Overexpression of ribonuclease inhibitor induces autophagy in human colorectal cancer cells via the Akt/mTOR/ULK1 pathway

Tang,Ying; Ren,Feng; Cong,Xi; Kong,Ying; Tian,Yuxiang; Xu,Yuefei; Fan,Jianhui

doi:10.3892/mmr.2019.10030

Overexpression of ribonuclease inhibitor induces autophagy in human colorectal cancer cells via the Akt/mTOR/ULK1 pathway

Authors:
- Ying Tang
- Feng Ren
- Xi Cong
- Ying Kong
- Yuxiang Tian
- Yuefei Xu
- Jianhui Fan
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Affiliations: Department of Pathology, The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116023, P.R. China, Department of Biochemistry and Molecular Biology, Dalian Medical University, Dalian, Liaoning 116044, P.R. China
Published online on: March 14, 2019 https://doi.org/10.3892/mmr.2019.10030
Pages: 3519-3526
Copyright: © Tang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Ribonuclease inhibitor (RI), also termed angiogenin inhibitor, acts as the inhibitor of ribonucleolytic activity of RNase A and angiogenin. The expression of RI has been investigated in melanoma and bladder cancer cells. However, the precise role of RI in tumorigenesis, in addition to RI‑induced autophagy, remains poorly understood. In the present study, it was demonstrated that RI positively regulated autophagy in human colorectal cancer (CRC) cells as indicated by an increase in light chain 3 (LC3)‑II levels. Furthermore, RI regulated cell survival in HT29 cells. In addition, autophagy‑associated proteins, including beclin‑1 and autophagy‑related protein 13, were increased in response to RI‑induced autophagy, and the protein kinase B (Akt)/mechanistic target of rapamycin (mTOR)/Unc‑51 like autophagy activating kinase (ULK1) pathway may be involved in the activation of autophagy induced by RI overexpression. Taken together, the evidence of the present study indicated that the overexpression of RI induced ATG‑dependent autophagy in CRC cells via the Akt/mTOR/ULK1 pathway, suggesting that the upregulation of RI activity may constitute a novel approach for the treatment of human colorectal carcinoma.

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1	Siegel RL, Miller KD and Jemal A: Cancer statistics, 2016. CA Cancer J Clin. 66:7–30. 2016. View Article : Google Scholar : PubMed/NCBI
2	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
3	Ciombor KK, Wu C and Goldberg RM: Recent therapeutic advances in the treatment of colorectal cancer. Annu Rev Med. 66:83–95. 2015. View Article : Google Scholar : PubMed/NCBI
4	Kobe B and Deisenhofer J: Crystal structure of porcine ribonuclease inhibitor, a protein with leucine-rich repeats. Nature. 366:751–756. 1993. View Article : Google Scholar : PubMed/NCBI
5	McEwan PA, Scott PG, Bishop PN and Bella J: Structural correlations in the family of small leucine-rich repeat proteins and proteoglycans. J Struct Biol. 155:294–305. 2006. View Article : Google Scholar : PubMed/NCBI
6	Datta D, Samanta A, Dasgupta S and Pathak T: 3′-Oxo-, amino-, thio- and sulfone-acetic acid modified thymidines: Effect of increased acidity on ribonuclease A inhibition. Bioorg Med Chem. 21:4634–4645. 2013. View Article : Google Scholar : PubMed/NCBI
7	Dickson KA, Haigis MC and Raines RT: Ribonuclease inhibitor: Structure and function. Prog Nucleic Acid Res Mol Biol. 80:349–374. 2005. View Article : Google Scholar : PubMed/NCBI
8	Pan X, Xiong D, Yao X, Xin Y, Zhang L and Chen J: Up-regulating ribonuclease inhibitor inhibited epithelial-to-mesenchymal transition and metastasis in murine melanoma cells. Int J Biochem Cell Biol. 44:998–1008. 2012. View Article : Google Scholar : PubMed/NCBI
9	Yao X, Li D, Xiong DM, Li L, Jiang R and Chen JX: A novel role of ribonuclease inhibitor in regulation of epithelial-to-mesenchymal transition and ILK signaling pathway in bladder cancer cells. Cell Tissue Res. 353:409–423. 2013. View Article : Google Scholar : PubMed/NCBI
10	Chen JX, Gao Y, Liu JW, Tian YX, Zhao J and Cui XY: Antitumor effects of human ribonuclease inhibitor gene transfected on B16 melanoma cells. Int J Biochem Cell Biol. 37:1219–1231. 2005. View Article : Google Scholar : PubMed/NCBI
11	Tian YX, Wang DM and Cui XY: Analysis of gene expression of ribonuclease inhibitor in human breast cancer tissue. Ai Zheng. 23:269–272. 2004.(In Chinese). PubMed/NCBI
12	Tang Y, Liu P, Tian Y, Xu Y, Ren F, Cui X and Fan J: Overexpression of ribonuclease inhibitor defines good prognosis and suppresses proliferation and metastasis in human colorectal cancer cells via PI3K/AKT pathway. Clin Transl Oncol. 17:306–313. 2015. View Article : Google Scholar : PubMed/NCBI
13	Degtyarev M, De Mazière A, Orr C, Lin J, Lee BB, Tien JY, Prior WW, van Dijk S, Wu H, Gray DC, et al: Akt inhibition promotes autophagy and sensitizes PTEN-null tumors to lysosomotropic agents. J Cell Biol. 183:101–116. 2008. View Article : Google Scholar : PubMed/NCBI
14	Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y and Yoshimori T: LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 19:5720–5728. 2000. View Article : Google Scholar : PubMed/NCBI
15	Mizushima N, Levine B, Cuervo AM and Klionsky DJ: Autophagy fights disease through cellular self-digestion. Nature. 451:1069–1075. 2008. View Article : Google Scholar : PubMed/NCBI
16	White E and DiPaola RS: The double-edged sword of autophagy modulation in cancer. Clin Cancer Res. 15:5308–5316. 2009. View Article : Google Scholar : PubMed/NCBI
17	Ellington AA, Berhow M and Singletary KW: Induction of macroautophagy in human colon cancer cells by soybean B-group triterpenoid saponins. Carcinogenesis. 26:159–167. 2005. View Article : Google Scholar : PubMed/NCBI
18	Liang C, Feng P, Ku B, Dotan I, Canaani D, Oh BH and Jung JU: Autophagic and tumour suppressor activity of a novel Beclin1-binding protein UVRAG. Nat Cell Biol. 8:688–699. 2006. View Article : Google Scholar : PubMed/NCBI
19	Cheng Y, Ren X, Zhang Y, Patel R, Sharma A, Wu H, Robertson GP, Yan L, Rubin E and Yang JM: eEF-2 kinase dictates cross-talk between autophagy and apoptosis induced by Akt Inhibition, thereby modulating cytotoxicity of novel Akt inhibitor MK-2206. Cancer Res. 71:2654–2663. 2011. View Article : Google Scholar : PubMed/NCBI
20	Zhuang X, Lv M, Zhong Z, Zhang L, Jiang R and Chen J: Interplay between intergrin-linked kinase and ribonuclease inhibitor affects growth and metastasis of bladder cancer through signaling ILK pathways. J Exp Clin Cancer Res. 35:1302016. View Article : Google Scholar : PubMed/NCBI
21	Ji S, Lin W, Wang L, Ni Z, Jin F, Zha X and Fei G: Combined targeting of mTOR and Akt using rapamycin and MK-2206 in the treatment of tuberous sclerosis complex. J Cancer. 8:555–562. 2017. View Article : Google Scholar : PubMed/NCBI
22	Pizzo E, Sarcinelli C, Sheng J, Fusco S, Formiggini F, Netti P, Yu W, D'Alessio G and Hu GF: Ribonuclease/angiogenin inhibitor 1 regulates stress-induced subcellular localization of angiogenin to control growth and survival. J Cell Sci. 126:4308–4319. 2013. View Article : Google Scholar : PubMed/NCBI
23	Zhou JH, Tang XY, Zhao R, Wang H and Xia J: Effects of ribonuclease inhibitor on apoptosis and invasion of human breast cancer MDA-MB-231 cells. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 28:260–264. 2012.(In Chinese). PubMed/NCBI
24	Han K, Kim J and Choi M: Autophagy mediates phase transitions from cell death to life. Heliyon. 1:e000272015. View Article : Google Scholar : PubMed/NCBI
25	Carroll RG and Martin SJ: Autophagy in multiple myeloma: What makes you stronger can also kill you. Cancer Cell. 23:425–426. 2013. View Article : Google Scholar : PubMed/NCBI
26	Dickson KA, Kang DK, Kwon YS, Kim JC, Leland PA, Kim BM, Chang SI and Raines RT: Ribonuclease inhibitor regulates neovascularization by human angiogenin. Biochemistry. 48:3804–3806. 2009. View Article : Google Scholar : PubMed/NCBI
27	Peng Y, Li L, Huang M, Duan C, Zhang L and Chen J: Angiogenin interacts with ribonuclease inhibitor regulating PI3K/AKT/mTOR signaling pathway in bladder cancer cells. Cell Signal. 26:2782–2792. 2014. View Article : Google Scholar : PubMed/NCBI
28	Liu JL, Chang KC, Lo CC, Chu PY and Liu CH: Expression of autophagy-related protein Beclin-1 in malignant canine mammary tumors. BMC Vet Res. 9:752013. View Article : Google Scholar : PubMed/NCBI
29	Young PG, Passalacqua MJ, Chappell K, Llinas RJ and Bartel B: A facile forward-genetic screen for Arabidopsis autophagy mutants reveals twenty-one loss-of-function mutations disrupting six ATG genes. Autophagy. 1–19. 2019. View Article : Google Scholar
30	Nishida Y, Arakawa S, Fujitani K, Yamaguchi H, Mizuta T, Kanaseki T, Komatsu M, Otsu K, Tsujimoto Y and Shimizu S: Discovery of Atg5/Atg7-independent alternative macroautophagy. Nature. 461:654–658. 2009. View Article : Google Scholar : PubMed/NCBI
31	Qi S, Kim DJ, Stjepanovic G and Hurley JH: Structure of the human Atg13-Atg101 HORMA heterodimer: An interaction hub within the ULK1 complex. Structure. 23:1848–1857. 2015. View Article : Google Scholar : PubMed/NCBI
32	Levine B: Cell biology: Autophagy and cancer. Nature. 446:745–747. 2007. View Article : Google Scholar : PubMed/NCBI
33	Liu M, Zhao G, Zhang D, An W, Lai H, Li X, Cao S and Lin X: Active fraction of clove induces apoptosis via PI3K/Akt/mTOR-mediated autophagy in human colorectal cancer HCT-116 cells. Int J Oncol. 53:1363–1373. 2018.PubMed/NCBI
34	Yang L, Liu Y, Wang M, Qian Y, Dai X, Zhu Y, Chen J, Guo S and Hisamitsu T: Celastrus orbiculatus extract triggers apoptosis and autophagy via PI3K/Akt/mTOR inhibition in human colorectal cancer cells. Oncol Lett. 12:3771–3778. 2016. View Article : Google Scholar : PubMed/NCBI
35	Fan XJ, Wang Y, Wang L and Zhu M: Salidroside induces apoptosis and autophagy in human colorectal cancer cells through inhibition of PI3K/Akt/mTOR pathway. Oncol Rep. 36:3559–3567. 2016. View Article : Google Scholar : PubMed/NCBI
36	Kim J, Kundu M, Viollet B and Guan KL: AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol. 13:132–141. 2011. View Article : Google Scholar : PubMed/NCBI
37	Egan DF, Shackelford DB, Mihaylova MM, Gelino S, Kohnz RA, Mair W, Vasquez DS, Joshi A, Gwinn DM, Taylor R, et al: Phosphorylation of ULK1 (hATG1) by AMP-activated protein kinase connects energy sensing to mitophagy. Science. 331:456–461. 2011. View Article : Google Scholar : PubMed/NCBI
38	Lin Y, Kuang W, Wu B, Xie C, Liu C and Tu Z: IL-12 induces autophagy in human breast cancer cells through AMPK and the PI3K/Akt pathway. Mol Med Rep. 16:4113–4118. 2017. View Article : Google Scholar : PubMed/NCBI
39	Zhou X, Yue GG, Chan AM, Tsui SK, Fung KP, Sun H, Pu J and Lau CB: Eriocalyxin B, a novel autophagy inducer, exerts anti-tumor activity through the suppression of Akt/mTOR/p70S6K signaling pathway in breast cancer. Biochem Pharmacol. 142:58–70. 2017. View Article : Google Scholar : PubMed/NCBI
40	Mizushima N and Komatsu M: Autophagy: Renovation of cells and tissues. Cell. 147:728–741. 2011. View Article : Google Scholar : PubMed/NCBI
41	Green DR and Levine B: To be or not to be? How selective autophagy and cell death govern cell fate. Cell. 157:65–75. 2014. View Article : Google Scholar : PubMed/NCBI
42	Fujiwara K, Iwado E, Mills GB, Sawaya R, Kondo S and Kondo Y: Akt inhibitor shows anticancer and radiosensitizing effects in malignant glioma cells by inducing autophagy. Int J Oncol. 31:753–760. 2007.PubMed/NCBI