Original Article
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World J Gastroenterol. Nov 28, 2014; 20(44): 16665-16673
Published online Nov 28, 2014. doi: 10.3748/wjg.v20.i44.16665
Estradiol agonists inhibit human LoVo colorectal-cancer cell proliferation and migration through p53
Hsi-Hsien Hsu, Wei-Wen Kuo, Da-Tong Ju, Yu-Lan Yeh, Chuan-Chou Tu, Ying-Lan Tsai, Chia-Yao Shen, Sheng-Huang Chang, Li-Chin Chung, Chih-Yang Huang
Hsi-Hsien Hsu, Division of Colorectal Surgery, Mackay Memorial Hospital, Taipei 251, Taiwan
Hsi-Hsien Hsu, Mackay Junior College of Medicine, Nursing and Management, Taipei 112, Taiwan
Wei-Wen Kuo, Department of Biological Science and Technology, China Medical University, Taichung 404, Taiwan
Da-Tong Ju, Department of Neurological Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan
Yu-Lan Yeh, Department of Pathology, Changhua Christian Hospital, Changhua 500, Taiwan
Chuan-Chou Tu, Division of Chest Medicine, Department of internal Medicine, Armed Force Taichung General Hospital, Taichung 411, Taiwan
Ying-Lan Tsai, National Taiwan Sport University, Athletic Training and Health Department, Taoyuan 333, Taiwan
Chia-Yao Shen, Department of Nursing, MeiHo University, Pingtung 912, Taiwan
Sheng-Huang Chang, Tsao-Tun Psychiatric Center, Department of Health, Executive Yuan, Nantou 542, Taiwan
Li-Chin Chung, Department of Hospital and Health Care Administration, Chia Nan University of Pharmacy and Science, Tainan 717, Taiwan
Chih-Yang Huang, Graduate Institute of Chinese Medical Science, China Medical University, Taichung 404, Taiwan
Chih-Yang Huang, Graduate Institute of Basic Medical Science, China Medical University, Taichung 404, Taiwan
Chih-Yang Huang, Department of Health and Nutrition Biotechnology, Asia University, Taichung 413, Taiwan
Author contributions: All authors contributed to the manuscript.
Supported by Taiwan Department of Health Clinical Trial and Research Center of Excellence No. MOHW103-TDU-B-212-113002
Correspondence to: Chih-Yang Huang, PhD, Graduate Institute of Basic Medical Science, China Medical University, No. 91 Hsueh-Shih Road, Taichung 404, Taiwan. cyhuang@mail.cmu.edu.tw
Telephone: +886-4-22053366-3313 Fax: +886-4-22032295
Received: February 19, 2014
Revised: April 10, 2014
Accepted: June 21, 2014
Published online: November 28, 2014
Processing time: 285 Days and 19.1 Hours
Abstract

AIM: To investigate the effects of 17β-estradiol via estrogen receptors (ER) or direct administration of ER agonists on human colorectal cancer.

METHODS: LoVo cells were established from the Bioresource Collection and Research Center and cultured in phenol red-free DMEM (Sigma, United States). To investigate the effects of E2 and/or ER selective agonists on cellular proliferation, LoVo colorectal cells were treated with E2 or ER-selective agonists for 24 h and 48 h and subjected to the MTT (Sigma) assay to find the concentration. And investigate the effects of E2 and/or ER selective agonists on cell used western immunoblotting to find out the diversification of signaling pathways. In order to observe motility and migration the wound healing assay and a transwell chamber (Neuro Probe) plate were tased. For a quantitative measure, we counted the number of migrating cells to the wound area post-wounding for 24 h. We further examined the cellular migration-regulating factors urokinase-type plasminogen activator (u-PA), tissue-type plasminogen activator (t-PA) and matrix metalloproteinase (MMP)-9 in human LoVo cells so gelatin zymography that we used and gelatinolytic activity was visualized by Coomassie blue staining. And these results are presented as means ± SE, and statistical comparisons were made using Student’s t-test.

RESULTS: The structure was first compared with E2 and ER agonists. We then treated the LoVo cells with E2 and ER agonists (10-8 mol/L) for 24 h and 48 h and subsequently measured the cell viability using MTT assay. Our results showed that treatment with 17β-estradiol and/or ER agonists in human LoVo colorectal cancer cells activated p53 and then up-regulated p21 and p27 protein levels, subsequently inhibiting the downstream target gene, cyclin D1, which regulates cell proliferation. Taken together, our findings demonstrate the anti-tumorigenesis effects of 17β-estradiol and/or ER agonists and suggest that these compounds may prove to be a potential alternative therapy in the treatment of human colorectal cancer. These results demonstrate that 17β-estradiol and/or ER agonists downregulate migration-related proteins through the p53 signaling pathway in human LoVo colorectal cancer cells. These findings suggest that p53 plays a critical role in the 17β-estradiol and/or ER agonist-mediated protective activity against colorectal cancer progression. In addition, 17β-estradiol and/or ER agonists dramatically inhibited cell migration and reduced the expression of u-PA, t-PA and MMP-9 as well as MMP-2/9 activity in LoVo cells, which regulate cell metastasis. Moreover, we observed that pretreatment with a p53 inhibitor significantly blocked the anti-migration effects of E2 and/or ER agonists on LoVo cells. That E2 and/or ER agonists may impair LoVo cell migration by modulating migration-related factors via the p53 tumor suppressor gene.

CONCLUSION: Direct ER treatment may prove to be an attractive alternative therapy in the treatment of human colorectal tumors in the future.

Keywords: Estrogen; Estrogen agonist; Estrogen receptors; Human colon cancer cell; p53

Core tip: The present study is to investigate the effects of 17β-estradiol via estrogen receptors or directly administration of ERs agonist on the development of human colorectal cancer, and to elucidate whether the effect was regulated by tumor suppressor gene p53. Here, our results showed that 17β-estradiol and/or ERs agonist treatment in human LoVo colorectal cancer cells could active p53, then up-regulated p21 and p27 protein levels, subsequently inhibited downstream target gene, cyclin D1, which regulated the cell proliferation.