Ultrastructural and immunofluorescent analysis of lithium effects on autophagy in hepatocellular carcinoma cells

Iuliia Taskaeva, Nataliya Bgatova


Background: Hepatocellular carcinoma (HCC) is one of the most malignant cancers worldwide. The role of autophagy in the НСС development and progression is controversial and are still not well known. It has been shown that lithium induces autophagy and apoptosis, affects on proliferation and survival of various human malignancies. In this study we estimated lithium`s effects on autophagy in HCC in vivo.

Materials and methods: Subcellular components and autophagic vacuoles were analyzed by transmission electron microscopy. The number of cells with LC3 and LAMP1 punctate was analyzed by double-immunofluorescence staining. Data are presented as mean ± standard deviation (SD). Mann–Whitney nonparametric tests were used to assess statistically significant differences at P < 0.05.  

Results: Lithium carbonate increased the numerical and volume density of autophagic vacuoles and autophagy marker LC3 beta. Therefore, lithium carbonate promotes enlarged autophagic vacuoles formation in HCC cells in vivo.

Conclusion: Extended and unresolved autophagy induces cancer cell death by stimulating autophagic cell death. Thus, lithium-mediated autophagy can be an attractive approach in HCC chemotherapy.

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Kroemer, G. and Levine, B. Autophagic cell death: the story of a misnomer. Nature reviews. Molecular cell biology. 2008;9(12):1004-10.

Kondo, Y., Kanzawa, T., Sawaya, R. and Kondo, S. The role of autophagy in cancer development and response to therapy. Nature reviews. Cancer, 2005;5(9):726-34.

Janku, F., McConkey, D.J., Hong, D.S. and Kurzrock, R. Autophagy as a target for anticancer therapy. Nature reviews. Clinical oncology. 2011;8(9):528-39.

Best, J., Schotten, C., Theysohn, J.M., Wetter, A., Müller, S., Radünz, S., et al. Novel implications in the treatment of hepatocellular carcinoma. Annals of gastroenterology. 2017;30:23-32.

Hartke, J., Johnson, M. and Ghabril, M. The diagnosis and treatment of hepatocellular carcinoma. Seminars in diagnostic pathology. 2015;34:153-159.

Lee, Y.J. and Jang, B.K. The role of autophagy in hepatocellular carcinoma. International journal of molecular sciences. 2015;16:26629-43.

Dash, S., Chava, S., Chandra, P.K., Aydin, Y., Balart, L.A. and Wu, T. Autophagy in hepatocellular carcinomas: from pathophysiology to therapeutic response. Hepatic medicine : evidence and research. 2016;8:9-20.

Liu, L., Liao, J.Z., He, X.X. and Li, P.Y. The role of autophagy in hepatocellular carcinoma: friend or foe. Oncotarget. 2017;8:57707-57722.

Washington, M.N., Suh, G., Orozco, A.F., Sutton, M.N., Yang, H., Wang, Y., et al. ARHI (DIRAS3)-mediated autophagy-associated cell death enhances chemosensitivity to cisplatin in ovarian cancer cell lines and xenografts. Cell death and disease. 2015. Available at: doi: 10.1038/cddis.2015.208.

Wang, Z., Liu, S., Ding, K., Ding, S., Li, C., Lu, J. et al. Silencing Livin induces apoptotic and autophagic cell death, increasing chemotherapeutic sensitivity to cisplatin of renal carcinoma cells. Tumor biology. 2016;37(11):15133-15143.

Zhang, C., Lei, J.L., Zhang, H., Xia, Y.Z., Yu, P., Yang, L. and Kong, L.Y. Calyxin Y sensitizes cisplatin-sensitive and resistant hepatocellular carcinoma cells to cisplatin through apoptotic and autophagic cell death via SCF βTrCP-mediated eEF2K degradation. Oncotarget. 2017;8(41):70595-70616.

Sarkar, S., Ravikumar, B., Floto, R.A. and Rubinsztein, D.C. Rapamycin and mTOR-independent autophagy inducers ameliorate toxicity of polyglutamine-expanded huntingtin and related proteinopathies. Cell death and differentiation. 2009;16:46-56.

Li, L., Song, H., Zhong, L., Yang, R., Yang, X.Q., Jiang, K.L. and Liu, B.Z. Lithium chloride promotes apoptosis in human leukemia NB4 cells by inhibiting glycogen synthase kinase-3 beta. International journal of medical sciences. 2015;12:805-10.

Zassadowski, F., Pokorna, K., Ferre, N., Guidez, F., Llopis, L., Chourbagi, O., et al. Lithium chloride antileukemic activity in acute promyelocytic leukemia is GSK-3 and MEK/ERK dependent. Leukemia. 2015;29:2277-84.

Kaledin, V.I., Zhukova, N.A., Nikolin, V.P., Popova, N.A., Beliaev, M.D., Baginskaya, N.V., et al. Hepatocarcinoma-29, a metastasizing transplantable mouse tumor inducing cachexia. Bulletin of experimental biology and medicine. 2009;148:903-8.

Klionsky, D.J., Abdelmohsen, K., Abe, A., Abedin, M.J., Abeliovich, H., Acevedo Arozena, A., et al. Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 2016;12:1-222.

Ávalos, Y., Canales, J., Bravo-Sagua, R., Criollo, A., Lavandero, S. and Quest, A.F. Tumor suppression and promotion by autophagy. BioMed research international. 2014. Available at: doi: 10.1155/2014/603980.

Janji, B., Viry, E., Moussay, E., Paggetti, J., Arakelian, T., Mgrditchian, T., et al. The multifaceted role of autophagy in tumor evasion from immune surveillance. Oncotarget. 2016;7:17591-607.

Kimmelman, A.C. and White, E. Autophagy and tumor metabolism. Cell metabolism. 2017;25:1037-1043.

Lorente, J., Velandia, C., Leal, J.A., Garcia-Mayea, Y., Lyakhovich, A., Kondoh, H. and LLeonart, M.E. The interplay between autophagy and tumorigenesis: exploiting autophagy as a means of anticancer therapy. Biological reviews of the Cambridge Philosophical Society. 2018;93:152-165.

White, E., Mehnert, J.M. and Chan, C.S. Autophagy, metabolism, and cancer. Clinical cancer research: an official journal of the American Association for Cancer Research. 2015;21:5037-46.

Pietrocola, F., Bravo-San Pedro, J.M., Galluzzi, L. and Kroemer, G. Autophagy in natural and therapy-driven anticancer immunosurveillance. Autophagy. 2017;13:2163-2170.

Pan, H., Chen, L., Xu, Y., Han, W., Lou, F., Fei, W., et al. Autophagy-associated immune responses and cancer immunotherapy. Oncotarget. 2016;7:21235-46.

Hu, L., Jiang, K., Ding, C. and Meng, S. Targeting autophagy for oncolytic immunotherapy. Biomedicines. 2017. Available at: doi: 10.3390/biomedicines5010005.

Zhong, Z., Sanchez-Lopez, E. and Karin, M. Autophagy, inflammation, and immunity: a troika governing cancer and its treatment. Cell. 2016;166:288-298.

Li, C.J., Liao, W.T., Wu, M.Y. and Chu, P.Y. New insights into the role of autophagy in tumor immune microenvironment. International journal of molecular sciences. 2017. Available at: doi: 10.3390/ijms18071566.

Motoi, Y., Shimada, K., Ishiguro, K. and Hattori, N. Lithium and autophagy. American Chemical Society chemical neuroscience. 2014;5:434-42.

Sarkar, S., Floto, R.A., Berger, Z., Imarisio, S., Cordenier, A., Pasco, M., et al. Lithium induces autophagy by inhibiting inositol monophosphatase. The Journal of cell biology. 2005;170:1101-11.

Sade, Y., Toker, L., Kara, N.Z., Einat, H., Rapoport, S., Moechars, D., et al. IP3 accumulation and/or inositol depletion: two downstream lithium's effects that may mediate its behavioral and cellular changes. Translational psychiatry. 2016. Available at: doi: 10.1038/tp.2016.217.

Vicencio, J.M., Ortiz, C., Criollo, A., Jones, A.W., Kepp, O., Galluzzi, L., et al. The inositol 1,4,5-trisphosphate receptor regulates autophagy through its interaction with Beclin 1. Cell death and differentiation. 2009;16:1006-17.

Fu, Y., Jiao, Y., Zheng, S., Liang, A. and Hu, F. Combination of lithium chloride and pEGFP-N1-BmK CT effectively decreases proliferation and migration of C6 glioma cells. Cytotechnology. 2016;68:197-202.

Gao, S., Li, S., Duan, X., Gu, Z., Ma, Z., Yuan, X., et al. Inhibition of glycogen synthase kinase 3 beta (GSK3β) suppresses the progression of esophageal squamous cell carcinoma by modifying STAT3 activity. Molecular carcinogenesis. 2017;56:2301-2316.

Wang, X., Luo, C., Cheng, X. and Lu, M. Lithium and an EPAC-specific inhibitor ESI-09 synergistically suppress pancreatic cancer cell proliferation and survival. Acta biochimica et biophysica Sinica. 2017;49:573-580.

Costabile, V., Duraturo, F., Delrio, P., Rega, D., Pace, U., Liccardo, R., et al. Lithium chloride induces mesenchymal to epithelial reverting transition in primary colon cancer cell cultures. International journal of oncology. 2015;46:1913-23.

Peixoto-da-Silva, J., Calgarotto, A.K, Rocha, K.R., Palmeira-Dos-Santos, C., Smaili, S.S., Pereira, G.J.S., et al. Lithium, a classic drug in psychiatry, improves nilotinib-mediated antileukemic effects. Biomedicine & pharmacotherapy. 2018;99:237-244.

O'Donovan, T.R., Rajendran, S., O'Reilly, S., O'Sullivan, G.C. and McKenna, S.L. Lithium modulates autophagy in esophageal and colorectal cancer cells and enhances the efficacy of therapeutic agents in vitro and in vivo. PLoS One. 2015. Available at: doi: 10.1371/journal.pone.0134676.

Trnski, D., Sabol, M., Gojević, A., Martinić, M., Ozretić, P., Musani, V., et al. GSK3β and Gli3 play a role in activation of Hedgehog-Gli pathway in human colon cancer - Targeting GSK3β downregulates the signaling pathway and reduces cell proliferation. Biochimica et biophysica acta. 2015;1852(12):2574-84.

Erdal, E., Ozturk, N., Cagatay, T., Eksioglu-Demiralp, E. and Ozturk, M. Lithium-mediated downregulation of PKB/Akt and cyclin E with growth inhibition in hepatocellular carcinoma cells. International journal of cancer. 2005;115:903-10.

Beurel, E., Blivet-Van Eggelpoël, M.J., Kornprobst, M., Moritz, S., Delelo, R., Paye, F., et al. Glycogen synthase kinase-3 inhibitors augment TRAIL-induced apoptotic death in human hepatoma cells. Biochemical pharmacology. 2009;77:54-65.

DOI (PDF (FULL TEXT)): http://dx.doi.org/10.22034/APJCB.2018.3.3.83


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