Cellular effects induced by 17-β-estradiol to reduce the survival of renal cell carcinoma cells.
BACKGROUND
Renal cell carcinoma (RCC) is an adult malignancy with 2:1 men-to-women ratio, which implies the possible role of sex hormones in RCC carcinogenesis. One of the predominant sex hormones in women before menopause, 17-β-estradiol (or E2), may regulate RCC growth by cellular mechanisms that are still not fully understood.


METHODS
The expression levels of E2 receptors (ER1 and ER2) were determined in different RCC cell lines. The DNA damage response induced by E2 was determined by a DNA double-strand break marker γH2AX. To study the possible effect of E2 on oxidative stress response, RCC cells were stained with 2,7-dichlorofluorescein diacetate and analyzed by flow cytometry. Upregulation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2) ser40 phosphorylation in response to oxidative stress was detected by immunoblotting. Finally, annexin V/propidium iodide (PI) double staining assay was used to determine E2-induced cellular apoptosis.


RESULTS
Variable expression of ER1 and ER2 were found in the RCC cell lines studied (786-O, A498, and ACHN), in which ACHN and A498 showed highest and lowest ER expression, respectively. In A498 cells, E2 induced DNA double-strand breaks with positive staining of γH2AX. On the other hand, the level of reactive oxidative species were elevated in ACHN cells after E2 treatment. The E2-induced oxidative stress also induced the Ser40 phosphorylation and nuclear translocation of Nrf2. Finally, we also demonstrated that E2 induced apoptosis as revealed by annexin V/PI double staining.


CONCLUSIONS
In this study, we demonstrated the cellular effects of E2 on DNA repair, ROS production as well as Nrf2 activation, and apoptosis in RCC cell lines. Together these cellular alterations may contribute to the reduced viability of RCC cells following E2 treatment.
Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan.
Sheng-Tang
ST
Wu
School of Medicine, College of Medicine, Fu Jen Catholic University, New Taipei, Taiwan.
Wei-Chi
WC
Ku
Department of Biochemistry, National Defense Medical Center, Taipei, Taiwan.|Department of Medical Research, Cathay General Hospital, Taipei, Taiwan.
Chi-Jung
CJ
Huang
School of Medicine, College of Medicine, Fu Jen Catholic University, New Taipei, Taiwan.|Department of Surgery, Sijhih Cathay General Hospital, No. 2, Ln. 59, Jiancheng Rd., Sijhih Dist., New Taipei, 22174, Taiwan.
Yen-Chieh
YC
Wang
Department of Surgery, Cathay General Hospital, Taipei, Taiwan.
Chih-Ming
CM
Lin
School of Medicine, College of Medicine, Fu Jen Catholic University, New Taipei, Taiwan. skchencmri@gmail.com.|Department of Surgery, Sijhih Cathay General Hospital, No. 2, Ln. 59, Jiancheng Rd., Sijhih Dist., New Taipei, 22174, Taiwan. skchencmri@gmail.com.
Shao-Kuan
SK
Chen
17-β-estradiol; DNA damage; Estrogen receptors; Oxidative stress; Renal cell carcinoma
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