嘌呤類似物ENERGI-F706藉由活化5'-單磷酸腺苷活化蛋白激酶誘導786-O腎癌細胞的凋亡

許兆畬¹、林君香²、高紹軒²、

童綜合醫院醫院 外科部 泌尿科；²中山醫學大學 醫學院 生化微生物免疫研究所

Purine analogue energi‑f706 induces apoptosis of 786-o renal carcinoma cells via 5'-adenosine monophosphate-activated protein kinase activation

Chao-Yu Hsu¹, Chun-Hsiang Lin², Shao-Hsuan Kao²,

Divisions of Urology, Department of Surgery, Tungs' Taichung MetroHarbor Hospital, Taichung, Taiwan; Institute of Biochemistry, Microbiology and Immunology ², College of Medicine, Chung Shan Medical University, Taichung, Taiwan

Purpose:

Purine compounds are known to activate 5'‑adenosine monophosphate‑activated protein kinase (AMPK), which has important roles in treatments for renal cell carcinoma (RCC). The present study was aimed to investigate the effects of the purine analogue ENERGI‑F706 on the human renal carcinoma cell line 786‑O and the underlying mechanisms.

Materials and Methods:

Cell viability and cell cycle distribution were determined using the MTT assay in the absence or presence of ENERGI‑F706 and flow cytometric analysis, respectively. Phosphorylation and protein levels were assessed by immunoblot analysis. The involvement of AMPK signaling was demonstrated using a specific inhibitor.

Results:

ENERGI‑F706 (0.2‑0.6 mg/ml) significantly decreased the cell viability to up to 36.4±2.4% of that of the control. Compared to 786‑O cells, ENERGI‑F706 exerted less suppressive effects on the viability of the human non‑tumorigenic renal cell line HK‑2. Flow cytometric analysis showed that ENERGI‑F706 contributed to cell cycle arrest at S‑phase and triggered apoptosis of 786‑O cells. Immunoblot analysis revealed that anti‑apoptotic B‑cell lymphoma 2 (Bcl‑2) levels were reduced and pro‑apoptotic Bcl‑2‑associated X protein levels were diminished. In addition, activation of caspase‑9, caspase‑3 and poly(adenosine diphosphate ribose) polymerase (PARP) was promoted in 786‑O cells in response to ENERGI‑F706. Effects of ENERGI‑F706 on AMPK cascades were investi­gated and the results showed that ENERGI‑F706 enhanced phosphorylation of AMPKα (T172) and p53 (S15), a down­stream target of AMPK. In addition, the AMPK activation, p53 (S15) phosphorylation, reduction of Bcl‑2, cleavage of caspase‑3 and PARP as well as suppressed cell viability induced by ENERGI‑F706 were reversed in the presence of AMPK inhibitor compound C (dorsomorphin).