Hua Keqin, Din Jingxin, Cao Qi, Feng Weiwei, Zhang Ying, Yao Liangqin, Huang Yan, Zhao Yuqin, Feng Youji
Department of Gynecology, The Obstetrics and Gynecology Hospital of Fudan University, Shanghai 200011, PR China.
Oncol Rep. 2009 Apr;21(4):893-8. doi: 10.3892/or_00000300.
Our previous study revealed that estrogen regulates nm23-H1 expression thus promoting cell migration-invasion via activating PIK3/Akt pathway. In this study, we explored the effect of hormone on hypoxia-inducible factor-1 (HIF-1alpha), a key factor in cancer invasion and metastasis, via activation of Akt signaling transduction pathway. We treated two ovarian cancer cell lines ES-2 and SKOV3 with 17beta-estradiol, methoxyprogesterone acetate (MPA) only, or hormone combined with and Akt, MAPK pathway inhibitor, or transefected with siRNA targeting Akt sequenced with hormone. Expression of HIF-1alpha was measured by Western blotting. We observed the effect of hormone on nm23-H1 expression after the cells were transfected by siRNA targeting HIF-1alpha or treated with CoCl2 to induce HIF-1alpha overexpression. The 17beta-estradiol increased HIF-1alpha expression in ovarian cancer cells, and this upregulatory effect was abrogated by Akt inhibitor LY294002 (P<0.05) and Akt siRNA interference (P<0.05), but not affected by MAPK inhibitor PD980059 (P>0.05). MPA had the opposite effect. Nm23-H1 protein expression in ES-2 and SKOV3 cells were decreased after treatment with 17beta-estradiol (P<0.05), whereas MPA had the opposite effect. The effect was attenuated by HIF-1alpha siRNA (P<0.05) and enhanced by HIF-1alpha overexpression after CoCl2 treatment (P<0.05). Our data suggest that estrogen and progestin regulate HIF-1alpha expression via Akt signaling pathway, affecting nm23-H1 expression in influencing cell metastasis.
我们之前的研究表明，雌激素通过激活PIK3/Akt通路调节nm23-H1的表达，从而促进细胞迁移侵袭。在本研究中，我们通过激活Akt信号转导通路，探讨了激素对缺氧诱导因子-1（HIF-1α）的影响，HIF-1α是癌症侵袭和转移中的关键因子。我们用17β-雌二醇、醋酸甲羟孕酮（MPA）单独处理，或激素联合Akt、MAPK通路抑制剂处理两种卵巢癌细胞系ES-2和SKOV3，或用靶向Akt的siRNA与激素一起转染细胞。通过蛋白质免疫印迹法检测HIF-1α的表达。在用靶向HIF-1α的siRNA转染细胞或用CoCl2处理诱导HIF-1α过表达后，我们观察了激素对nm23-H1表达的影响。17β-雌二醇增加了卵巢癌细胞中HIF-1α的表达，这种上调作用被Akt抑制剂LY294002（P<0.05）和Akt siRNA干扰（P<0.05）所消除，但不受MAPK抑制剂PD980059的影响（P>0.05）。MPA则有相反的作用。用17β-雌二醇处理后，ES-2和SKOV3细胞中nm23-H1蛋白表达降低（P<0.05），而MPA有相反的作用。该作用被HIF-1α siRNA减弱（P<0.05），并在CoCl2处理后HIF-1α过表达时增强（P<0.05）。我们的数据表明，雌激素和孕激素通过Akt信号通路调节HIF-1α的表达，在影响细胞转移过程中影响nm23-H1的表达。
