Effects Of The Recovered Expression Of Sfrp1 Gene On MGC803 Cell Line Proliferation And Migration Ability

Objective Gastric carcinoma is one of the most common malignant tumor, which caused the second cancer death rate in the world. Due to atypical early symptoms of gastric cancer, most patients are diagnosed with terminal cancer. However, patients with advanced gastric cancer 5 year survival rate is less than 10%.It is significant of searching for new therapeutic targets and chemotherapy drugs.The abnormal activation of Wnt signaling pathways is closely related to the development of tumor, and its antagonists silenced for epigenetic mechanisms can be found as a common event in tumorigenesis. The methylation of tumor suppressor genes can be reversed. We aim to investigate the effects of SFRP1 expression and biological characteristics of gastric cancer MGC803 cells induced by methyltransferase inhibitor 5-aza-2’ – deoxycytidine(5-Aza-d C),which may provide the experiment foundation of 5-Aza-d C applied in the clinical treatment of Gastric carcinoma.Methods1.We cultured gastric cancer MGC- 803 cells in vitro, take the logarithm cell randomly divided into blank control group, 1 umol/L 5-Aza-d C treatment group, 5umol/L 5-Aza-d C treatment group, 10umol/L 5-Aza-d C treatment group and 20umol/L 5-Aza-d C treatment group. 2. Using MTT method calculate the OD value of each group in 0 h、12 h、24 h,、48 h, Observing the ability of 5-Aza-d C to inhibit MGC- 803 cell proliferation, According to the MTT results map a growing curve inhibition, and preliminary screen a high inhibition rate. 3.Take the certain concentration of 5-Aza-d C respectively affect cell 0 h、12 h、24 h、48 h, The m RNA and ptotein expression of SFRP1 and β-catenin in gastric cancer cell line MGC-803 was respectively determined by using a real-time reverse transcription polymerase chain reaction(RT-PCR) assay and western blotting. 4. The cell scratch test was carried out to observe the migration ability of gastric cancer cell line MGC-803.Results 1. MTT result revealed that the proliferation of MGC-803 cells was significantly inhibited in a dose-dependent manner. 2. The result of RT-PCR showed that the expression of SFRP1 increased markedly and the expression of β-catenin decreased markedly, the difference was significant(p<0.05). There is negative correlation between the two genes in m RNA expression.3. The result of Western blot showed that the expression of SFRP1 increased markedly and the expression of β-catenin decreased markedly, the difference was significant(p<0.05). There is negative correlation between and the two genes in protein expression. 4.The cell scratch test exhibited: After 12 hours,the healing rate of MGC-803 cells of 5μmol/L group(47.8%±3.1%)and 10μmol/L group(18.2%±2.8%)was lower than blank group(54.5.0%±1.6%), the difference was statistically significant(P<0.05).After 24 hours later, The healing rate of MGC-803 cells of 5μmol/L group(82.7%±1.9%)and 10μmol/L group(45.5%±1.3%)was lower than blank group(91%±2.5%), the difference was statistically significant(P<0.05).Conclusion 1.5-Aza-d C can significantly inhibit the abilities of proliferation and migration of human gastric cancer cell line MGC-803,which possess the feature of time and the concentration dependence. 2.5-Aza-d C may increase SFRP1 gene expression by reversing SFRP1 gene methylation, which exert an effect on inhibiting tumor proliferation and migration.