摘要
目的观察体外乏氧条件下胰腺癌BXPC-3细胞中低氧诱导因子一2（HIF-2）和环氧合酶一2（cOX-2）的表达，探讨HIF-2在低氧条件下对胰腺癌细胞COx-2信号通路的调控作用。方法以CoCl。化学低氧法诱导肿瘤低氧状态，采用RT—PCR及Westernblot方法分别检测低氧微环境下HIF-2、COX-2mRNA和蛋白的表达。采用化学合成小干扰RNA（siRNA），转染BXPC-3细胞，进一步观察转染后HIF-2沉默对cOX-2信号通路的影响。结果在低氧条件下，BXPC-3细胞HIF-2mRNA表达稳定，与常氧组比较，低氧8、24h组HIF-2、cox_2mRNA表达均显著升高，差异有显著意义（F=529．49、63．99，q=8．99～46．02，P〈0．05），且低氧24h组高于低氧8h组（q=20．40、6．97，P〈0．05）；siRNA转染BXPC-3细胞后能够显著下调HIF-2、COX-2蛋白表达（F=243．98、63．99，q=8．99～30．69，P〈0．05）。结论低氧促使胰腺癌BXPC-3细胞HIF-2、COX-2mRNA和蛋白水平表达升高，C0x-2信号通路可能与HIF-2信号通路存在相互联系，并且共同促进胰腺癌的恶性进展。
Objective To investigate the expression of hypoxia-indueible factor-2 （HIF-2） and eyclooxygenase 2 （COX- 2） in human pancreatic cancer BXPC-3 cells under hypoxia, and observe the effect of HIF-2 in the regulation of COX-2 expression in the cancer BXPC-3 cells under hypoxic conditions. Methods Tumor hypoxia was induced by CoCl2 chemical hypoxia method, the expressions of HIF-2 and COX-2 in hypoxia microenvironment were respectively detected by RT-PCR and Western blot. RNA （siRNA） was chemically synthesized and transfected into BXPC-3 cells. The effect of HIF-2, after transfection, on COX-2 signal channel was observed. Results Under the condition of hypoxia, the expression of HIF-2 mRNA in BXPC-3 cells was stable, and compared with the normal oxygen group, in 8 h-and 24 h-hypoxia group, the expressions of HIF-2 mRNA and COX-2 mRNA were significantly higher, the differences were significant （F = 529.49,63.99 q = 8.99 -- 46.02 P 〈 0.05）, and that in 24 h-hypoxia group were higher than the 8 h-hypoxia （q =20.40,6.97 ;P〈0.05）. siRNA, after transfection with BXPC-3 cells, could significant- ly down-regulate the expressions of HIF-2 and COX-2 （F=243.98,63.99;q=8.99-30.69;P〈0.05）. Conclusion Hypoxia re- sults in the elevation of expressions of HIF-2, COX-2 mRNA and protein in BXPC-3 cells of pancreatic cancer. Signaling pathways COX-2 and HIF-2 are probably interconnected and leading to malignant progression of pancreatic cancer.
出处
《青岛大学医学院学报》
CAS
2015年第1期23-25,28,共4页
Acta Academiae Medicinae Qingdao Universitatis
基金
山东省高等学校科技计划项目(J12LL60)
