title: Purine analogue ENERGI-F706 induces apoptosis of 786-O renal carcinoma cells via 5'-adenosine monophosphate-activated protein kinase activation.
abstract: Purine compounds are known to activate 5'-adenosine monophosphate-activated protein kinase (AMPK), which has important roles in treatments for renal cell carcinoma. 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. The results revealed that 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). In conclusion, the findings of the present study revealed that ENERGI‑F706 significantly suppressed the viability of 786‑O cells via induction of cell cycle arrest and apoptosis, attributing to AMPK and p53 activation and subsequent cell cycle regulatory and apoptotic signaling. It was therefore indicated that ENERGI‑F706 may be suitable for the treatment of renal cell carcinoma.
authors.affiliation: Division of Urology, Department of Surgery, Tungs' Taichung Metro Harbor Hospital, Taichung 435, Taiwan, R.O.C.
authors.forename: Chao-Yu
authors.initials: CY
authors.lastname: Hsu
authors.affiliation: Institute of Biochemistry and Biotechnology, College of Medicine, Chung Shan Medical University, Taichung 402, Taiwan, R.O.C.
authors.forename: Chun-Hsiang
authors.initials: CH
authors.lastname: Lin
authors.affiliation: Institute of Applied Science and Engineering, Catholic Fu‑Jen University, New Taipei 242, Taiwan, R.O.C.
authors.forename: Jiun-Tsai
authors.initials: JT
authors.lastname: Lin
authors.affiliation: Energenesis Biomedical Co. Ltd., New Taipei 235, Taiwan, R.O.C.
authors.forename: Yi-Fang
authors.initials: YF
authors.lastname: Cheng
authors.affiliation: Institute of Applied Science and Engineering, Catholic Fu‑Jen University, New Taipei 242, Taiwan, R.O.C.
authors.forename: Han-Min
authors.initials: HM
authors.lastname: Chen
authors.affiliation: Institute of Biochemistry and Biotechnology, College of Medicine, Chung Shan Medical University, Taichung 402, Taiwan, R.O.C.
authors.forename: Shao-Hsuan
authors.initials: SH
authors.lastname: Kao
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism; D000972:Antineoplastic Agents, Phytogenic / Q000302:isolation & purification / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D031723:Bambusa / Q000737:chemistry*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D004789:Enzyme Activation / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D009928:Organ Specificity; D010936:Plant Extracts / Q000737:chemistry; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D011687:Purines / Q000302:isolation & purification / Q000494:pharmacology*; D011720:Pyrazoles / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D059807:S Phase Cell Cycle Checkpoints / Q000187:drug effects; D015398:Signal Transduction; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000972:Antineoplastic Agents, Phytogenic; C494826:BAX protein, human; C000595707:BCL2 protein, human; D010936:Plant Extracts; D019253:Proto-Oncogene Proteins c-bcl-2; D011687:Purines; D011720:Pyrazoles; D011743:Pyrimidines; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; C516138:dorsomorphin; D011065:Poly(ADP-ribose) Polymerases; D055372:AMP-Activated Protein Kinases; C526180:PRKAA1 protein, human; D053148:Caspase 3; D053453:Caspase 9