研究組蛋白脫乙醯酶抑制劑阻斷膀胱癌惡性進展與骨轉移機制

吳志緯¹、張安辰²、仇光宇¹、蔡德甫¹、何肇晏¹、黃一勝^1,3,4

¹新光醫院 外科部，²轉譯醫學中心；³台北醫學大學 醫學院；⁴輔仁大學 醫學院

Study of histone deacetylase inhibitors blocking malignant

progression and bone metastasis in human bladder

cancer

Chih-Wei Wu¹, An-Chen Chang², Kuang-Yu Chou¹, Te-Fu Tsai¹, Chao-Yen Ho¹ and Thomas I-Sheng Hwang^1,3,4

Department of Surgery1¹ 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:

Bone metastases from bladder cancer (BC) may cause bone pain, spinal cord compression, and pathological fractures. Compared with other urinary tract malignancies such as prostate cancer or renal cell carcinoma with bone metastases, the prognosis of patients with bone metastases from BC is even worse, showing that skeletal-related events (SRE) caused by BC should not be underestimated. Through literature exploration, it was found that the histone deacetylase inhibitor—Trichostatin A (TSA) inhibits cell proliferation, induces cell apoptosis and promotes cell cycle arrest; However, the functions of TSA on the malignant progression of BC and bone metastasis have not been verified.

Materials and Methods: 

The human BC cells (5637, T24 and UMUC3) were obtained from the Bioresource Collection and Research Center (BCRC; Hsinchu, Taiwan). Resazurin-based cell viability assay and colony formation assay were performed to measure cell proliferation and survival rate, respectively. Western blot was used to measure the levels of indicated protein expression. Cell migrative ability was performed by Wound healing assay. In vitro osteoclast maturation and differentiation were conducted using RAW264.7 cells, which are osteoclast precursors.

Results: 

The resazurin-based cell viability assay showed that TSA did not affect cell survival in the short term. However, colony formation assay demonstrated that TSA significantly inhibited the colony formation of 5637 and UMUC3 BC cells after one week of treatment (long-term). Through a wound healing assay observation for 24 hours, it was found that TSA significantly inhibited the migration ability of BC cells. Moreover, TSA significantly inhibited the anti-anoikis ability of BC cells after 2, 4, and 6 days of treatment. Importantly, we confirmed that BC cells induced osteoclast maturation and differentiation. Mechanismly, TSA inhibited this phenomenon by negatively regulating the NF-κB signaling pathway, demonstrating the potential of TSA in inhibiting osteolytic bone metastasis.

Conclusions:

This study thoroughly examines the mechanisms by which TSA inhibits the malignant progression of BC cells by suppressing their migration, anoikis resistance, and osteoclastogenesis.