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

Abstract

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.

Early detection and prognosis evaluation for hepatocellular carcinoma by circulating tumour DNA methylation: A multicentre cohort study

“…Finally, we identified 183 common hypermethylated DMRs in cancer samples compared to all other controls, irrespective of the methodology (Figure 2A and B, see Supplementary Methods in Supplementary Material). Several of these markers have been previously reported to be relevant in tumourigenesis 38–40 (Figure 2C and D), confirming the efficacy of the marker selection. After curation of the genomic location, gene annotation and literature‐based evidence, we assessed the analytical accuracy of the candidate markers by qMSP in an additional 20 HCC tumours, 20 peritumoural tissues, and 10 WBCs from healthy participants.…”

Effect of CELSR3 on the Cell Cycle and Apoptosis of Hepatocellular Carcinoma Cells

“…Previous studies have suggested that the methylation of genes plays an important role in hepatocarcinogenesis. For instance, GRASP overexpression controlled by methylation would significantly suppress the proliferation and invasion of HCC cells [42]. Our study demonstrated that the methylation of CELSR3 was decreased in HCC and negatively correlated with the expression of CELSR3.…”

Identification of potential prognostic genes associated with the tumor microenvironment in chromophobe renal cell carcinoma based on weighted gene co-expression network analysis

“…It is popularly referred that cancer cells undergo mitosis in order to reproduce. Cheng et al demonstrated that over-expression of GRASP had an inhibitory effect on the activity of HepG2 cells, suggesting that GRASP may decrease the proliferation of HCC cells [41]. Shen et al originally proved that expression of GRAP is primarily regulated by DNA methylation [42; 43].…”