HNF-1a 通过其靶基因 PKLR 促进胰腺癌的生长和抗凋亡。

HNF-1a promotes pancreatic cancer growth and apoptosis resistance via its target gene PKLR.

机构信息

Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Shanghai 200032, China.

Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China.

出版信息

Acta Biochim Biophys Sin (Shanghai). 2020 Mar 18;52(3):241-250. doi: 10.1093/abbs/gmz169.

DOI:10.1093/abbs/gmz169

PMID:32072180

Abstract

Pancreatic ductal adenocarcinoma is one of the deadliest malignant tumors, and many genes play important roles in its development. The hepatocyte nuclear factor-1a (HNF-1a) gene encodes HNF-1a, which is a transcriptional activator. HNF-1a regulates the tissue-specific expression of multiple genes, especially in pancreatic islet cells and in the liver. However, the role of the HNF-1a gene in the development of pancreatic cancer is still unclear. Here, we used immunohistochemical staining and real-time PCR to analyze HNF-1a expression in pancreatic cancer tissue. Stable cell lines with HNF-1a knockdown or overexpression were established to analyze the role of HNF-1a in pancreatic cancer cell proliferation and apoptosis by colony formation assay and flow cytometry. We also analyzed the L-type pyruvate kinase (PKLR) promoter sequence to identify the regulatory effect of HNF-1a on PKLR transcription and confirmed the HNF-1a binding site in the PKLR promoter via a chromatin immunoprecipitation assay. HNF-1a was found to be overexpressed in pancreatic cancer and promoted proliferation while inhibiting apoptosis in pancreatic cancer cells. PKLR was identified as the downstream target gene of HNF-1a and binding of HNF-1a at two sites in PKLR (-1931/-1926 and -966/-961) regulated PKLR transcription. In conclusion, HNF-1a is overexpressed in pancreatic cancer, and the transcription factor HNF-1a can promote pancreatic cancer growth and apoptosis resistance via its target gene PKLR.

摘要

胰腺导管腺癌是最致命的恶性肿瘤之一，许多基因在其发展中发挥重要作用。肝细胞核因子-1a（HNF-1a）基因编码转录激活因子 HNF-1a。HNF-1a 调节多个基因的组织特异性表达，特别是在胰腺胰岛细胞和肝脏中。然而，HNF-1a 基因在胰腺癌发展中的作用尚不清楚。在这里，我们使用免疫组织化学染色和实时 PCR 分析了胰腺癌组织中 HNF-1a 的表达。通过集落形成实验和流式细胞术建立了 HNF-1a 敲低或过表达的稳定细胞系，分析了 HNF-1a 在胰腺癌细胞增殖和凋亡中的作用。我们还分析了 L 型丙酮酸激酶（PKLR）启动子序列，以确定 HNF-1a 对 PKLR 转录的调节作用，并通过染色质免疫沉淀实验证实了 HNF-1a 在 PKLR 启动子上的结合位点。发现 HNF-1a 在胰腺癌中过表达，促进了胰腺癌细胞的增殖，同时抑制了细胞凋亡。鉴定出 PKLR 是 HNF-1a 的下游靶基因，HNF-1a 在 PKLR（-1931/-1926 和 -966/-961）的两个位点结合调节 PKLR 转录。总之，HNF-1a 在胰腺癌中过表达，转录因子 HNF-1a 可以通过其靶基因 PKLR 促进胰腺癌的生长和抗凋亡。