Mifepristone Regulating The H19 Methylation Inhibition Of Endometrial Cancer Cell Migration

Background and ObjectiveEndometrial carcinoma (EnCa) is a common type of female genital tract malignancy. The invasion and migration ability have been known as the important factors to influence the EnCa’s outcome, relapse and mortality rate. People pay more and more attention to researching the source of EnCa’s invasive and migration ability. Mifepristone is a kind of progestogen and glucocorticoids antagonists. A lots of researches detected that Mifepristone have antitumor effect, but its mechanism is unclear. Researched results shew that H19 in endometrial carcinoma tissues was characterized by high expression, The H19 by let7/high mobility group protein A2 (HMGA2) promoting the Epithelia to mesenchymal transition (EMT), metformin can inhibits tumor cell migration via promoting H19 methylation and downregulating the expression of H19.Thus speculate that mifepristone may also through regulating the H19 methylation to influence of endometrial cancer cell migration. In this study, we take the human endometrial carcinoma cell line Ishikawa cells as the research object, to investigate the effect and mechanism of mifepristone on human endometrial carcinoma cells.Methods1. Human endometrial carcinoma cell line Ishikawa cells were cultured in vitro Jshikawa cells be divided into 5 mg/L mifepristone treatment group,10 mg/L mifepristone group and the control group,Wound healing assay was applied to detect the migration of Ishikawa cells.2. Ishikawa cells be divided into 2.5 mg/L mifepristone treatment group,5 mg/L mifepristone group,10 mg/L mifepristone treatment group and control group, Dosing treatment for 24h,48h and 72 h. RT-PCR and methylation-specific PCR, (MSP) was used to detect the expression of H19 mRNA and the DNA methylation.3. Ishikawa cells be divided into 2.5 mg/L mifepristone treatment group,5 mg/L mifepristone group,10 mg/L mifepristone treatment group and control group, Dosing treatment for 24h> 48 hand 72 h. Western blotting was used to detect the expression of HMGA2 and EMT related protein.Results1. When treated by mifepristone for 48 hours, the Scratch widths of the the control group、the 5 mg/L mifepristone treatment groups and the 10 mg/L mifepristone treatment group were 125.24±1.96、140.42±2.03、149.58±2.05; Compared with the control groups, with statistical differences (P<0.05).When treated by mifepristone for 72 hours, the Scratch widths of the the control group、the 5 mg/L mifepristone treatment groups and the 10 mg/L mifepristone treatment group were 103.64±2.00、128.59±2.04、143.42±1.09; Compared with the control groups, with statistical differences (P<0.05).2. Iskikawa cells, with the increase of the time and concentration, the inhibition effect of mifepristone for H19 mRNA expression increases, and the promoting effect for H19 gene promoter region methylation increases.3. Iskikawa cells, with the increase of the time and concentration, the inhibition effect for HMGA2 expression increases, then E-cadherin expression rise with the increase of concentration, the expression of Vimentin decreased with the increase of concentration.ConclusionMifepristone could inhibits tumor cell migration, partly by downregulating H19 mRNA via H19 gene methylation, and then downregulating HMGA2 and Vimentin expression, increasing E - cadherin expression.