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[颜面生殖器发育异常6基因对肝干细胞分化的调控作用]

[Regulatory effect of faciogenital dysplasia 6 gene on hepatic stem cell differentiation].

作者信息

Xue Q, Yang J F, Li B, He T C, Zhang B Q

机构信息

Department of Gastroenterology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.

Molecular Oncology Laboratory, the University of Chicago Medical Center, Chicago 60637, USA.

出版信息

Zhonghua Gan Zang Bing Za Zhi. 2017 Apr 20;25(4):268-272. doi: 10.3760/cma.j.issn.1007-3418.2017.04.007.

Abstract

To investigate the regulatory effect of faciogenital dysplasia 6 (FGD6) gene on hepatic stem cell differentiation. FGD6 gene was selected for the co-intervention of target sequence, the AdEasy system was used for the construction of adenovirus vector and the packaging and multiplication of the recombinant adenovirus vector pSES-FGD6-siRNA, and the HP14.5 cells were infected. Immunofluorescence assay was used to measure the expression of FGD6 protein in HP14.5 cells, quantitative real-time PCR was used to measure the mRNA expression of FGD6, alpha-fetoprotein (AFP), and albumin (Alb), and Western blot was used to measure the protein expression of FGD6, AFP, and Alb. The empty pSES-Ad-RFP adenovirus vector was constructed as control in each group. All data were expressed as x±s, and a one-way analysis of variance was performed. FGD6 protein was mainly expressed in the nucleus of HP14.5 cells. The pSES-FGD6-siRNA adenovirus vector was successfully constructed and it downregulated the expression of FGD6 gene and the mRNA and protein expression of AFP in HP14.5 cells and upregulated the mRNA and protein expression of Alb ( < 0.01). The inhibition of the expression of FGD6 gene in HP14.5 cells may differentiate HP14.5 cells into hepatocytes. Therefore, FGD6 gene plays an important role in the differentiation regulation of hepatic stem cells.

摘要

探讨颜面生殖器发育异常6(FGD6)基因对肝干细胞分化的调控作用。选择FGD6基因进行靶序列共干预,采用AdEasy系统构建腺病毒载体并包装、扩增重组腺病毒载体pSES-FGD6-siRNA,感染HP14.5细胞。采用免疫荧光法检测HP14.5细胞中FGD6蛋白的表达,采用定量实时PCR检测FGD6、甲胎蛋白(AFP)和白蛋白(Alb)的mRNA表达,采用蛋白质免疫印迹法检测FGD6、AFP和Alb的蛋白表达。每组构建空的pSES-Ad-RFP腺病毒载体作为对照。所有数据以x±s表示,进行单因素方差分析。FGD6蛋白主要表达于HP14.5细胞的细胞核中。成功构建了pSES-FGD6-siRNA腺病毒载体,其下调了HP14.5细胞中FGD6基因的表达以及AFP的mRNA和蛋白表达,并上调了Alb的mRNA和蛋白表达(<0.01)。抑制HP14.5细胞中FGD6基因的表达可能促使HP14.5细胞分化为肝细胞。因此,FGD6基因在肝干细胞的分化调控中起重要作用。

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