Trichostatin A, a histone deacetylase inhibitor, induces synergistic cytotoxicity with chemotherapy via suppression of Raf/MEK/ERK pathway in urothelial carcinoma
- 473 Downloads
In this study, we aimed to investigate the antitumor effects of trichostatin A (TSA), an antifungal antibiotic that inhibits histone deacetylase (HDAC) family of enzymes, alone or in combination with anyone of the three chemotherapeutic agents (cisplatin, gemcitabine, and doxorubicin) for the treatment of human urothelial carcinoma (UC). Two high-grade human UC cell lines (T24 and NTUB1) were used. Cytotoxicity and apoptosis were assessed by MTT assay and flow cytometry, respectively. The expression of phospho-c-Raf, phospho-MEK1/2, and phospho-ERK1/2 was measured by western blotting. ERK siRNA knockdown and the specific MEK inhibitor U0126 were used to examine the role of Raf/MEK/ERK signaling pathway in combined cytotoxicity of TSA and chemotherapy. TSA co-treatment with any one of the three chemotherapeutic agents induced synergistic cytotoxicity (combination index < 1) and concomitantly suppressed chemotherapeutic drug-induced activation of Raf-MEK-ERK pathway. Combination of ERK siRNA knockdown and treatment with the specific MEK inhibitor (U0126) enhanced the cytotoxic effects of the chemotherapy on UC cells. These observations were confirmed in a xenograft nude mouse model. Moreover, activated Raf/MEK/ERK pathway was observed in human bladder UC specimens from patients with chemoresistant status. In conclusion, TSA elicits a synergistic cytotoxic response in combination with chemotherapy via targeting the Raf/MEK/ERK pathway. TSA elicits synergistic cytotoxic response in combination with three DNA-damaging drugs (cisplatin, gemcitabine, and doxorubicin). Activated Raf/MEK/ERK pathway is involved in chemoresistant mechanism of UC. Combining chemotherapeutic agents with HDAC inhibitor (TSA) or with targeting Raf/MEK/ERK pathway is promising to circumvent chemoresistance in UCs.
KeywordsUrothelial carcinoma Trichostatin A Histone deacetylase inhibitor Chemotherapy Drug resistance
extracellular signal-regulated kinase 1 and 2 (ERK1/2)
This work was supported by grants from the Ministry of Science and Technology of Taiwan (104-2314-B-002-164-MY3 and 103-2314-B-002-161-MY3), National Taiwan University Hospital (103-S2349, 104-M2868, 105-S2978,105-28, 106-3415, and 107-S3784), and New Taipei City Hospital.
Wei-Chou Lin, Kuan-Lin Kuo, and Kuo-How Huang conceived of the presented idea and study design.
Kuan-Lin Kuo and Kuo-How Huang wrote the manuscript with support from Fu-Shun Hsu and Wei-Chou Lin.
Kuan-Lin Kuo, Shih-Ming Liao, Jo-Yu Hong, and Shao-Ping Yang carried out the experiment and performed the computations.
Kuo-How Huang, Shing-Hwa Liu, Chung-Sheng Shi, Hong-Chiang Chang, Fu-Shun Hsu, Wei-Chou Lin, Yu-Chieh Tsai, and June-Tai Wu verified the analytical methods and helped supervise the project.
Wei-Chou Lin and Chia-Dong Shun interpreted the results of immunostaining.
All authors discussed the results and contributed to the final manuscript.
We also thank the personnel of the Second, Third, and Sixth Core Laboratories of National Taiwan University Hospital.
Compliance with ethical standards
The study that involve human participants and animal experiments have been approved by the institutional research ethics committee (no. 201112136RIC) and National Taiwan University College of Medicine and College of Public Health Institutional Animal Care and Use Committee (IACUC) (No. 20160117).
Conflict of interest
The authors declare that they have no conflict of interest.
- 2.Harker WG, Meyers FJ, Freiha FS, Palmer JM, Shortliffe LD, Hannigan JF, McWhirter KM, Torti FM (1985) Cisplatin, methotrexate, and vinblastine (CMV): an effective chemotherapy regimen for metastatic transitional cell carcinoma of the urinary tract. A northern California oncology group study. J Clin Oncol 3:1463–1470CrossRefGoogle Scholar
- 3.Sternberg CN, Yagoda A, Scher HI, Watson RC, Geller N, Herr HW, Morse MJ, Sogani PC, Vaughan ED, Bander N, Weiselberg L, Rosado K, Smart T, Lin SY, Penenberg D, Fair WR, Whitmore WF (1989) Methotrexate, vinblastine, doxorubicin, and cisplatin for advanced transitional cell carcinoma of the urothelium. Efficacy and patterns of response and relapse. Cancer 64:2448–2458CrossRefGoogle Scholar
- 4.von der MH HSW, Roberts JT, Dogliotti L, Oliver T, Moore MJ, Bodrogi I, Albers P, Knuth A, Lippert CM et al (2000) Gemcitabine and cisplatin versus methotrexate, vinblastine, doxorubicin, and cisplatin in advanced or metastatic bladder cancer: results of a large, randomized, multinational, multicenter, phase III study. J Clin Oncol 18:3068–3077CrossRefGoogle Scholar
- 21.Yoon CY, Park MJ, Lee JS, Lee SC, Oh JJ, Park H, Chung CW, Abdullajanov MM, Jeong SJ, Hong SK, Byun SS, Lee ES, Lee SE (2011) The histone deacetylase inhibitor trichostatin a synergistically resensitizes a cisplatin resistant human bladder cancer cell line. J Urol 185:1102–1111CrossRefGoogle Scholar