Inhibition of microtubule-associated protein 1 light chain 3B via small-interfering RNA or 3-methyladenine impairs hypoxia-induced HO8910PM and HO8910 epithelial ovarian cancer cell migration and invasion and is associated with RhoA and alterations of the actin cytoskeleton

Tang,Zhongyuan; Zhang,Ning; Di,Wen; Li,Weiping

doi:10.3892/or.2015.3742

Inhibition of microtubule-associated protein 1 light chain 3B via small-interfering RNA or 3-methyladenine impairs hypoxia-induced HO8910PM and HO8910 epithelial ovarian cancer cell migration and invasion and is associated with RhoA and alterations of the actin cytoskeleton

Authors:
- Zhongyuan Tang
- Ning Zhang
- Wen Di
- Weiping Li
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Affiliations: Department of Obstetrics and Gynecology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, P.R. China
Published online on: January 20, 2015 https://doi.org/10.3892/or.2015.3742
Pages: 1411-1417

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Abstract

Expression of microtubule‑associated protein 1 light chain 3 (LC3) is correlated with poor prognosis in many human cancers. Hypoxia induces LC3 expression and is an essential characteristic of epithelial ovarian cancer (EOC). The aim of the present study was to elucidate the mechanism by which LC3 facilitates EOC cell migration and invasion under conditions of hypoxia. The effects of LC3B inhibition under hypoxic conditions on migration, invasion, and adhesion in HO8910PM and HO8910 EOC cell lines were investigated. LC3B inhibition was achieved by small‑interfering RNA (siRNA) targeting LC3B or by treatment with 3‑methyladenine (3‑MA). Cell migration, invasion and adhesion and the arrangement of the cytoskeleton were determined by Transwell migration assays and rhodamine phalloidin staining. Western blot analysis was performed to evaluate the expression level of LC3B and the expression and activity of ras homolog gene family member A (RhoA). Increased LC3B expression was associated with HO8910PM and HO8910 cell migration and invasion promoted under hypoxic conditions. LC3B siRNA and 3‑MA treatment each attenuated hypoxia‑induced LC3B expression, along with migration and invasion, and this was associated with a decrease in RhoA expression and disorganization of the actin cytoskeleton. LC3B may promote the migration and invasion of EOC cells by affecting the cytoskeleton via the RhoA pathway. In addition, LC3B may be a marker of tumor hypoxia and/or metastasis in EOC cells.

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1	Davidson B, Reich R, Trope CG, Wang TL and Shih IeM: New determinates of disease progression and outcome in metastatic ovarian carcinoma. Histol Histopathol. 25:1591–1609. 2010.PubMed/NCBI
2	Bagnato A, Spinella F and Rosanò L: Emerging role of the endothelin axis in ovarian tumor progression. Endocr Relat Cancer. 12:761–772. 2005. View Article : Google Scholar : PubMed/NCBI
3	Itamochi H: Targeted therapies in epithelial ovarian cancer: molecular mechanisms of action. World J Biol Chem. 1:209–220. 2010. View Article : Google Scholar
4	Opipari AW Jr, Tan L, Boitano AE, Sorenson DR, Aurora A and Liu JR: Resveratrol-induced autophagocytosis in ovarian cancer cells. Cancer Res. 64:696–703. 2004. View Article : Google Scholar : PubMed/NCBI
5	Simonin K, Brotin E, Dufort S, Dutoit S, Goux D, N’diaye M, Denoyelle C, Gauduchon P and Poulain L: Mcl-1 is an important determinant of the apoptotic response to the BH3-mimetic molecule HA14-1 in cisplatin-resistant ovarian carcinoma cells. Mol Cancer Ther. 8:3162–3170. 2009. View Article : Google Scholar : PubMed/NCBI
6	Saad AF, Hu W and Sood AK: Microenvironment and pathogenesis of epithelial ovarian cancer. Horm Cancer. 1:277–290. 2010. View Article : Google Scholar
7	Zhang J, Yang Z, Xie L, Xu L, Xu D and Liu X: Statins, autophagy and cancer metastasis. Int J Biochem Cell Biol. 45:745–752. 2013. View Article : Google Scholar
8	Bhoopathi P, Gondi CS, Gujrati M, Dinh DH and Lakka SS: SPARC mediates Src-induced disruption of actin cytoskeleton via inactivation of small GTPases Rho-Rac-Cdc42. Cell Signal. 23:978–1987. 2011. View Article : Google Scholar
9	Du H, Yang W, Chen L, Shen B, Peng C, Li H, Ann DK, Yen Y and Qiu W: Emerging role of autophagy during ischemia-hypoxia and reperfusion in hepatocellular carcinoma. Int J Oncol. 40:2049–2057. 2012.PubMed/NCBI
10	Sun Y, Liu JH, Sui YX, Jin L, Yang Y, Lin SM and Shi H: Beclin1 overexpression inhibits proliferation, invasion and migration of CaSki cervical cancer cells. Asian Pac J Cancer Prev. 12:1269–1273. 2011.
11	Rouschop KM and Wouters BG: Regulation of autophagy through multiple independent hypoxic signaling pathways. Curr Mol Med. 9:417–424. 2009. View Article : Google Scholar : PubMed/NCBI
12	Schlie K, Spowart JE, Hughson LR, Townsend KN and Lum JJ: When cells suffocate: autophagy in cancer and immune cells under low oxygen. Int J Cell Biol. 2011:470–597. 2011. View Article : Google Scholar
13	Indelicato M, Pucci B, Schito L, Reali V, Aventaggiato M, Mazzarino MC, Stivala F, Fini M, Russo MA and Tafani M: Role of hypoxia and autophagy in MDA-MB-231 invasiveness. J Cell Physiol. 223:359–368. 2010.PubMed/NCBI
14	Tanida I, Ueno T and Kominami E: LC3 conjugation system in mammalian autophagy. Int J Biochem Cell Biol. 36:2503–2518. 2004. View Article : Google Scholar : PubMed/NCBI
15	Martinet W, De Meyer GR, Andries L, Herman AG and Kockx MM: In situ detection of starvation-induced autophagy. J Histochem Cytochem. 54:85–96. 2006. View Article : Google Scholar
16	Peracchio C, Alabiso O, Valente G and Isidoro C: Involvement of autophagy in ovarian cancer: a working hypothesis. J Ovarian Res. 5:222012. View Article : Google Scholar : PubMed/NCBI
17	Hu YL, DeLay M, Jahangiri A, Molinaro AM, Rose SD, Carbonell WS and Aghi MK: Hypoxia-induced autophagy promotes tumor cell survival and adaptation to antiangiogenic treatment in glioblastoma. Cancer Res. 72:1773–1783. 2012. View Article : Google Scholar : PubMed/NCBI
18	Aguilera MO, Berón W and Colombo MI: The actin cytoskeleton participates in the early events of autophagosome formation upon starvation induced autophagy. Autophagy. 8:1590–1603. 2012. View Article : Google Scholar : PubMed/NCBI
19	Pankiv S, Alemu EA, Brech A, Bruun JA, Lamark T, Overvatn A, Bjørkøy G and Johansen T: FYCO1 is a Rab7 effector that binds to LC3 and PI3P to mediate microtubule plus end-directed vesicle transport. J Cell Biol. 188:253–269. 2010. View Article : Google Scholar : PubMed/NCBI
20	Rapisarda A, Uranchimeg B, Scudiero DA, Selby M, Sausville EA, Shoemaker RH and Melillo G: Identification of small molecule inhibitors of hypoxia-inducible factor 1 transcriptional activation pathway. Cancer Res. 62:4316–324. 2002.PubMed/NCBI
21	Lazova R, Camp RL, Klump V, Siddiqui SF, Amaravadi RK and Pawelek JM: Punctate LC3B expression is a common feature of solid tumors and associated with proliferation, metastasis, and poor outcome. Clin Cancer Res. 18:370–379. 2012. View Article : Google Scholar
22	Sato K, Tsuchihara K, Fujii S, Sugiyama M, Goya T, Atomi Y, Ueno T, Ochiai A and Esumi H: Autophagy is activated in colorectal cancer cells and contributes to the tolerance to nutrient deprivation. Cancer Res. 67:9677–9684. 2007. View Article : Google Scholar : PubMed/NCBI
23	Yoshioka A, Miyata H, Doki Y, Yamasaki M, Sohma I, Gotoh K, Takiguchi S, Fujiwara Y, Uchiyama Y and Monden M: LC3, an autophagosome marker, is highly expressed in gastrointestinal cancers. Int J Oncol. 33:461–468. 2008.PubMed/NCBI
24	Han C, Sun B, Wang W, Cai W, Lou D, Sun Y and Zhao X: Overexpression of microtubule-associated protein-1 light chain 3 is associated with melanoma metastasis and vasculogenic mimicry. Tohoku J Exp Med. 223:243–251. 2011. View Article : Google Scholar : PubMed/NCBI
25	Fujii S, Mitsunaga S, Yamazaki M, Hasebe T, Ishii G, Kojima M, Kinoshita T, Ueno T, Esumi H and Ochiai A: Autophagy is activated in pancreatic cancer cells and correlates with poor patient outcome. Cancer Sci. 99:1813–1819. 2008.PubMed/NCBI
26	Qin AP, Liu CF, Qin YY, Hong LZ, Xu M, Yang L, Liu J, Qin ZH and Zhang HL: Autophagy was activated in injured astrocytes and mildly decreased cell survival following glucose and oxygen deprivation and focal cerebral ischemia. Autophagy. 6:738–753. 2010. View Article : Google Scholar : PubMed/NCBI
27	Codogno P and Meijer AJ: Autophagy and signaling: their role in cell survival and cell death. Cell Death Differ. 12(Suppl 2): 1509–1518. 2005. View Article : Google Scholar : PubMed/NCBI
28	Myeku N and Figueiredo-Pereira ME: Dynamics of the degradation of ubiquitinated proteins by proteasomes and autophagy: association with sequestosome 1/p62. J Biol Chem. 286:22426–22440. 2011. View Article : Google Scholar : PubMed/NCBI
29	Song J, Qu Z, Guo X, Zhao Q, Zhao X, Gao L, Sun K, Shen F, Wu M and Wei L: Hypoxia-induced autophagy contributes to the chemoresistance of hepatocellular carcinoma cells. Autophagy. 5:1131–1144. 2009. View Article : Google Scholar : PubMed/NCBI
30	Kadandale P, Stender JD, Glass CK and Kiger AA: Conserved role for autophagy in Rho1-mediated cortical remodeling and blood cell recruitment. Proc Natl Acad Sci USA. 107:10502–10507. 2010. View Article : Google Scholar : PubMed/NCBI