鞣花酸結合健擇抑制膀胱癌細胞生長與調解抗藥相關機轉之研究
吳穎羲1、何嘉益3 、于承平2 、高鴻偉2、于大雄4
1國防醫學院病理及寄生蟲研究所，2三軍總醫院病理部，3國防醫學院生命科學研究所，4三軍總醫院泌尿外科
The Study of Growth Inhibition and MDR Modulation of Bladder Cancer by Ellagic Acid Combination with Chemotherapeutics
Ying - Si Wu1, Jar-Yi Ho3, Cheng-Ping Yu2, Hong-Wei Gao2, Dah-Shyong Yu4
1Graduate Institute of Pathology and Parasitology, National Defense Medical Center, 2Department of Pathology, Tri-Service General Hospital, 3Graduate Institute of Life Science, National Defense Medical Center, 4Division of Urology, Department of Surgery, Tri-Service General Hospital
Purpose. Clinically, high incidence and recurrence are accompanied with low survival rates in bladder cancer. Mitomycin C (MMC) and gemcitabine (GCB) are used as common chemotherapeutic agents for bladder cancer patients. About one-third patients shown poor response after MMC or GCB treatment due to drug resistance. Ellagic acid (EA) is a polyphenolic compound that can be extracted from berry plants and is a naturally occurring hydrolysis product. It possesses anti-cancer, anti-angiogenesis, anti-drug resistance, anti-proliferation and apoptosis-inductive activation. Hence, we planned to elucidate whether EA and GCB combined treatment could ameliorate GCB resistance of bladder cancer.
Materials and Methods. High-grade bladder cancer cells T24 and GCB-resistant T24-GCB cells were used. The optimal combined ratios of EA and GCB for treating these bladder cancer cells were evaluated by MTT assays. The effects of combined treatment on cell migration and invasion were evaluated using wound-healing assay and transwell assay and the effects on cell cycle were evaluated by flow cytometry. Proteins regulation of related signal pathway (VEGFR-2, MMP2, MMP9 and EMT process) in bladder cancer cells by EA combined GCB were further evaluated by western blotting.
Result. The most effective drug combination for recurited bladder cancer cells is 30 μM EA combined with 0.1 μM GCB. Using wound healing assays and transwell assays, EA combined with GCB significantly inhibited cell migratory and invasive ability. And using flow cytometry, combined treatment resulted in G1 phase arrest and sub-G1 phase increment in T24-GCB cells. Moreover, protein levels of VEGFR-2, MMP2, MMP9 and EMT-related pathways were down-regulated.
Conclusion. EA combined with low-dose GCB down-regulated the VEGF-related and EMT-related signaling pathways and thereby reduced the drug resistance of T24-GCB cells. These results shad a light to overcome multi-drug resistance (MDR) and assisted chemotherapeutically refractory bladder cancer patients to re-senstize intravesical or systemic chemotherapy.