The overexpression of GRASP might inhibit cell proliferation and invasion in hepatocellular carcinoma

• Published in: Journal of cellular physiology 2019-09, Vol.234 (9), p.16215-16225
• Main Authors: Cheng, Yang, Yin, Baobing, Hou, Tianlu, Chen, Tianyang, Ping, Jian
• Format: Article
• Language: English
• Subjects: Bioinformatics; Bisulfite; Cancer; Cell growth; Cell proliferation; DNA methylation; Gene expression; Gene sequencing; Genes; Genomes; GRASP overexpression; Hepatocellular carcinoma; Liver cancer; Medical prognosis; Methylation; methylation subtypes; mRNA; Polymerase chain reaction; Prognosis; prognosis survival related model; proliferation; risk score; Survival
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.28285

This study aimed to validate the methylation of key genes in hepatocellular carcinoma (HCC) screened by bioinformatics analysis and explore whether they affected HCC cell proliferation, migration, and invasion. Using The Cancer Genome Atlas (TCGA) database, HCC‐related differentially methylated positions (DMPs) were screened, genes corresponding to DMPs were selected, and prognosis‐related genes were identified. A representative DMP was used to divide the DMPs into hyper‐ and hypomethylated groups. Expression of key genes in cell lines was detected using quantitative real‐time polymerase chain reaction and western blot analysis. After treatment of HepG2 cells with 5‐Aza‐2′‐deoxycytidine (5‐Aza‐DC), gene expression was observed. Bisulfite sequencing PCR assay was used to detect methylation frequency. Overexpressed GRASP lentiviral vectors were constructed to analyze their influence on cell proliferation, migration, and invasion using cell counting kit‐8 and transwell assays. Forty‐three HCC prognosis‐related genes were screened using the TCGA database. cg00249511 (SCT) was used to divide the DMPs into hyper‐ and hypomethylated groups, distinguishing between high‐ and low‐risk samples. The prognosis survival model constructed using 12 genes revealed the prognosis type. GRASP messenger RNA was downregulated in HepG2 and upregulated after 5‐Aza‐DC treatment. In HCC tissues, methylation frequency of GRASP was upregulated. GRASP overexpression inhibited HepG2 cell proliferation, invasion, and G‐CSFR expression. Thus, GRASP might be a prognosis‐related gene controlled by methylation. GRASP was downregulated in HepG2 cells, and upregulated after 5‐Aza‐2′‐deoxycytidine (5‐Aza‐DC) treatment. GRASP overexpression could inhibit the proliferative activity and invasion of hepatocellular carcinoma (HCC) cells. GRASP overexpression could significantly suppress the expression of G‐CSFR.