The Extract Of Centipede Effects On Human Hepatic Cells Methylation

Objective:Discuss the different of the genome-wide methylation research between High and low degree of malignant liver cancer cells which treated with the Centipede extraction(CE) with the whole genome DNA methylation chip (Ⅲlumina infinium human methylation450).Methods:The CE were prepared by methods of following precipitation with acetone. The cellular proliferation was assessed with MTS in low degree of malignant liver cancer cells HepG2and High degree of malignant liver cancer cells HCCLM3used with the drug. Calculate the IC50alue concentration. CE sequential processing with1/2IC50con--centration of three types of cells in3d,5μL concentration5-Azacytudine and blank comparison group, Genome-wide methylation chip are used to observe drugs on human hepatocellular carcinoma HepG2, HCCLM3cells genome-wide methylation spectrum.Using BeadStudio Methy-lation Module v3.2software to analyze data and the degree of Methylation differences sites for screening.Result:(1) CE effect HepG2and HCCLM3ells after48h, MTS results show that the extracts are inhibition of cell growth, the centipede extract different drug concentrations under the action of the cell inhibition rate compared with control group (P<0.05). Calculation of drug intervention48h HepG2and HCCLM3IC50value concentration of0.613mg/mL and0.435mg/mL.(2)①Genome-wide chip shows HepG2blank group and HCCLM3blank group of481known genetic differences between methylation site, the highest methylation site located in GSTA4gene, methylation site is located at the lowest RIN4genes;②centipede in treatment group HepG2and HCCLM3centipede group (HepG2-HCCLM3CE) have845known genetic differences between methylation site, the highest methylation site is located in the GJB2gene, methylation site is located in the lowest IRS4genes;③5-Azacytudine treatment group, there are628known genetic difference in methylation site between HepG2and HCCLM3group(HepG2-HCCLM35AZ), the highest methylation site located in EPHB4gene, methylation site is located in the lowest IRS4genes;④HepG2-HCCLM3CE and HepG2-HCCLM3have438known genetic differences between methylation site, the highest methylation site is located in the GSTA4gene, methylation site is located in the lowest IRS4genes; HepG2-HCCLM35AZ and HepG2-HCCLM3have291known genetic differences between methylation site, the highest methylation site is located in the GJB2gene, methylation site is located in the lowest IRS4genes.Conclusion:(1) The centipede extract can inhibit HCCLM3and HepG2cells in vitro,IC50concentration is0.613mg/mL and0.435mg/mL.(2) Genome-wide chip shows HepG2blank group and HCCLM3blank group of481knowngenetic differences between methylation site, the high methylation site189, the low methylation site292;(3) The differences of CE intervention can affect HepG2-HCCLM3methylation site,but at variance with5-Azacytudine.