紫檀芪對膀胱癌細胞凋亡影響的研究

王啟暐¹、何承勳^{1, 2}、鄭柏青^{3, 4}、劉詩彬⁵

¹新光吳火獅紀念醫院外科部泌尿科

²輔仁大學醫學系

³臺北醫學大學醫學系分子寄生蟲暨熱帶疾病學科

⁴臺北醫學大學國際熱帶醫學研究中心

⁵國立台灣大學醫學院附設醫院泌尿部

The study on the effects of Pterostilbene to cell apoptosis of bladder cancer cells

Chi-Wei Wang¹, Chen-Hsun Ho^{1, 2}, Po-Ching Cheng^{3, 4}, Shih-Ping Liu⁵

¹ Division of Urology, Department of Surgery, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan

² School of Medicine, Fu Jen Catholic University, New Taipei City, Taiwan

³ Department of Molecular Parasitology and Tropical Diseases, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan

⁴ Center for International Tropical Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan

⁵ Department of Urology, National Taiwan University Hospital and College of Medicine, Taipei, Taiwan

Abstract

Introduction: Bladder cancer is the tenth most common cancer in the world, while urothelial carcinoma is the most histological type of bladder cancer, accounting for about 90%. Resecting the tumor or removing the bladder by surgery is the usually clinical treatment. However, this will not only affect the patient's urinary tract system, resulting in poor quality of life, but also cancer cells may metastasize and interfere with postoperative survival. Current research has shifted the focus to bladder-sparing alternative therapies, including apoptosis treatment. Pterostilbene (PT), a natural chemical that is a derivative of resveratrol and has antioxidant, anti-inflammatory and anticancer properties. PT can be used as a candidate drug for inducing apoptosis of cancer cells.

Method: We made an in-depth study on the possible role of PT in regulating the activity of normal bladder cells and tumor cells by analyzing the changes of apoptosis indicators in the process, to precisely explore the effect of PT on bladder cancer. After treating SV-HUC-1 and HTB-9 cells with different concentrations of Pterostilbene for 24 and 48 hours, the cell viability was determined by ELISA and immunofluorescence microscopy. In addition, total cell proteins were also collected for apoptotic protein expression assays.

Results: Our results showed that in cell viability assays, increasing PT concentrations resulted in a significant decrease in the viability of HTB-9 cells (>10uM), compared to normal SV-HUC-1 cells which requiring higher concentrations for a significant decrease (>50uM). The expression of specific protein indicators further showed that the cell activity inhibited by PT was related to the expression of NF-kB, BcL-2, Bax and other apoptotic proteins. Moreover, the Annexin-V/PI staining of PT-treated SV-HUC-1 and HTB-9 cells was observed by fluorescence microscope; and found that with the increase of PT concentration, cell death with PI-staining also increased, especially in the 48-hours treated group of HTB-9 cancer cell line. Meanwhile, Caspase 3 fluorescent staining detection also showed similar results.

Conclusion: Our results found that PT could induce apoptosis and inhibit the growth of HTB-9 bladder cancer cells, and bladder cancer cells were more sensitive to the apoptotic effect of PT than normal bladder cell lines. Although PT induces significantly more apoptosis in HTB-9 cells than SV-HUC-1 cells, more data are needed to elucidate the mechanism by which PT promotes apoptosis of bladder cancer cells. PT combining with current bladder cancer drugs to be developed as additional adjuvant treatment are directions worth exploring.

Keyword: Bladder cancer; Pterostilbene; apoptosis; SV-HUC-1 cells; HTB-9 cells