Effects of antisense oligodeoxynucleotide on expression of heparanase mRNA and invasion of pancreatic carcinoma in hypoxia / 中华内分泌外科杂志

ABSTRACT

Objective To study the regulation of heparanase expression by hypoxia and its correlation to the invasiveness of tumor cells. Methods BxPC-3 cells were cultured in hypoxia in vitro and the heparanase mRNA and protein expression were detected by reverse transcriptional polymerase reaction chains (RT-PCR) and western blot respectively. Matrigel invasion assay was used to observe the invasive abilities of tumor cells in hypoxia and in the status of heparanase was inhibited by antisense oligodeoxynucleotide (AS-ODN) targeting to the heparanase gene promoter. Results In normoxia, there was a relatively low level of heparanase mRNA and protein expression in cultured BxPC-3 cells. In hypoxia, heparanase expression, mRNA and protein which expressed consistently, were inhibited slightly at 3h and upregulated significantly at 6h, 12h, 24h and 48h. When the heparanase expression was inhibited by AS-ODN, the heparanase mRNA and protein maintained low in hypoxia, however, the nonsense oligodeoxynucleotide (NS-ODN) did not block upregulation of heparanase expression. In matrigel assay, after 48h incubation, number of BxPC-3 cells that penetrated the Matrigel-coated filter of transwell chamber was increased 96.2% in hypoxia (P<0.01), the Hpa AS-ODN (400 nmol/L) inhibited the invasive cells by 37.2% (P<0.05). Conclusions BxPC-3 cells invasion ability is enhanced by hypoxia through upregulation of heparanase mRNA and protein expression in BxPC-3 pancreatic cancer cell lines.