Original Article
Research on the expression and regulation of ASPP2 and its methylationin human gastric carcinoma cell
Jianyan Tang, Dengqiu Zhao, Yefeng Wu, Longxiang Zhou
Published 2019-06-25
Cite as Chin J Endocr Surg, 2019, 13(3): 208-213. DOI: 10.3760/cma.j.issn.1674-6090.2019.03.008
Abstract
ObjectiveTo investigate the relationship between the expression of ASPP2 mRNA and the methylation of ASPP2 gene in gastric cancer cells, to observe the inhibitory effect of 5-Aza-CdR on the growth of gastric cancer cells, to observe the effect of demethylation on the expression of ASPP2 mRNA and the methylation of ASPP2 gene in gastric cancer cells, and to explore the molecular mechanism of gastric cancer.
MethodsReal-time PCR was used to detect the expression of ASPP2 mRNA in two gastric cancer cells and normal gastric epithelial cells. BSP was used to detect the methylation of ASPP2 gene in two gastric cancer cells and normal gastric epithelial cells. CCK-8 was used to detect the growth inhibition rate of gastric cancer cells treated with 5-Aza-CdR of different concentrations, then they were used to detect expression of ASPP2 mRNA and the methylation of ASPP2 gene in gastric cancer cells again after the demethylation.
Results① The expression of ASPP2 mRNA in MKN-45 cells was significantly lower than that in GES-1 cells (P<0.01) . The expression of ASPP2 mRNA in MGC-803 cells was significantly lower than that in GES-1 cells (P<0.01) . There was no significant difference in MGC-803 cells and MKN-45 cells (P>0.05) . ② The methylation rate of ASPP2 in MKN-45 cells was significantly higher than that in GES-1 cells (P<0.01) . The methylation rate of ASPP2 in MGC-803 cells was not significantly different from that in GES-1 cells (P>0.05) . The methylation rate of ASPP2 in MKN-45 cells was significantly higher than that in MGC-803 cells (P<0.01) . ③ At the same time, the growth inhibition rate of each 5-Aza-CdR concentration group increased as the drug concentration depended.4.The expression of ASPP2 mRNA in MKN-45 cells was significantly higher than that before treatment (P<0.01) , the expression of ASPP2 mRNA in MGC-803 cells was not significantly different from that before treatment (P>0.05) . The methylation rate of ASPP2 in MKN-45 cells was significantly lower than that before treatment (P<0.01) .The methylation rate of ASPP2 in MGC-803 cells was not significantly different from that before treatment (P>0.05) .
Conclusion① Abnormal hypermethylation of ASPP2 gene in MKN-45 cells may be a molecular mechanism of decreased ASPP2 mRNA expression.② 5-Aza-CdR can inhibit the growth of MKN-45 and MGC-803 cells, and it can enhance the expression of ASPP2 mRNA in MKN-45 cells. Reversal of methylation in the promoter region of ASPP2 gene is the possible mechanism. ③ Abnormal hypermethylation of the promoter region of ASPP2 gene may lead to silencing of mRNA expression that may be associated with gastric cancer.
Key words:
ASPP2; DNA methylation; Gastric carcinoma; 5-Aza-CdR
Contributor Information
Jianyan Tang
Department of Radiology, Shanghai Fengxian District Central Hospital, Shanghai 201499, China
Dengqiu Zhao
Department of Hepatobiliary Surgery, Shanghai University of Medicine &
Health Sciences Affiliated Zhoupu Hospital, Shanghai 201318, China
Yefeng Wu
Department of General Surgery, the Sixth Affiliated People’s Hospital of Jinshan Branch, Shanghai 201599, China
Longxiang Zhou
Department of General Surgery, the Sixth Affiliated People’s Hospital of Jinshan Branch, Shanghai 201599, China
