Targeted inhibition of mitochondrial Hsp90 induces mitochondrial elongation in Hep3B hepatocellular carcinoma cells undergoing apoptosis by increasing the ROS level

Yoo,Seung Hee; Kim,Hye  Young; Rho,Jee  Hyun; Jeong,Seon-Yong; Yun,Jeanho; Yun,Il; Park,Hwan  Tae; Yoo,Young  Hyun

doi:10.3892/ijo.2015.3150

Targeted inhibition of mitochondrial Hsp90 induces mitochondrial elongation in Hep3B hepatocellular carcinoma cells undergoing apoptosis by increasing the ROS level

Authors:
- Seung Hee Yoo
- Hye Young Kim
- Jee Hyun Rho
- Seon-Yong Jeong
- Jeanho Yun
- Il Yun
- Hwan Tae Park
- Young Hyun Yoo
View Affiliations

Affiliations: Department of Anatomy and Cell Biology and Mitochondria Hub Regulation Center, College of Medicine, Dong-A University, Busan, Republic of Korea, Department of Medical Genetics, School of Medicine, Ajou University, Suwon, Republic of Korea, Department of Biochemistry, Dong-A University College of Medicine, Busan, Republic of Korea, Department of Dental Pharmacology and Biophysics, School of Dentistry and Research Institute for Oral Biotechnology, Yangsan Campus of Pusan National University, Yangsan, Republic of Korea, Department of Physiology and Mitochondria Hub Regulation Center, Dong-A University College of Medicine, Busan, Republic of Korea
Published online on: September 7, 2015 https://doi.org/10.3892/ijo.2015.3150
Pages: 1783-1792

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Previous studies reported that a Gamitrinib variant containing triphenylphosphonium (G-TPP) binds to mitochondrial Hsp90 and rapidly inhibits its activity to induce apoptosis. We investigated the mechanisms underlying the antitumor activity of G-TPP in Hep3B hepatocellular carcinoma cells. Contrary to our predictions, we observed mitochondrial elongation in the G-TPP-treated Hep3B cells undergoing apoptosis. We found that the G-TPP-induced mitochondrial elongation in Hep3B cells was caused by a decrease in the mitochondrial fission-regulating protein Drp1 rather than by changes in the mitochondrial fusion machinery proteins Mfn1 and Opa1. Furthermore, G-TPP induced G2-M phase cell cycle arrest by reducing the interaction between CDK1 and cyclin B1. Additionally, reactive oxygen species (ROS) played a pivotal role in G-TPP-induced cell death and mitochondrial elongation in Hep3B cells, and these processes are mediated by the reduced association of CDK1 with cyclin B1 and the suppressed phosphorylation of Drp1 (Ser616). Thus, G-TPP induces cell death and causes Drp1-mediated mitochondrial elongation in Hep3B cells by increasing the ROS level.

View Figures

View References

1	Campello S, Strappazzon F and Cecconi F: Mitochondrial dismissal in mammals, from protein degradation to mitophagy. Biochim Biophys Acta. 1837:451–460. 2014. View Article : Google Scholar
2	Picard M, Shirihai OS, Gentil BJ and Burelle Y: Mitochondrial morphology transitions and functions: Implications for retrograde signaling? Am J Physiol Regul Integr Comp Physiol. 304:R393–R406. 2013. View Article : Google Scholar : PubMed/NCBI
3	Harbauer AB, Zahedi RP, Sickmann A, Pfanner N and Meisinger C: The protein import machinery of mitochondria - a regulatory hub in metabolism, stress, and disease. Cell Metab. 19:357–372. 2014. View Article : Google Scholar : PubMed/NCBI
4	Karbowski M and Youle RJ: Dynamics of mitochondrial morphology in healthy cells and during apoptosis. Cell Death Differ. 10:870–880. 2003. View Article : Google Scholar : PubMed/NCBI
5	Cerveny KL, Tamura Y, Zhang Z, Jensen RE and Sesaki H: Regulation of mitochondrial fusion and division. Trends Cell Biol. 17:563–569. 2007. View Article : Google Scholar : PubMed/NCBI
6	Benard G and Karbowski M: Mitochondrial fusion and division: Regulation and role in cell viability. Semin Cell Dev Biol. 20:365–374. 2009. View Article : Google Scholar : PubMed/NCBI
7	Ishihara N, Fujita Y, Oka T and Mihara K: Regulation of mitochondrial morphology through proteolytic cleavage of OPA1. EMBO J. 25:2966–2977. 2006. View Article : Google Scholar : PubMed/NCBI
8	Smirnova E, Griparic L, Shurland DL and van der Bliek AM: Dynamin-related protein Drp1 is required for mitochondrial division in mammalian cells. Mol Biol Cell. 12:2245–2256. 2001. View Article : Google Scholar : PubMed/NCBI
9	Landes T and Martinou JC: Mitochondrial outer membrane permeabilization during apoptosis: The role of mitochondrial fission. Biochim Biophys Acta. 1813:540–545. 2011. View Article : Google Scholar : PubMed/NCBI
10	Otera H, Wang C, Cleland MM, Setoguchi K, Yokota S, Youle RJ and Mihara K: Mff is an essential factor for mitochondrial recruitment of Drp1 during mitochondrial fission in mammalian cells. J Cell Biol. 191:1141–1158. 2010. View Article : Google Scholar : PubMed/NCBI
11	Twig G, Elorza A, Molina AJ, Mohamed H, Wikstrom JD, Walzer G, Stiles L, Haigh SE, Katz S, Las G, et al: Fission and selective fusion govern mitochondrial segregation and elimination by autophagy. EMBO J. 27:433–446. 2008. View Article : Google Scholar : PubMed/NCBI
12	Eisenberg-Lerner A, Bialik S, Simon HU and Kimchi A: Life and death partners: Apoptosis, autophagy and the cross-talk between them. Cell Death Differ. 16:966–975. 2009. View Article : Google Scholar : PubMed/NCBI
13	Wang IH, Chen HY, Wang YH, Chang KW, Chen YC and Chang CR: Resveratrol modulates mitochondria dynamics in replicative senescent yeast cells. PLoS One. 9:e1043452014. View Article : Google Scholar : PubMed/NCBI
14	Pinton P, Ferrari D, Rapizzi E, Di Virgilio F, Pozzan T and Rizzuto R: The Ca²⁺ concentration of the endoplasmic reticulum is a key determinant of ceramide-induced apoptosis: Significance for the molecular mechanism of Bcl-2 action. EMBO J. 20:2690–2701. 2001. View Article : Google Scholar : PubMed/NCBI
15	Breckenridge DG, Stojanovic M, Marcellus RC and Shore GC: Caspase cleavage product of BAP31 induces mitochondrial fission through endoplasmic reticulum calcium signals, enhancing cytochrome c release to the cytosol. J Cell Biol. 160:1115–1127. 2003. View Article : Google Scholar : PubMed/NCBI
16	Bai X, Yan Y, Canfield S, Muravyeva MY, Kikuchi C, Zaja I, Corbett JA and Bosnjak ZJ: Ketamine enhances human neural stem cell proliferation and induces neuronal apoptosis via reactive oxygen species-mediated mitochondrial pathway. Anesth Analg. 116:869–880. 2013. View Article : Google Scholar : PubMed/NCBI
17	Atay C, Ugurlu S and Ozören N: Shock the heat shock network. J Clin Invest. 119:445–448. 2009. View Article : Google Scholar : PubMed/NCBI
18	Kang BH, Plescia J, Song HY, Meli M, Colombo G, Beebe K, Scroggins B, Neckers L and Altieri DC: Combinatorial drug design targeting multiple cancer signaling networks controlled by mitochondrial Hsp90. J Clin Invest. 119:454–464. 2009. View Article : Google Scholar : PubMed/NCBI
19	Mayer MP, Prodromou C and Frydman J: The Hsp90 mosaic: A picture emerges. Nat Struct Mol Biol. 16:2–6. 2009. View Article : Google Scholar : PubMed/NCBI
20	Yamano K and Youle RJ: Coupling mitochondrial and cell division. Nat Cell Biol. 13:1026–1027. 2011. View Article : Google Scholar : PubMed/NCBI
21	Park HK, Lee JE, Lim J and Kang BH: Mitochondrial Hsp90s suppress calcium-mediated stress signals propagating from mitochondria to the ER in cancer cells. Mol Cancer. 13:1482014. View Article : Google Scholar : PubMed/NCBI
22	Siegelin MD, Dohi T, Raskett CM, Orlowski GM, Powers CM, Gilbert CA, Ross AH, Plescia J and Altieri DC: Exploiting the mitochondrial unfolded protein response for cancer therapy in mice and human cells. J Clin Invest. 121:1349–1360. 2011. View Article : Google Scholar : PubMed/NCBI
23	Frank S, Gaume B, Bergmann-Leitner ES, Leitner WW, Robert EG, Catez F, Smith CL and Youle RJ: The role of dynamin-related protein 1, a mediator of mitochondrial fission, in apoptosis. Dev Cell. 1:515–525. 2001. View Article : Google Scholar : PubMed/NCBI
24	Lee JS, Yoon YG, Yoo SH, Jeong NY, Jeong SH, Lee SY, Jung DI, Jeong SY and Yoo YH: Histone deacetylase inhibitors induce mitochondrial elongation. J Cell Physiol. 227:2856–2869. 2012. View Article : Google Scholar
25	Abeliovich H: Mitophagy: The life-or-death dichotomy includes yeast. Autophagy. 3:275–277. 2007. View Article : Google Scholar : PubMed/NCBI
26	Narendra D, Tanaka A, Suen DF and Youle RJ: Parkin is recruited selectively to impaired mitochondria and promotes their autophagy. J Cell Biol. 183:795–803. 2008. View Article : Google Scholar : PubMed/NCBI
27	Twig G and Shirihai OS: The interplay between mitochondrial dynamics and mitophagy. Antioxid Redox Signal. 14:1939–1951. 2011. View Article : Google Scholar :
28	Ni HM, Williams JA and Ding WX: Mitochondrial dynamics and mitochondrial quality control. Redox Biol. 4:6–13. 2015. View Article : Google Scholar :
29	Arnoult D, Rismanchi N, Grodet A, Roberts RG, Seeburg DP, Estaquier J, Sheng M and Blackstone C: Bax/Bak-dependent release of DDP/TIMM8a promotes Drp1-mediated mitochondrial fission and mitoptosis during programmed cell death. Curr Biol. 15:2112–2118. 2005. View Article : Google Scholar : PubMed/NCBI
30	Bernhardt D, Müller M, Reichert AS and Osiewacz HD: Simultaneous impairment of mitochondrial fission and fusion reduces mitophagy and shortens replicative lifespan. Sci Rep. 5:78852015. View Article : Google Scholar : PubMed/NCBI
31	Deng H, Dodson MW, Huang H and Guo M: The Parkinson's disease genes pink1 and parkin promote mitochondrial fission and/or inhibit fusion in Drosophila. Proc Natl Acad Sci USA. 105:14503–14508. 2008. View Article : Google Scholar : PubMed/NCBI
32	Poole AC, Thomas RE, Andrews LA, McBride HM, Whitworth AJ and Pallanck LJ: The PINK1/Parkin pathway regulates mitochondrial morphology. Proc Natl Acad Sci USA. 105:1638–1643. 2008. View Article : Google Scholar : PubMed/NCBI
33	Kageyama Y, Hoshijima M, Seo K, Bedja D, Sysa-Shah P, Andrabi SA, Chen W, Höke A, Dawson VL, Dawson TM, et al: Parkin-independent mitophagy requires Drp1 and maintains the integrity of mammalian heart and brain. EMBO J. 33:2798–2813. 2014. View Article : Google Scholar : PubMed/NCBI
34	Hirota Y, Kang D and Kanki T: The physiological role of mitophagy: New insights into phosphorylation events. Int J Cell Biol. 2012:3549142012. View Article : Google Scholar : PubMed/NCBI
35	Mitra K, Wunder C, Roysam B, Lin G and Lippincott-Schwartz J: A hyperfused mitochondrial state achieved at G1-S regulates cyclin E buildup and entry into S phase. Proc Natl Acad Sci USA. 106:11960–11965. 2009. View Article : Google Scholar : PubMed/NCBI
36	Qian W, Choi S, Gibson GA, Watkins SC, Bakkenist CJ and Van Houten B: Mitochondrial hyperfusion induced by loss of the fission protein Drp1 causes ATM-dependent G2/M arrest and aneuploidy through DNA replication stress. J Cell Sci. 125:5745–5757. 2012. View Article : Google Scholar : PubMed/NCBI
37	Westrate LM, Sayfie AD, Burgenske DM and MacKeigan JP: Persistent mitochondrial hyperfusion promotes G2/M accumulation and caspase-dependent cell death. PLoS One. 9:e919112014. View Article : Google Scholar : PubMed/NCBI
38	Huang WW, Ko SW, Tsai HY, Chung JG, Chiang JH, Chen KT, Chen YC, Chen HY, Chen YF and Yang JS: Cantharidin induces G2/M phase arrest and apoptosis in human colorectal cancer colo 205 cells through inhibition of CDK1 activity and caspase-dependent signaling pathways. Int J Oncol. 38:1067–1073. 2011.PubMed/NCBI
39	Cribbs JT and Strack S: Reversible phosphorylation of Drp1 by cyclic AMP-dependent protein kinase and calcineurin regulates mitochondrial fission and cell death. EMBO Rep. 8:939–944. 2007. View Article : Google Scholar : PubMed/NCBI
40	Chang CR and Blackstone C: Cyclic AMP-dependent protein kinase phosphorylation of Drp1 regulates its GTPase activity and mitochondrial morphology. J Biol Chem. 282:21583–21587. 2007. View Article : Google Scholar : PubMed/NCBI
41	Taguchi N, Ishihara N, Jofuku A, Oka T and Mihara K: Mitotic phosphorylation of dynamin-related GTPase Drp1 participates in mitochondrial fission. J Biol Chem. 282:11521–11529. 2007. View Article : Google Scholar : PubMed/NCBI
42	Yang Y, Bazhin AV, Werner J and Karakhanova S: Reactive oxygen species in the immune system. Int Rev Immunol. 32:249–270. 2013. View Article : Google Scholar : PubMed/NCBI
43	Matsuzawa A and Ichijo H: Redox control of cell fate by MAP kinase: Physiological roles of ASK1-MAP kinase pathway in stress signaling. Biochim Biophys Acta. 1780:1325–1336. 2008. View Article : Google Scholar : PubMed/NCBI
44	Zhang Y, Du Y, Le W, Wang K, Kieffer N and Zhang J: Redox control of the survival of healthy and diseased cells. Antioxid Redox Signal. 15:2867–2908. 2011. View Article : Google Scholar : PubMed/NCBI
45	Chandel NS: Mitochondria as signaling organelles. BMC Biol. 12:342014. View Article : Google Scholar : PubMed/NCBI
46	Yu T, Robotham JL and Yoon Y: Increased production of reactive oxygen species in hyperglycemic conditions requires dynamic change of mitochondrial morphology. Proc Natl Acad Sci USA. 103:2653–2658. 2006. View Article : Google Scholar : PubMed/NCBI
47	Yu T, Sheu SS, Robotham JL and Yoon Y: Mitochondrial fission mediates high glucose-induced cell death through elevated production of reactive oxygen species. Cardiovasc Res. 79:341–351. 2008. View Article : Google Scholar : PubMed/NCBI
48	Kageyama Y, Zhang Z, Roda R, Fukaya M, Wakabayashi J, Wakabayashi N, Kensler TW, Reddy PH, Iijima M and Sesaki H: Mitochondrial division ensures the survival of postmitotic neurons by suppressing oxidative damage. J Cell Biol. 197:535–551. 2012. View Article : Google Scholar : PubMed/NCBI
49	Chiche J, Rouleau M, Gounon P, Brahimi-Horn MC, Pouysségur J and Mazure NM: Hypoxic enlarged mitochondria protect cancer cells from apoptotic stimuli. J Cell Physiol. 222:648–657. 2010.
50	Yoon YS, Yoon DS, Lim IK, Yoon SH, Chung HY, Rojo M, Malka F, Jou MJ, Martinou JC and Yoon G: Formation of elongated giant mitochondria in DFO-induced cellular senescence: Involvement of enhanced fusion process through modulation of Fis1. J Cell Physiol. 209:468–480. 2006. View Article : Google Scholar : PubMed/NCBI