HTRA1 gene expression in gastric epithelial cells

AbstractObjectivesTo explore HtrA1 gene expression and its regulation in human gastric cancers.MethodsThe HtrA1 mRNA levels were examined by QPCR analysis and confirmed its expression with Northern blot analysis. The HtrA1 protein levels in all six gastric epithelial cell lines were investigated by Western blot analysis. Gene copy number was accessed and then sequenced the coding region from each mRNA in all six cell lines. The HtrA1 promoter region DNA methylation status was detected by using bisulfite sequencing analysis. Effect of decitabine and TSA on HTRA1 expression in gastric cancer cell line was determined by RTPCR.ResultsHIC analysis indicated that HtrA1 was highly expressed in normal epithelium, but dramatically down-regulated in gastric carcinoma tissues and variably expressed in tumor-adjacent tissues. HtrA1 gene expression was dramatically decreased in gastric carcinoma cells compared to non-tumorigenic counterparts. The HtrA1 gene loss in any of the 4 breast cancer cell lines was not detected. Total 14 CpGs in this region were all methylated in gastric cancer cells, whereas two normal cells, GES-1 and HFI-145, were having several unmethylated cytosines in this region. HtrA1 showed as ~Mr 44,000, Expression of HtrA1 protein was not observed in any of the four gastric cancer cell lines, BGC-823, MKN-45, SGC-7901and MKN-28. HtrA1 expression was observed in the HFI-145and GES-1 cell lines.ConclusionsThe epigenetic silencing for HtrA1 gene expression could provide a possible strategy for re-activating HtrA1 gene expression in gastric cancer cells, thus facilitating further investigation of HtrA1's role in chemotherapy