Effect Of CHFR CpG Island Methylation Status On Proliferation And Apoptosis In Eca109 Cell

Cell cycle checkpoint is the key transition point to ensure the evolution of the cell cycle, CHFR (checkpoint with FHA and ring finger) was first founded action on the mitosis prophase. It's one of most important tumor suppressor gene. When mitosis was under stress, CHFR expression could delay the chromosome gathered and stop cells in the prophase of mitosis. In normal tissue CHFR widely express. Its structure and function are very conservative. Research indicates that, many kinds of tumor tissue expression is missing, such as colon cancer, leukemia, gastric cancer, lung cancer and so on. The methylation of the CpG island is the most important mechanism to the missing or silence of the CHFR expression. As one of the most important way of the tumor-suppressor gene inactivation, the methylation of gene promoter plays a key role in the development of tumor. In recent years, it has become the research focus. Esophageal cancer is one of the most common human malignant tumors, a serious threat to human health and life. Especially inland is high occurrence area and squamous cell carcinoma is commonly seen. But in recent years esophageal adenocarcinoma is commonly and incidence is on the rise. There are few reports about CHFR gene research in esophageal cancer. To research the relationship between esophageal cancer and CHFR gene, we study the effect of CHFR CpG island methylation status on proliferation and apoptosis in Eca109 cell line. We look forward to exploring the pathogenesis of esophageal cancer and searching for a new target to cancer treatment.Objective: To study the effect of CHFR CpG island methylation status on proliferation and apoptosis in Eca109 cell line.Methods:1. The Eca109 cell was cultured and treated by 2,5,10μmol/L concentrations of 5-Aza-2'-deoxycytidine.2. MSP analysis was evaluated for the CpG island methylation status of the CHFR gene.3. RT-PCR was used to detect the semi-quantitative of CHFR mRNA expression.4. MTT were used to determine the proliferation in Eca109 cell treated by different concentrations of 5-Aza-CdR.5. Flow cytometer were used to determine the apoptosis in Eca109 cell treated by different concentrations of 5-Aza-CdR 24 hours. Results:1. In untreated Eca109 cell CHFR CpG island was hypermethylated, methylated CHFR gene was demethylated partly after treated with 5-Aza-CdR.2. No expression of CHFR mRNA was found in untreated Eca109 cell, but the relative expression of CHFR mRNA after treated by 5-Aza-CdR at concentrations of 2.5, and 10μmol/L were 0.174±0.010,0.221±0.013,0.356±0.014, the difference was statistically significant (P<0.05).3. Different concentrations of 5-Aza-CdR treatment inhibited proliferation in Eca109 cell, the difference was statistically significant (P<0.05). Within a certain range, Eca109 cell growth inhibition rate increased with the processing of drug concentration and prolonged action time.4. Untreated and respectively 2,5, and 10μmol/L of 5-Aza-CdR treatment Eca109 cell apoptosis rates were:1.83±0.41%,7.46±1.46%,16.27±1.61%, 25.29±2.25%, the difference was statistically significant (P<0.01).Conclusion:Esophageal cancer Eca109 cell after 5-Aza-CdR treatment, CHFR CpG island can be demethylated partly, CHFR mRNA express, inhibit cell proliferation and promote apoptosis.