Liang Gui1,2, Qiang Xu3, Juju Huang4, Gaojue Wu4, Hong Tang4, Li Hui4, Ping Hua4, Lili Zhang4, Yingwei Zhu4. 1. Department of Vascular Surgery, Beijing Hospital, National Center of Gerontology Beijing 100730, China. 2. Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing 100730, China. 3. Department of Intervention, Wujin Hospital Affiliated to Jiangsu University Changzhou 213002, Jiangsu, China. 4. Department of Gastroenterology, The Affiliated Wuxi No. 2 People's Hospital of Nanjing Medical University Wuxi 214002, Jiangsu, China.
Abstract
OBJECTIVE: This study aimed to explore the function and mechanism of Cytochrome P450 2J2 (CYP2J2) epoxygenase and epoxyeicosatrienoic acids (EETs) in the malignant development of hepatocellular carcinoma (HCC). METHOD: The expressional levels of EETs and CYP2J2 in HCC tissues and cell lines were quantified by ELISA, western blot and RT-qPCR, respectively. The effects of EET and CYP2J2 on HCC development were analyzed by CCK-8 assays, flow cytometry analysis, colony formation and transwell assays. The effect of CYP2J2-EET metabolism on stability of HIF-1α was detected by western blot experiments. HIF-1α inhibitor, YC-1, was used to probe the relationship between HIF-1α and metastasis of HCC cells. Finally, xenograft experiments were established to investigate the function of CYP2J2-EETs metabolism in HCC tumorigenesis in vivo. RESULT: CYP2J2, 11, 12-EET and 14, 15-EET were up-regulated in HCC tissues and Huh-7, HepG2 cell lines. Addition of exogenous 14, 15-EET accelerated proliferation and metastasis of HCC cells. Knockdown of CYP2J2 inhibited growth and metastasis of HCC cells and malignant xenograft, which was obviously reversed by addition of 14, 15-EET. Moreover, in Huh-7 and HepG2 cells, CYP2J2-EET metabolism elevated the expression of HIF-1α and its downstream factors including VEGFA, PDK1, GLUT1 and DDIT4 through suppressing the expression of PHD. Treatment of YC-1 remarkably suppressed the HCC cells proliferation and restored the effect of 14, 15-EET on tumor size in vivo. CONCLUSION: The up-regulated levels of CYP2J2 and 14, 15-EET in HCC cells improved the stability of HIF-1α thourgh inhibiting PHD expression, which further promoted the malignant development of HCC. AJTR
OBJECTIVE: This study aimed to explore the function and mechanism of Cytochrome P450 2J2 (CYP2J2) epoxygenase and epoxyeicosatrienoic acids (EETs) in the malignant development of hepatocellular carcinoma (HCC). METHOD: The expressional levels of EETs and CYP2J2 in HCC tissues and cell lines were quantified by ELISA, western blot and RT-qPCR, respectively. The effects of EET and CYP2J2 on HCC development were analyzed by CCK-8 assays, flow cytometry analysis, colony formation and transwell assays. The effect of CYP2J2-EET metabolism on stability of HIF-1α was detected by western blot experiments. HIF-1α inhibitor, YC-1, was used to probe the relationship between HIF-1α and metastasis of HCC cells. Finally, xenograft experiments were established to investigate the function of CYP2J2-EETs metabolism in HCC tumorigenesis in vivo. RESULT: CYP2J2, 11, 12-EET and 14, 15-EET were up-regulated in HCC tissues and Huh-7, HepG2 cell lines. Addition of exogenous 14, 15-EET accelerated proliferation and metastasis of HCC cells. Knockdown of CYP2J2 inhibited growth and metastasis of HCC cells and malignant xenograft, which was obviously reversed by addition of 14, 15-EET. Moreover, in Huh-7 and HepG2 cells, CYP2J2-EET metabolism elevated the expression of HIF-1α and its downstream factors including VEGFA, PDK1, GLUT1 and DDIT4 through suppressing the expression of PHD. Treatment of YC-1 remarkably suppressed the HCC cells proliferation and restored the effect of 14, 15-EET on tumor size in vivo. CONCLUSION: The up-regulated levels of CYP2J2 and 14, 15-EET in HCC cells improved the stability of HIF-1α thourgh inhibiting PHD expression, which further promoted the malignant development of HCC. AJTR
Authors: Yangle Li; Xiaokun Zhao; Huiting Tang; Zhaohui Zhong; Lei Zhang; Ran Xu; Songchao Li; Yi Wang Journal: Urol Int Date: 2011-10-25 Impact factor: 2.089