA-485-5p. The silenced KRT17 remarkably downregulated the expression of cyclinD1, Cyclin Dependent Kinase 1 (CDK1), CDK2, Phospho-Focal Adhesion Kinase (p-FAK), p-Src, and p-ERK proteins in the PC cells. Generally, an essential signaling cascade of miRNA-485-5p/KRT17/FAK/Src/ERK influences the biological functions of PC cells.
探究miRNA - 485 - 5p调控胰腺癌（PC）中角蛋白17（KRT17）的生物学效应及机制，对于理解其发病机制和识别潜在生物学靶点至关重要。运用生物信息学方法评估癌症基因组图谱（TCGA）数据库中KRT17表达的临床意义。结合TargetScan数据库分析以及双荧光素酶和RNA免疫沉淀（RIP）实验，探究miRNA - 485 - 5p与KRT17的相互作用关系。通过Q - PCR结合蛋白质免疫印迹法检测PC组织和癌细胞系中miRNA - 485 - 5p和KRT17的表达。利用基因沉默/过表达技术在PC细胞系中研究miRNA - 485 - 5p调控KRT17的生物学功能。运用蛋白质免疫印迹实验研究KRT17对细胞周期相关蛋白以及FAK/Src/ERK信号通路的调控作用。PC组织中KRT17水平升高，这显著降低了PC患者的生存率。TargetScan结合双荧光素酶和RIP实验验证了miRNA - 485 - 5p靶向KRT17。PC细胞系中KRT17表达较高，而miRNA - 485 - 5p表达较低。沉默KRT17或过表达miRNA - 485 - 5p显著抑制PC细胞活力、增殖、侵袭和集落形成，同时促进细胞凋亡。KRT17的过表达显著逆转了miRNA - 485 - 5p的功能。沉默KRT17显著下调PC细胞中细胞周期蛋白D1、细胞周期蛋白依赖性激酶1（CDK1）、CDK2、磷酸化粘着斑激酶（p - FAK）、p - Src和p - ERK蛋白的表达。总体而言，miRNA - 485 - 5p/KRT17/FAK/Src/ERK这一关键信号级联影响PC细胞的生物学功能。
