熱治療抑制膀胱癌細胞經上皮間質轉化所產生之化療抗藥性

陳宏恩1、張安辰2、蔡德甫1、仇光宇1、何肇晏1、黃一勝1,3,4

1新光醫院 泌尿科，2中央研究室；3台北醫學大學 醫學院；4輔仁大學 醫學院

Hyperthermia inhibits chemotherapeutic drug resistance of bladder cancer cells through epithelial mesenchymal transition down-regulation

Hung-En Chen 1, An-Chen Chang2, Te-Fu Tsai 1, Kuang-Yu Chou1, Chao-Yen Ho1 and Thomas I-Sheng Hwang1,3,4

Department of Urology1 and Central Laboratory2, Shin Kong Wu Ho-Su Memorial Hospital; Department of Urology, Taipei Medical University3; Division of Urology, School of Medicine, Fu-Jen Catholic University4, Taipei, Taiwan.

Purpose:

The intravesical chemotherapy has been widely used for bladder cancer (BC), but the development of acquired resistance remains a therapeutic hurdle. In this study, we aim to explore the mechanism of hyperthermia (HT) in inhibiting the action of epithelial cell-mesenchymal transition (EMT)-mediated drug resistance.

Materials and Methods:

Human BC cell line UMUC3 was obtained from Bioresource Collection and Research Center and cultured in a-MEM medium supplemented with 10 % FBS, 2 mM GlutaMAX-1, 100 units/ml penicillin and 100 μg/ml streptomycin maintained at 37 °C under 5 % CO2. BC cells were then treated with HT (43°C) for 24, 48 and 72 hours. Then, Western blot and qPCR were used to analyze the EMT-related proteins and their mRNA expression. Finally, Cell viability was assessed using resazurin reagent to detect whether HT reduces the chemotherapeutic drug resistance of BC cells.

Results:

The Cancer Cell Line Encyclopedia (CCLE), a database contains gene expression, genotype, and drug sensitivity data for human cancer cell lines, was used to screen EMT markers expression in BC cell lines RT4 (grade I), 5637 (grade II) and UMUC3 (grade III). We found cadherin-11 (CDH-11) is a potential candidate participating in EMT-mediated drug resistance. Upon HT stimulation, the UMUC3 cells undergoes morphological changes from mesenchymal to epithelial and demonstrates decreased levels of CDH-11 mRNA and protein expression. Moreover, knockdown CDH-11 expression in UMUC3 cells improve the chemosensitivity of cells to cisplatin, doxorubicin and epirubicin. We also discovered that β-catenin signaling pathway may involved in the function of CDH-11-induced drug resistance. Finally, we established cisplatin-resistance cell line UMUC3cis-R and found that UMUC3cis-R has higher CDH-11 expression compared with wild type UMUC3 cells.

Conclusions:

This study discovered CDH-11/b-catenin axis plays a critical mechanism underlying the development of acquired resistance to chemotherapy and suggest HT or co-targeting CDH-11 as a potential strategy to improve the therapeutic efficacy of intravesical chemotherapy in BC patients.