Targeting bortezomib-induced aggresome formation using vinorelbine enhances the cytotoxic effect along with ER stress loading in breast cancer cell lines

Miyahara,Kana; Kazama,Hiromi; Kokuba,Hiroko; Komatsu,Seiichiro; Hirota,Ayako; Takemura,Jun; Hirasawa,Kazuhiro; Moriya,Shota; Abe,Akihisa; Hiramoto,Masaki; Ishikawa,Takashi; Miyazawa,Keisuke

doi:10.3892/ijo.2016.3673

Targeting bortezomib-induced aggresome formation using vinorelbine enhances the cytotoxic effect along with ER stress loading in breast cancer cell lines

Authors:
- Kana Miyahara
- Hiromi Kazama
- Hiroko Kokuba
- Seiichiro Komatsu
- Ayako Hirota
- Jun Takemura
- Kazuhiro Hirasawa
- Shota Moriya
- Akihisa Abe
- Masaki Hiramoto
- Takashi Ishikawa
- Keisuke Miyazawa
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Affiliations: Department of Breast Surgery, Tokyo Medical University, Tokyo, Japan, Department of Biochemistry, Tokyo Medical University, Tokyo, Japan, Laboratory of Electron Microscopy, Tokyo Medical University, Tokyo, Japan, Department of Otolaryngology (Head and Neck Surgery), Tokyo Medical University, Tokyo, Japan
Published online on: August 29, 2016 https://doi.org/10.3892/ijo.2016.3673
Pages: 1848-1858
Copyright: © Miyahara et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The ubiquitin-proteasome and autophagy-lysosome pathways are two major self-digestive systems for cellular proteins. Ubiquitinated misfolded proteins are degraded mostly by proteasome. However, when ubiquitinated proteins accumulate beyond the capacity of proteasome clearance, they are transported to the microtubule-organizing center (MTOC) along the microtubules to form aggresomes, and subsequently some of them are degraded by the autophagy-lysosome system. We previously reported that macrolide antibiotics such as azithromycin and clarithromycin block autophagy flux, and that concomitant treatment with the proteasome inhibitor bortezomib (BZ) and macrolide enhances endoplasmic reticulum (ER) stress-mediated apoptosis in breast cancer cells. As ubiquitinated proteins are concentrated at the aggresome upon proteasome failure, we focused on the microtubule as the scaffold of this transport pathway for aggresome formation. Treatment of metastatic breast cancer cell lines (e.g., MDA-MB‑231 cells) with BZ resulted in induction of aggresomes, which immunocytochemistry detected as a distinctive eyeball-shaped vimentin-positive inclusion body that formed in a perinuclear lesion, and that electron microscopy detected as a sphere of fibrous structure with some dense amorphous deposit. Vinorelbine (VNR), which inhibits microtubule polymerization, more effectively suppressed BZ-induced aggresome formation than paclitaxel (PTX), which stabilizes microtubules. Combined treatment using BZ and VNR, but not PTX, enhanced the cytotoxic effect and apoptosis induction along with pronounced ER stress loading such as upregulation of GRP78 and CHOP/GADD153. The addition of azithromycin to block autophagy flux in the BZ plus VNR-containing cell culture further enhanced the cytotoxicity. These data suggest that suppression of BZ-induced aggresome formation using an inhibitory drug such as VNR for microtubule polymerization is a novel strategy for metastatic breast cancer therapy.

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1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Runowicz CD, Leach CR, Henry NL, Henry KS, Mackey HT, Cowens-Alvarado RL, Cannady RS, Pratt-Chapman ML, Edge SB, Jacobs LA, et al; American Cancer Society/American Society of Clinical Oncology. Breast Cancer Survivorship Care Guideline. J Clin Oncol. 34:611–635. 2016. View Article : Google Scholar
3	Veldhoen RA, Banman SL, Hemmerling DR, Odsen R, Simmen T, Simmonds AJ, Underhill DA and Goping IS: The chemotherapeutic agent paclitaxel inhibits autophagy through two distinct mechanisms that regulate apoptosis. Oncogene. 32:736–746. 2013. View Article : Google Scholar
4	Wen J, Yeo S, Wang C, Chen S, Sun S, Haas MA, Tu W, Jin F and Guan JL: Autophagy inhibition re-sensitizes pulse stimulation-selected paclitaxel-resistant triple negative breast cancer cells to chemotherapy-induced apoptosis. Breast Cancer Res Treat. 149:619–629. 2015. View Article : Google Scholar : PubMed/NCBI
5	Liu S and Li X: Autophagy inhibition enhances sensitivity of endometrial carcinoma cells to paclitaxel. Int J Oncol. 46:2399–2408. 2015.PubMed/NCBI
6	Wojcik S: Crosstalk between autophagy and proteasome protein degradation systems: Possible implications for cancer therapy. Folia Histochem Cytobiol. 51:249–264. 2013. View Article : Google Scholar
7	Mizushima N: Autophagy: Process and function. Genes Dev. 21:2861–2873. 2007. View Article : Google Scholar : PubMed/NCBI
8	Levine B and Kroemer G: Autophagy in the pathogenesis of disease. Cell. 132:27–42. 2008. View Article : Google Scholar : PubMed/NCBI
9	Korolchuk VI, Menzies FM and Rubinsztein DC: Mechanisms of cross-talk between the ubiquitin-proteasome and autophagy-lysosome systems. FEBS Lett. 584:1393–1398. 2010. View Article : Google Scholar
10	Kirkin V, McEwan DG, Novak I and Dikic I: A role for ubiquitin in selective autophagy. Mol Cell. 34:259–269. 2009. View Article : Google Scholar : PubMed/NCBI
11	Degenhardt K, Mathew R, Beaudoin B, Bray K, Anderson D, Chen G, Mukherjee C, Shi Y, Gélinas C, Fan Y, et al: Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis. Cancer Cell. 10:51–64. 2006. View Article : Google Scholar : PubMed/NCBI
12	Mathew R, Karp CM, Beaudoin B, Vuong N, Chen G, Chen HY, Bray K, Reddy A, Bhanot G, Gelinas C, et al: Autophagy suppresses tumorigenesis through elimination of p62. Cell. 137:1062–1075. 2009. View Article : Google Scholar : PubMed/NCBI
13	Karantza-Wadsworth V, Patel S, Kravchuk O, Chen G, Mathew R, Jin S and White E: Autophagy mitigates metabolic stress and genome damage in mammary tumorigenesis. Genes Dev. 21:1621–1635. 2007. View Article : Google Scholar : PubMed/NCBI
14	Guo JY, Chen HY, Mathew R, Fan J, Strohecker AM, Karsli-Uzunbas G, Kamphorst JJ, Chen G, Lemons JM, Karantza V, et al: Activated Ras requires autophagy to maintain oxidative metabolism and tumorigenesis. Genes Dev. 25:460–470. 2011. View Article : Google Scholar : PubMed/NCBI
15	Kawaguchi T, Miyazawa K, Moriya S, Ohtomo T, Che XF, Naito M, Itoh M and Tomoda A: Combined treatment with bortezomib plus bafilomycin A1 enhances the cytocidal effect and induces endoplasmic reticulum stress in U266 myeloma cells: Crosstalk among proteasome, autophagy-lysosome and ER stress. Int J Oncol. 38:643–654. 2011.
16	Komatsu S, Miyazawa K, Moriya S, Takase A, Naito M, Inazu M, Kohno N, Itoh M and Tomoda A: Clarithromycin enhances bortezomib-induced cytotoxicity via endoplasmic reticulum stress-mediated CHOP (GADD153) induction and autophagy in breast cancer cells. Int J Oncol. 40:1029–1039. 2012.
17	Moriya S, Che XF, Komatsu S, Abe A, Kawaguchi T, Gotoh A, Inazu M, Tomoda A and Miyazawa K: Macrolide antibiotics block autophagy flux and sensitize to bortezomib via endoplasmic reticulum stress-mediated CHOP induction in myeloma cells. Int J Oncol. 42:1541–1550. 2013.PubMed/NCBI
18	Meusser B, Hirsch C, Jarosch E and Sommer T: ERAD: The long road to destruction. Nat Cell Biol. 7:766–772. 2005. View Article : Google Scholar : PubMed/NCBI
19	Hetz C: The unfolded protein response: Controlling cell fate decisions under ER stress and beyond. Nat Rev Mol Cell Biol. 13:89–102. 2012.PubMed/NCBI
20	Smith MH, Ploegh HL and Weissman JS: Road to ruin: Targeting proteins for degradation in the endoplasmic reticulum. Science. 334:1086–1090. 2011. View Article : Google Scholar : PubMed/NCBI
21	Brodsky JL: Cleaning up: ER-associated degradation to the rescue. Cell. 151:1163–1167. 2012. View Article : Google Scholar : PubMed/NCBI
22	Sano R and Reed JC: ER stress-induced cell death mechanisms. Biochim Biophys Acta. 1833.3460–3470. 2013.
23	Oyadomari S and Mori M: Roles of CHOP/GADD153 in endoplasmic reticulum stress. Cell Death Differ. 11:381–389. 2004. View Article : Google Scholar
24	Tabas I and Ron D: Integrating the mechanisms of apoptosis induced by endoplasmic reticulum stress. Nat Cell Biol. 13:184–190. 2011. View Article : Google Scholar : PubMed/NCBI
25	Verfaillie T, Garg AD and Agostinis P: Targeting ER stress induced apoptosis and inflammation in cancer. Cancer Lett. 332:249–264. 2013. View Article : Google Scholar
26	Li J, Ni M, Lee B, Barron E, Hinton DR and Lee AS: The unfolded protein response regulator GRP78/BiP is required for endoplasmic reticulum integrity and stress-induced autophagy in mammalian cells. Cell Death Differ. 15:1460–1471. 2008. View Article : Google Scholar : PubMed/NCBI
27	Johnston JA, Ward CL and Kopito RR: Aggresomes: A cellular response to misfolded proteins. J Cell Biol. 143:1883–1898. 1998. View Article : Google Scholar : PubMed/NCBI
28	Kawaguchi Y, Kovacs JJ, McLaurin A, Vance JM, Ito A and Yao TP: The deacetylase HDAC6 regulates aggresome formation and cell viability in response to misfolded protein stress. Cell. 115:727–738. 2003. View Article : Google Scholar : PubMed/NCBI
29	Olzmann JA and Chin LS: Parkin-mediated K63-linked polyubiquitination: A signal for targeting misfolded proteins to the aggresome-autophagy pathway. Autophagy. 4:85–87. 2008. View Article : Google Scholar
30	Lee JY, Koga H, Kawaguchi Y, Tang W, Wong E, Gao YS, Pandey UB, Kaushik S, Tresse E, Lu J, et al: HDAC6 controls autophagosome maturation essential for ubiquitin-selective quality-control autophagy. EMBO J. 29:969–980. 2010. View Article : Google Scholar : PubMed/NCBI
31	Rodriguez-Gonzalez A, Lin T, Ikeda AK, Simms-Waldrip T, Fu C and Sakamoto KM: Role of the aggresome pathway in cancer: Targeting histone deacetylase 6-dependent protein degradation. Cancer Res. 68:2557–2560. 2008. View Article : Google Scholar : PubMed/NCBI
32	Kopito RR: Aggresomes, inclusion bodies and protein aggregation. Trends Cell Biol. 10:524–530. 2000. View Article : Google Scholar : PubMed/NCBI
33	Brüning A and Jückstock J: Misfolded proteins: From little villains to little helpers in the fight against cancer. Front Oncol. 5:472015.PubMed/NCBI
34	Moriya S, Komatsu S, Yamasaki K, Kawai Y, Kokuba H, Hirota A, Che XF, Inazu M, Gotoh A, Hiramoto M, et al: Targeting the integrated networks of aggresome formation, proteasome, and autophagy potentiates ER stress-mediated cell death in multiple myeloma cells. Int J Oncol. 46:474–486. 2015.
35	Komatsu S, Moriya S, Che XF, Yokoyama T, Kohno N and Miyazawa K: Combined treatment with SAHA, bortezomib, and clarithromycin for concomitant targeting of aggresome formation and intracellular proteolytic pathways enhances ER stress-mediated cell death in breast cancer cells. Biochem Biophys Res Commun. 437:41–47. 2013. View Article : Google Scholar : PubMed/NCBI
36	Yu L, McPhee CK, Zheng L, Mardones GA, Rong Y, Peng J, Mi N, Zhao Y, Liu Z, Wan F, et al: Termination of autophagy and reformation of lysosomes regulated by mTOR. Nature. 465:942–946. 2010. View Article : Google Scholar : PubMed/NCBI
37	Zaarur N, Meriin AB, Bejarano E, Xu X, Gabai VL, Cuervo AM and Sherman MY: Proteasome failure promotes positioning of lysosomes around the aggresome via local block of microtubule-dependent transport. Mol Cell Biol. 34:1336–1348. 2014. View Article : Google Scholar : PubMed/NCBI
38	Mackeh R, Perdiz D, Lorin S, Codogno P and Poüs C: Autophagy and microtubules - new story, old players. J Cell Sci. 126:1071–1080. 2013. View Article : Google Scholar : PubMed/NCBI
39	Olzmann JA, Li L and Chin LS: Aggresome formation and neurodegenerative diseases: Therapeutic implications. Curr Med Chem. 15:47–60. 2008. View Article : Google Scholar : PubMed/NCBI
40	Nawrocki ST, Carew JS, Pino MS, Highshaw RA, Andtbacka RH, Dunner K Jr, Pal A, Bornmann WG, Chiao PJ, Huang P, et al: Aggresome disruption: A novel strategy to enhance bortezomib-induced apoptosis in pancreatic cancer cells. Cancer Res. 66:3773–3781. 2006. View Article : Google Scholar : PubMed/NCBI
41	Helfand BT, Chang L and Goldman RD: Intermediate filaments are dynamic and motile elements of cellular architecture. J Cell Sci. 117:133–141. 2004. View Article : Google Scholar
42	Perlson E, Michaelevski I, Kowalsman N, Ben-Yaakov K, Shaked M, Seger R, Eisenstein M and Fainzilber M: Vimentin binding to phosphorylated Erk sterically hinders enzymatic dephosphorylation of the kinase. J Mol Biol. 364:938–944. 2006. View Article : Google Scholar : PubMed/NCBI
43	Pérez-Sala D, Oeste CL, Martínez AE, Carrasco MJ, Garzón B and Cañada FJ: Vimentin filament organization and stress sensing depend on its single cysteine residue and zinc binding. Nat Commun. 6:72872015. View Article : Google Scholar : PubMed/NCBI