Inositol 1,4,5‑trisphosphate receptor type 3 is involved in resistance to apoptosis and maintenance of human hepatocellular carcinoma

Dos Santos,Marcone Loiola; França,Andressa; Lima Filho,Antônio Carlos Melo; Florentino,Rodrigo M.; Diniz,Paulo Henrique; Lemos,Fernanda Oliveira; Gonçalves,Carlos Alberto Xavier; Coelho,Vitor Lima; Lima,Cristiano Xavier; Foureaux,Giselle; Nathanson,Michael H.; Vidigal,Paula Vieira Teixeira; Leite,M. Fátima

doi:10.3892/ol.2021.13150

Inositol 1,4,5‑trisphosphate receptor type 3 is involved in resistance to apoptosis and maintenance of human hepatocellular carcinoma

Authors:
- Marcone Loiola Dos Santos
- Andressa França
- Antônio Carlos Melo Lima Filho
- Rodrigo M. Florentino
- Paulo Henrique Diniz
- Fernanda Oliveira Lemos
- Carlos Alberto Xavier Gonçalves
- Vitor Lima Coelho
- Cristiano Xavier Lima
- Giselle Foureaux
- Michael H. Nathanson
- Paula Vieira Teixeira Vidigal
- M. Fátima Leite
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Affiliations: Department of Morphology, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais 31270‑901, Brazil, Department of Physiology and Biophysics, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais 31270‑901, Brazil, Department of Internal Medicine, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais 31270‑901, Brazil, Coordination of Biotechnology, SENAI's Innovation Institute for Biosynthetics and Fibers, SENAI CETIQT, Rio de Janeiro 20961‑020, Brazil, Department of Surgery, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais 30 130‑100, Brazil, Section of Digestive Diseases, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06520‑8056, USA, Department of Pathological Anatomy and Forensic Medicine of Hospital das Clínicas, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais 30 130‑100, Brazil
Published online on: November 25, 2021 https://doi.org/10.3892/ol.2021.13150
Article Number: 32
Copyright: © Dos Santos et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The expression of the inositol 1,4,5‑trisphosphate receptor type 3 (ITRP3) in hepatocytes is a common event in the pathogenesis of hepatocellular carcinoma (HCC), regardless of the type of underlying liver disease. However, it is not known whether ITPR3 expression in hepatocytes is involved in tumor maintenance. The aim of the present study was to determine whether there is an association between ITPR3 expression and clinical and morphological parameters using HCC samples obtained from liver explants from patients (n=53) with different etiologies of underlying chronic liver disease (CLD). ITPR3 expression, mitosis and apoptosis were analyzed in human liver samples by immunohistochemistry. Clinical and event‑free survival data were combined to assess the relationship between ITPR3 and liver cancer growth in patients. RNA sequencing analysis was performed to identify apoptotic genes altered by ITPR3 expression in a liver tumor cell line. ITPR3 was highly expressed in HCC tumor cells relative to adjacent CLD tissue and healthy livers. There was an inverse correlation between ITPR3 expression and mitotic and apoptotic indices in HCC, suggesting that ITPR3 contributed to the maintenance of HCC by promoting resistance to apoptosis. This was confirmed by the upregulation of CTSB, CHOP and GADD45, genes involved in the apoptotic pathway in HCC. The expression of ITPR3 in the liver may be a promising prognostic marker of HCC.

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1	Agni RM: Diagnostic histopathology of hepatocellular carcinoma: A case-based review. Semin Diagn Pathol. 34:126–137. 2017. View Article : Google Scholar : PubMed/NCBI
2	Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A and Bray F: Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 71:209–249. 2021. View Article : Google Scholar : PubMed/NCBI
3	Forner A, Reig M and Bruix J: Hepatocellular carcinoma. Lancet. 391:1301–1314. 2018. View Article : Google Scholar : PubMed/NCBI
4	Bodzin AS, Lunsford KE, Markovic D, Harlander-Locke MP, Busuttil RW and Agopian VG: Predicting mortality in patients developing recurrent hepatocellular carcinoma after liver transplantation: Impact of treatment modality and recurrence characteristics. Ann Surg. 266:118–125. 2017. View Article : Google Scholar : PubMed/NCBI
5	Villanueva A: Hepatocellular carcinoma. N Engl J Med. 380:1450–1462. 2019. View Article : Google Scholar : PubMed/NCBI
6	Amaya MJ and Nathanson MH: Calcium signaling in the liver. Compr Physiol. 3:515–539. 2013. View Article : Google Scholar : PubMed/NCBI
7	Patel S, Joseph SK and Thomas AP: Molecular properties of inositol 1,4,5-trisphosphate receptors. Cell Calcium. 25:247–264. 1999. View Article : Google Scholar : PubMed/NCBI
8	Wojcikiewicz RJ: Type I, II, and III inositol 1,4,5-trisphosphate receptors are unequally susceptible to down-regulation and are expressed in markedly different proportions in different cell types. J Biol Chem. 270:11678–11683. 1995. View Article : Google Scholar : PubMed/NCBI
9	Yule DI, Ernst SA, Ohnishi H and Wojcikiewicz RJ: Evidence that zymogen granules are not a physiologically relevant calcium pool. Defining the distribution of inositol 1,4,5-trisphosphate receptors in pancreatic acinar cells. J Biol Chem. 272:9093–9098. 1997. View Article : Google Scholar : PubMed/NCBI
10	Cruz LN, Guerra MT, Kruglov E, Mennone A, Garcia CR, Chen J and Nathanson MH: Regulation of multidrug resistance-associated protein 2 by calcium signaling in mouse liver. Hepatology. 52:327–337. 2010. View Article : Google Scholar : PubMed/NCBI
11	Hirata K, Pusl T, O'Neill AF, Dranoff JA and Nathanson MH: The type II inositol 1,4,5-trisphosphate receptor can trigger Ca²⁺ waves in rat hepatocytes. Gastroenterology. 122:1088–1100. 2002. View Article : Google Scholar : PubMed/NCBI
12	Dufour JF, Lüthi M, Forestier M and Magnino F: Expression of inositol 1,4,5-trisphosphate receptor isoforms in rat cirrhosis. Hepatology. 30:1018–1026. 1999. View Article : Google Scholar : PubMed/NCBI
13	Lemos FO, França A, Lima Filho ACM, Florentino RM, Santos ML, Missiaggia DG, Rodrigues GOL, Dias FF, Souza Passos IB, Teixeira MM, et al: Molecular mechanism for protection against liver failure in human yellow fever infection. Hepatol Commun. 4:657–669. 2020. View Article : Google Scholar : PubMed/NCBI
14	Lima Filho ACM, França A, Florentino RM, Dos Santos ML, de Oliveira Lemos F, Missiaggia DG, Fonseca RC, Gustavo Oliveira A, Ananthanarayanan M, Guerra MT, et al: Inositol 1,4,5-trisphosphate receptor type 3 plays a protective role in hepatocytes during hepatic ischemia-reperfusion injury. Cell Calcium. 91:1022642020. View Article : Google Scholar : PubMed/NCBI
15	Leite MF, Thrower EC, Echevarria W, Koulen P, Hirata K, Bennett AM, Ehrlich BE and Nathanson MH: Nuclear and cytosolic calcium are regulated independently. Proc Natl Acad Sci USA. 100:2975–2980. 2003. View Article : Google Scholar : PubMed/NCBI
16	Guerra MT, Florentino RM, França A, Lima Filho AC, Dos Santos ML, Fonseca RC, Lemos FO, Fonseca MC, Kruglov E, Mennone A, et al: Expression of the type 3 InsP₃ receptor is a final common event in the development of hepatocellular carcinoma. Gut. 68:1676–1687. 2019. View Article : Google Scholar : PubMed/NCBI
17	Ueasilamongkol P, Khamphaya T, Guerra MT, Rodrigues MA, Gomes DA, Kong Y, Wei W, Jain D, Trampert DC, Ananthanarayanan M, et al: Type 3 inositol 1,4,5-trisphosphate receptor is increased and enhances malignant properties in cholangiocarcinoma. Hepatology. 71:583–599. 2020. View Article : Google Scholar : PubMed/NCBI
18	Shibao K, Fiedler MJ, Nagata J, Minagawa N, Hirata K, Nakayama Y, Iwakiri Y, Nathanson MH and Yamaguchi K: The type III inositol 1,4,5-trisphosphate receptor is associated with aggressiveness of colorectal carcinoma. Cell Calcium. 48:315–323. 2010. View Article : Google Scholar : PubMed/NCBI
19	Kuchay S, Giorgi C, Simoneschi D, Pagan J, Missiroli S, Saraf A, Florens L, Washburn MP, Collazo-Lorduy A, Castillo-Martin M, et al: PTEN counteracts FBXL2 to promote IP3R3- and Ca²⁺-mediated apoptosis limiting tumour growth. Nature. 546:554–558. 2017. View Article : Google Scholar : PubMed/NCBI
20	Bononi A, Giorgi C, Patergnani S, Larson D, Verbruggen K, Tanji M, Pellegrini L, Signorato V, Olivetto F, Pastorino S, et al: BAP1 regulates IP3R3-mediated Ca²⁺ flux to mitochondria suppressing cell transformation. Nature. 546:549–553. 2017. View Article : Google Scholar : PubMed/NCBI
21	Fonseca MC, França A, Florentino RM, Fonseca RC, Lima Filho AC, Vidigal PT, Oliveira AG, Dubuquoy L, Nathanson MH and Leite MF: Cholesterol-enriched membrane microdomains are needed for insulin signaling and proliferation in hepatic cells. Am J Physiol Gastrointest Liver Physiol. 315:G80–G94. 2018. View Article : Google Scholar : PubMed/NCBI
22	Baak JP: Mitosis counting in tumors. Hum Pathol. 21:683–685. 1990. View Article : Google Scholar : PubMed/NCBI
23	Ha SY, Choi M, Lee T and Park CK: The prognostic role of mitotic index in hepatocellular carcinoma patients after curative hepatectomy. Cancer Res Treat. 48:180–189. 2016. View Article : Google Scholar : PubMed/NCBI
24	Haratake J, Takeda S, Kasai T, Nakano S and Tokui N: Predictable factors for estimating prognosis of patients after resection of hepatocellular carcinoma. Cancer. 72:1178–1183. 1993. View Article : Google Scholar : PubMed/NCBI
25	Nanashima A, Tanaka K, Yamaguchi H, Shibasaki S, Morino S, Yoshinaga M, Sawai T, Nakagoe T and Ayabe H: Fibrosis and inflammatory activity in noncancerous tissue and mitotic index of cancer tissue in patients with hepatocellular carcinoma: Relationship to clinicopathological factors and prognosis after hepatic resection. Dig Dis Sci. 48:1517–1522. 2003. View Article : Google Scholar : PubMed/NCBI
26	Osório FM, Vidigal PV, Ferrari TC, Lima AS, Lauar GM and Couto CA: Histologic grade and mitotic index as predictors of microvascular invasion in hepatocellular carcinoma. Exp Clin Transplant. 13:421–425. 2015.PubMed/NCBI
27	Ouchi K, Sugawara T, Ono H, Fujiya T, Kamiyama Y, Kakugawa Y, Mikuni J, Yamanami H, Komatsu S and Horikoshi A: Mitotic index is the best predictive factor for survival of patients with resected hepatocellular carcinoma. Dig Surg. 17:42–48. 2000. View Article : Google Scholar : PubMed/NCBI
28	Bolger AM, Lohse M and Usadel B: Trimmomatic: A flexible trimmer for illumina sequence data. Bioinformatics. 30:2114–2120. 2014. View Article : Google Scholar : PubMed/NCBI
29	Patro R, Duggal G, Love MI, Irizarry RA and Kingsford C: Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods. 14:417–419. 2017. View Article : Google Scholar : PubMed/NCBI
30	Howe KL, Achuthan P, Allen J, Allen J, Alvarez-Jarreta J, Amode MR, Armean IM, Azov AG, Bennett R, Bhai J, et al: Ensembl 2021. Nucleic Acids Res. 49D:D884–D891. 2021. View Article : Google Scholar : PubMed/NCBI
31	Love MI, Huber W and Anders S: Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 15:5502014. View Article : Google Scholar : PubMed/NCBI
32	Kucukural A, Yukselen O, Ozata DM, Moore MJ and Garber M: DEBrowser: Interactive differential expression analysis and visualization tool for count data. BMC Genomics. 20:62019. View Article : Google Scholar : PubMed/NCBI
33	Kanehisa M and Goto S: KEGG: Kyoto encyclopedia of genes and genomes. Nucleic Acids Res. 28:27–30. 2000. View Article : Google Scholar : PubMed/NCBI
34	UniProt Consortium: UniProt: The universal protein knowledgebase in 2021. Nucleic Acids Resy. 49D:D480–D489. 2021.PubMed/NCBI
35	Kanehisa M and Sato Y: KEGG Mapper for inferring cellular functions from protein sequences. Protein Sci. 29:28–35. 2020. View Article : Google Scholar : PubMed/NCBI
36	Babicki S, Arndt D, Marcu A, Liang Y, Grant JR, Maciejewski A and Wishart DS: Heatmapper: Web-enabled heat mapping for all. Nucleic Acids Res. 44W:W147–W153. 2016. View Article : Google Scholar : PubMed/NCBI
37	Edmondson HA and Steiner PE: Primary carcinoma of the liver: A study of 100 cases among 48,900 necropsies. Cancer. 7:462–503. 1954. View Article : Google Scholar : PubMed/NCBI
38	Ruan J, Zheng H, Rong X, Rong X, Zhang J, Fang W, Zhao P and Luo R: Over-expression of cathepsin B in hepatocellular carcinomas predicts poor prognosis of HCC patients. Mol Cancer. 15:172016. View Article : Google Scholar : PubMed/NCBI
39	Scaiewicz V, Nahmias A, Chung RT, Mueller T, Tirosh B and Shibolet O: CCAAT/enhancer-binding protein homologous (CHOP) protein promotes carcinogenesis in the DEN-induced Hepatocellular carcinoma model. PLoS One. 8:e810652013. View Article : Google Scholar : PubMed/NCBI
40	Liebermann DA and Hoffman B: Gadd45 in stress signaling. J Mol Signal. 3:152008. View Article : Google Scholar : PubMed/NCBI
41	Qiu W, David D, Zhou B, Chu PG, Zhang B, Wu M, Xiao J, Han T, Zhu Z, Wang T, et al: Down-regulation of growth arrest DNA damage-inducible gene 45beta expression is associated with human hepatocellular carcinoma. Am J Pathol. 162:1961–1974. 2003. View Article : Google Scholar : PubMed/NCBI
42	Llovet JM, Peña CE, Lathia CD, Shan M, Meinhardt G and Bruix J; SHARP Investigators Study Group, : Plasma biomarkers as predictors of outcome in patients with advanced hepatocellular carcinoma. Clin Cancer Res. 18:2290–2300. 2012. View Article : Google Scholar : PubMed/NCBI
43	Alves RC, Alves D, Guz B, Matos C, Viana M, Harriz M, Terrabuio D, Kondo M, Gampel O and Polletti P: Advanced hepatocellular carcinoma. Review of targeted molecular drugs. Ann Hepatol. 10:21–27. 2011. View Article : Google Scholar : PubMed/NCBI
44	Ho DW, Lo RC, Chan LK and Ng IO: Molecular pathogenesis of hepatocellular carcinoma. Liver Cancer. 5:290–302. 2016. View Article : Google Scholar : PubMed/NCBI
45	Lu LC, Hsu CH, Hsu C and Cheng AL: Tumor heterogeneity in hepatocellular carcinoma: Facing the challenges. Liver Cancer. 5:128–138. 2016. View Article : Google Scholar : PubMed/NCBI
46	Hoshida Y, Nijman SM, Kobayashi M, Chan JA, Brunet JP, Chiang DY, Villanueva A, Newell P, Ikeda K, Hashimoto M, et al: Integrative transcriptome analysis reveals common molecular subclasses of human hepatocellular carcinoma. Cancer Res. 69:7385–7392. 2009. View Article : Google Scholar : PubMed/NCBI
47	Chiang DY, Villanueva A, Hoshida Y, Peix J, Newell P, Minguez B, LeBlanc AC, Donovan DJ, Thung SN, Solé M, et al: Focal gains of VEGFA and molecular classification of hepatocellular carcinoma. Cancer Res. 68:6779–6788. 2008. View Article : Google Scholar : PubMed/NCBI
48	Villanueva A, Hoshida Y, Battiston C, Tovar V, Sia D, Alsinet C, Cornella H, Liberzon A, Kobayashi M, Kumada H, et al: Combining clinical, pathology, and gene expression data to predict recurrence of hepatocellular carcinoma. Gastroenterology. 140:1501–1512.e2. 2011. View Article : Google Scholar : PubMed/NCBI
49	Lee JS, Chu IS, Heo J, Calvisi DF, Sun Z, Roskams T, Durnez A, Demetris AJ and Thorgeirsson SS: Classification and prediction of survival in hepatocellular carcinoma by gene expression profiling. Hepatology. 40:667–676. 2004. View Article : Google Scholar : PubMed/NCBI
50	Zucman-Rossi J, Villanueva A, Nault JC and Llovet JM: Genetic landscape and biomarkers of hepatocellular carcinoma. Gastroenterology. 149:1226–1239.e4. 2015. View Article : Google Scholar : PubMed/NCBI
51	Masuzaki R and Omata M: Screening program in high-risk populations. In: Hepatocellular Carcinoma. (2nd edition). Mcmasters KM and Vauthey JN: Springer. (New York, NY). 55–68. 2011.
52	Bloom HJ and Richardson WW: Histological grading and prognosis in breast cancer; a study of 1409 cases of which 359 have been followed for 15 years. Br J Cancer. 11:359–377. 1957. View Article : Google Scholar : PubMed/NCBI
53	Vang R, Shih IM and Kurman RJ: Ovarian low-grade and high-grade serous carcinoma: Pathogenesis, clinicopathologic and molecular biologic features, and diagnostic problems. Adv Anat Patht. 16:267–282. 2009. View Article : Google Scholar : PubMed/NCBI
54	Suehiro T, Matsumata T, Itasaka H, Yamamoto K, Kawahara N and Sugimachi K: Clinicopathologic features and prognosis of resected hepatocellular carcinomas of varied sizes with special reference to proliferating cell nuclear antigen. Cancer. 76:399–405. 1995. View Article : Google Scholar : PubMed/NCBI
55	Okada S, Shimada K, Yamamoto J, Takayama T, Kosuge T, Yamasaki S, Sakamoto M and Hirohashi S: Predictive factors for postoperative recurrence of hepatocellular carcinoma. Gastroenterology. 106:1618–1624. 1994. View Article : Google Scholar : PubMed/NCBI
56	Ruà S, Comino A, Fruttero A, Torchio P, Bouzari H, Taraglio S, Torchio B and Capussotti L: Flow cytometric DNA analysis of cirrhotic liver cells in patients with hepatocellular carcinoma can provide a new prognostic factor. Cancer. 78:1195–1202. 1996. View Article : Google Scholar : PubMed/NCBI
57	Soini Y, Virkajärvi N, Lehto VP and Pääkkö P: Hepatocellular carcinomas with a high proliferation index and a low degree of apoptosis and necrosis are associated with a shortened survival. Br J Cancer. 73:1025–1030. 1996. View Article : Google Scholar : PubMed/NCBI
58	Tannapfel A, Geissler F, Köckerling F, Katalinic A, Hauss J and Wittekind C: Apoptosis and proliferation in relation to histopathological variables and prognosis in hepatocellular carcinoma. J Pathol. 187:439–445. 1999. View Article : Google Scholar : PubMed/NCBI
59	Alisi A, Mele R, Spaziani A, Tavolaro S, Palescandolo E and Balsano C: Thr 446 phosphorylation of PKR by HCV core protein deregulates G2/M phase in HCC cells. J Cell Physiol. 205:25–31. 2005. View Article : Google Scholar : PubMed/NCBI
60	Clemens DL, Calisto LE, Sorrell MF and Tuma DJ: Ethanol metabolism results in a G2/M cell-cycle arrest in recombinant Hep G2 cells. Hepatology. 38:385–393. 2003. View Article : Google Scholar : PubMed/NCBI
61	Juríková M, Danihel L, Polák Š and Varga I: Ki67, PCNA, and MCM proteins: Markers of proliferation in the diagnosis of breast cancer. Acta Histochem. 118:544–552. 2016. View Article : Google Scholar : PubMed/NCBI
62	Michalopoulos GK: Liver regeneration after partial hepatectomy: Critical analysis of mechanistic dilemmas. Am J Pathol. 176:2–13. 2010. View Article : Google Scholar : PubMed/NCBI
63	Mangla A, Guerra MT and Nathanson MH: Type 3 inositol 1,4,5-trisphosphate receptor: A calcium channel for all seasons. Cell Calcium. 85:1021322020. View Article : Google Scholar : PubMed/NCBI
64	Chikka MR, McCabe DD, Tyra HM and Rutkowski DT: C/EBP homologous protein (CHOP) contributes to suppression of metabolic genes during endoplasmic reticulum stress in the liver. J Biol Chem. 288:4405–4415. 2013. View Article : Google Scholar : PubMed/NCBI
65	Tian J and Locker J: Gadd45 in the liver: Signal transduction and transcriptional mechanisms. In: Gadd45 Stress Sensor Genes. Liebermann DA and Hoffman B: Springer. (New York, NY). 69–80. 2013.
66	Xia H, Lee KW, Chen J, Kong SN, Sekar K, Deivasigamani A, Seshachalam VP, Goh BKP, Ooi LL and Hui KM: Simultaneous silencing of ACSL4 and induction of GADD45B in hepatocellular carcinoma cells amplifies the synergistic therapeutic effect of aspirin and sorafenib. Cell Death Discov. 3:170582017. View Article : Google Scholar : PubMed/NCBI