CAJ | 학술논문

目的研究低氧对胰腺癌乙酰肝素酶(Hpa)表达的影响及与肿瘤细胞侵袭力的关系.方法体外低氧培养细胞,观察低氧在不同时间胰腺导管癌细胞株BxPC-3细胞Hpa mRNA和蛋白表达的变化;以Tranwell侵袭小室实验观察低氧及应用反义寡核苷酸(AS-ODN)阻断Hpa表达后肿瘤细胞侵袭力的变化,明确低氧促进肿瘤细胞侵袭力增加是否与Hpa有关.结果在常氧条件下,BxPC-3细胞Hpa mRNA和蛋白即有低量表达.在低氧条件下,Hpa mRNA和蛋白表达在3 h受到轻度抑制,在6 h、12 h、24 h和48 h表达明显升高,且mRNA和蛋白的表达水平一致.预先以Hpa AS-ODN序列阻断Hpa的表达,则低氧不能诱导Hpa mRNA和蛋白表达上调.低氧使穿过Transwell小室的BxPC-3细胞数量增加了96.2%(P<0.01),Hpa AS-ODN(400 nmol/L)对低氧诱导的侵袭细胞的抑制率为37.2%,差异有统计学意义(P<0.05).结论低氧可通过上调胰腺癌BxPC-3细胞Hpa mRNA和蛋白的表达,提高肿瘤细胞侵袭力.
목적 연구저양대이선암을선간소매(Hpa)표체적영향급여종류세포침습력적관계.방법 체외저양배양세포,관찰저양재불동시간이선도관암세포주BxPC-3세포Hpa mRNA화단백표체적변화;이Tranwell침습소실실험관찰저양급응용반의과핵감산(AS-ODN)조단Hpa표체후종류세포침습력적변화,명학저양촉진종류세포침습력증가시부여Hpa유관.결과 재상양조건하,BxPC-3세포Hpa mRNA화단백즉유저량표체.재저양조건하,Hpa mRNA화단백표체재3 h수도경도억제,재6 h、12 h、24 h화48 h표체명현승고,차mRNA화단백적표체수평일치.예선이Hpa AS-ODN서렬조단Hpa적표체,칙저양불능유도Hpa mRNA화단백표체상조.저양사천과Transwell소실적BxPC-3세포수량증가료96.2%(P<0.01),Hpa AS-ODN(400 nmol/L)대저양유도적침습세포적억제솔위37.2%,차이유통계학의의(P<0.05).결론 저양가통과상조이선암BxPC-3세포Hpa mRNA화단백적표체,제고종류세포침습력.
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.