Increased angiopoietin-like protein 8 expression in colorectal carcinoma patients with liver metastasis

Abstract

Colorectal cancer (CRC) is the third most common cancer in men and the second most common in women, with a 5-year survival rate. The disease burden of CRC can be exacerbated by a sedentary lifestyle, lack of physical exercise, smoking, and obesity. Angiopoietin-like protein 8 (ANGPTL8), a liver hormone, plays a crucial role in lipid and glucose metabolism and is also implicated in the development of certain cancers. However, the basic profile and activity of this hormone in CRC with liver metastasis remains unknown. This study aims to establish the baseline expression of ANGPTL8 in tumors of CRC patients with and without liver metastasis. After histopathological screening and computed tomography scans, 41 patients were identified and included in this study, consisting of 20 CRC patients without liver metastasis and 21 with liver metastasis. Tumor samples from each group were compared to their respective adjacent normal tissues. Quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay were employed to assess the ANGPTL8 expression at both mRNA and protein levels. The results show that ANGPTL8 expression is higher in tumors of CRC patients with liver metastasis compared to those without liver metastasis. ANGPTL8 expression is significantly correlated with body mass index (BMI) (Pearson’s correlation coefficient = −0.367, p≤0.05) and could serve as a significant predictor of BMI (p≤0.05). These findings suggest that ANGPTL8 may enhance tumor development in CRC patients and is associated with body weight changes, contributing to weight loss symptoms during metastasis.

Keywords

ANGPTL8 / Body weight loss / Colorectal cancer / Liver metastasis

Funding

This study was supported by the 2024 Universitas Negeri Malang PNBP research grant (Contract number: 4.4.602/ UN32.14.1/LT/2024).

Conflict of interest

There was no conflict of interest among all authors in this study.

References

[1] Dekker E, Tanis PJ, Vleugels JL, Kasi PM, Wallace MB. Colorectal cancer. Lancet. 2019; 394(10207):1467-1480. doi: 10.1016/S0140-6736(19)32319-0

[2] Xi Y, Xu P. Global colorectal cancer burden in 2020 and projections to 2040. Transl Oncol. 2021; 14(10):101174. doi: 10.1016/j.tranon.2021.101174

[3] Li J, Ma X, Chakravarti D, Shalapour S, DePinho RA. Genetic and biological hallmarks of colorectal cancer. Genes Dev. 2021; 35(11-12):787-820. doi: 10.1101/gad.348226.120

[4] Kuipers EJ, Grady WM, Lieberman D, et al. Colorectal cancer. Nat Rev Dis Primers. 2015; 1:15065. doi: 10.1038/nrdp.2015.65

[5] Thanikachalam K, Khan G. Colorectal cancer and nutrition. Nutrients. 2019; 11(1):164. doi: 10.3390/nu11010164

[6] Brody H. Colorectal cancer. Nature. 2015; 521(7551):S1. doi: 10.1038/521S1a

[7] Lengauer C, Kinzler KW, Vogelstein B. Genetic instability in colorectal cancers. Nature. 1997; 386(6625):623-627. doi: 10.1038/386623a0

[8] Patel SG, Ahnen DJ. Colorectal cancer in the young. Curr Gastroenterol Rep. 2018; 20(4):15. doi: 10.1007/s11894-018-0618-9

[9] Zhou E, Rifkin S. Colorectal cancer and diet: Risk versus prevention, is diet an intervention? Gastroenterol Clin North Am. 2021; 50(1):101-111. doi: 10.1016/j.gtc.2020.10.012

[10] Valderrama-Treviño AI, Barrera-Mera B, Ceballos-Villalva JC, Montalvo-Javé EE. Hepatic metastasis from colorectal cancer. Euroasian J Hepatogastroenterol. 2017; 7(2):166-175. doi: 10.5005/jp-journals-10018-1241

[11] Martin J, Petrillo A, Smyth EC, et al. Colorectal liver metastases: Current management and future perspectives. World J Clin Oncol. 2020; 11(10):761-808. doi: 10.5306/wjco.v11.i10.761

[12] Zhou H, Liu Z, Wang Y, et al. Colorectal liver metastasis: Molecular mechanism and interventional therapy. Sig Transduct Target Ther. 2022; 7(1):1-25. doi: 10.1038/s41392-022-00922-2

[13] Weitz J, Koch M, Debus J, Höhler T, Galle PR, Büchler MW. Colorectal cancer. Lancet. 2005; 365(9454):153-165. doi: 10.1016/S0140-6736(05)17706-X

[14] Heinz MC, Peters NA, Oost KC, et al. Liver colonization by colorectal cancer metastases requires YAP-controlled plasticity at the micrometastatic stage. Cancer Res. 2022; 82(10):1953-1968. doi: 10.1158/0008-5472.CAN-21-0933

[15] Fu Z, Berhane F, Fite A, Seyoum B, Abou-Samra AB, Zhang R. Elevated circulating lipasin/betatrophin in human type 2 diabetes and obesity. Sci Rep. 2015; 4(1):5013. doi: 10.1038/srep05013

[16] Zhang R. The ANGPTL3-4-8 model, a molecular mechanism for triglyceride trafficking. Open Biol. 2016; 6(4):150272. doi: 10.1098/rsob.150272

[17] Gao T, Jin K, Chen P, et al. Circulating betatrophin correlates with triglycerides and postprandial glucose among different glucose tolerance statuses-a case-control study. PLoS One. 2015; 10(8):e0133640. doi: 10.1371/journal.pone.0133640

[18] Tseng YH, Yeh YH, Chen WJ, Lin KH. Emerging regulation and function of betatrophin. IJMS. 2014; 15(12): 23640-23657. doi: 10.3390/ijms151223640

[19] Sertogullarindan B, Komuroglu AU, Ucler R, Gunbatar H, Sunnetcioglu A, Cokluk E. Betatrophin association with serum triglyceride levels in obstructive sleep apnea patients. Ann Thorac Med. 2019; 14(1):63-68. doi: 10.4103/atm.ATM_52_18

[20] Xu F, Chen Y, Wang N, Sun K. Bacteria-derived recombinant human ANGPTL8/betatrophin significantly increases the level of triglyceride. Protein J. 2019; 38(4):472-478. doi: 10.1007/s10930-019-09825-8

[21] Hassan AB, Salih SF, Hassan II, et al. Circulating betatrophin in relation to metabolic, inflammatory parameters, and oxidative stress in patients with type 2 diabetes mellitus. Diabetes Metab Syndr. 2019; 13(1):458-463. doi: 10.1016/j.dsx.2018.11.016

[22] Wang Y, Quagliarini F, Gusarova V, et al. Mice lacking ANGPTL8 (Betatrophin) manifest disrupted triglyceride metabolism without impaired glucose homeostasis. Proc Natl Acad Sci USA. 2013; 110(40):16109-16114. doi: 10.1073/pnas.1315292110

[23] Fenzl A, Itariu BK, Kosi L, et al. Circulating betatrophin correlates with atherogenic lipid profiles but not with glucose and insulin levels in insulin-resistant individuals. Diabetologia. 2014; 57(6):1204-1208. doi: 10.1007/s00125-014-3208-x

[24] Hu H, Sun W, Yu S, et al. Increased circulating levels of betatrophin in newly diagnosed type 2 diabetic patients. Diabetes Care. 2014; 37:2718-2722. doi: 10.2337/dc14-0602

[25] Zhang R, Abou-Samra AB. A dual role of lipasin (betatrophin) in lipid metabolism and glucose homeostasis: consensus and controversy. Cardiovasc Diabetol. 2014; 13(1):133. doi: 10.1186/s12933-014-0133-8

[26] Abu-Farha M, Cherian P, Qaddoumi MG, et al. Increased plasma and adipose tissue levels of ANGPTL8/Betatrophin and ANGPTL4 in people with hypertension. Lipids Health Dis. 2018; 17(1):35. doi: 10.1186/s12944-018-0681-0

[27] Liu J, Yagi K, Nohara A, et al. High frequency of type 2 diabetes and impaired glucose tolerance in Japanese subjects with the angiopoietin-like protein 8 R59W variant. J Clin Lipidol. 2018; 12(2):331-337. doi: 10.1016/j.jacl.2017.12.011

[28] Gómez-Ambrosi J, Pascual-Corrales E, Catalán V, et al. Altered concentrations in dyslipidemia evidence a role for ANGPTL8/betatrophin in lipid metabolism in humans. J Clin Endocrinol Metab. 2016; 101(10):3803-3811. doi: 10.1210/jc.2016-2084

[29] Ye J, Qin Y, Wang D, Yang L, Yuan G. The relationship between circulating ANGPTL8/betatrophin concentrations and adult obesity: A meta-analysis. Dis Markers. 2019; 2019:5096860. doi: 10.1155/2019/5096860

[30] Wu S, Gao H, Ma Y, Fu L, Zhang C, Luo X. Characterisation of betatrophin concentrations in childhood and adolescent obesity and insulin resistance. Pediatr Diabetes. 2016; 17(1):53-60. doi: 10.1111/pedi.12233

[31] Guo K, Lu J, Yu H, et al. Serum betatrophin concentrations are significantly increased in overweight but not in obese or type 2 diabetic individuals. Obesity (Silver Spring). 2015; 23(4):793-797. doi: 10.1002/oby.21038

[32] Wang H, Lai Y, Han C, et al. The effects of serum ANGPTL8/ betatrophin on the risk of developing the metabolic syndrome - a prospective study. Sci Rep. 2016; 6(1):28431. doi: 10.1038/srep28431

[33] Chen CC, Susanto H, Chuang WH, Liu TY, Wang CH. Higher serum betatrophin level in type 2 diabetes subjects is associated with urinary albumin excretion and renal function. Cardiovasc Diabetol. 2016; 15:3. doi: 10.1186/s12933-015-0326-9

[34] Pazgan-Simon M, Zuwala-Jagiello J, Menzyk T, et al. Serum betatrophin and irisin levels in hepatocellular carcinoma. J Physiol Pharmacol. 2020; 71(1): 113-123. doi: 10.26402/jpp.2020.1.11

[35] Hong BS, Liu J, Zheng J, et al. Angiopoietin-like protein 8/betatrophin correlates with hepatocellular lipid content independent of insulin resistance in non-alcoholic fatty liver disease patients. J Diabetes Investig. 2018; 9(4):952-958. doi: 10.1111/jdi.12792

[36] Xu F, Tian D, Shi X, Sun K, Chen Y. Analysis of the expression and prognostic potential of a novel metabolic regulator ANGPTL8/betatrophin in human cancers. Pathol Oncol Res. 2021; 27:1609914. doi: 10.3389/pore.2021.1609914

[37] Taherkhani F, Hosseini KM, Zebardast S, Chegini KG, Gheibi N. Anti proliferative and apoptotic effects on pancreatic cancer cell lines indicate new roles for ANGPTL8 (Betatrophin). Genet Mol Biol. 2020; 43(3):e20190196. doi: 10.1590/1678-4685-gmb-2019-0196

[38] Monzavi N, Zargar SJ, Gheibi N, Azad M, Rahmani B. Angiopoietin-like protein 8 (betatrophin) may inhibit hepatocellular carcinoma through suppressing of the Wnt signaling pathway. Iran J Basic Med Sci. 2019; 22(10):1166. doi: 10.22038/ijbms.2019.36612.8764

[39] Gao Y, Yuan Y, Wen S, et al. Dual role of ANGPTL8 in promoting tumor cell proliferation and immune escape during hepatocarcinogenesis. Oncogenesis. 2023; 12(1):26. doi: 10.1038/s41389-023-00473-3

[40] Navaeian M, Ahmadpour-Yazdi H, Asadian S, Gheibi N. The effect of ANGPTL8 protein on proliferation and apoptosis in HepG2 hepatocellular carcinoma cell line. Gene Reports. 2021; 25:101306. doi: 10.1016/j.genrep.2021.101306

[41] Huot JR, Novinger LJ, Pin F, et al. Formation of colorectal liver metastases induces musculoskeletal and metabolic abnormalities consistent with exacerbated cachexia. JCI Insight. 2020; 5(9):e136687. doi: 10.1172/jci.insight.136687

[42] Shiono M, Huang K, Downey RJ, et al. An analysis of the relationship between metastases and cachexia in lung cancer patients. Cancer Med. 2016; 5(9):2641-2648. doi: 10.1002/cam4.841

[43] Yi M, Chen RP, Yang R, Guo XF, Zhang JC, Chen H. Betatrophin acts as a diagnostic biomarker in type 2 diabetes mellitus and is negatively associated with HDL-cholesterol. Int J Endocrinol. 2015; 2015:479157. doi: 10.1155/2015/479157

[44] Li S, Liu D, Li L, et al. Circulating betatrophin in patients with type 2 diabetes: A meta-analysis. J Diabetes Res. 2016; 2016:6194750. doi: 10.1155/2016/6194750

[45] Abu-Farha M, Sriraman D, Cherian P, et al. Circulating ANGPTL8/betatrophin is increased in obesity and reduced after exercise training. PLOS One. 2016; 11(1):e0147367. doi: 10.1371/journal.pone.0147367

[46] Zheng J, Liu J, Hong BS, Ke W, Huang M, Li Y. Circulating betatrophin/ANGPTL8 levels correlate with body fat distribution in individuals with normal glucose tolerance but not those with glucose disorders. BMC Endocr Disord. 2020; 20:51. doi: 10.1186/s12902-020-0531-8

[47] Susanto H, Liu TY, Chen CC, Purnomo JD, Chen SF, Wang CH. Increased serum levels of betatrophin in pancreatic cancer-associated diabetes. Oncotarget. 2106; 7(27):42330-42339. doi: 10.18632/oncotarget.9815

[48] Carbone C, Piro G, Merz V, et al. Angiopoietin-like proteins in angiogenesis, inflammation and cancer. Int J Mol Sci. 2018; 19(2):431. doi: 10.3390/ijms19020431

[49] Susanto H, Firdaus SR, Sholeh M, et al. Moringa oleifera leaf powder - silver nanoparticles (MOLP-AgNPs) efficiently inhibit metastasis and proliferative signaling in HT-29 human colorectal cancer cells. J Agric Food Res. 2024; 16:101149. doi: 10.1016/j.jafr.2024.101149

[50] Abu-Farha M, Al-Khairi I, Cherian P, et al. Increased ANGPTL3, 4 and ANGPTL8/betatrophin expression levels in obesity and T2D. Lipids Health Dis. 2016; 15(1):181. doi: 10.1186/s12944-016-0337-x

[51] Luo M, Peng D. ANGPTL8: An Important regulator in metabolic disorders. Front Endocrinol (Lausanne). 2018; 9:169. doi: 10.3389/fendo.2018.00169

[52] Guo Q, Cao S, Wang X. Betatrophin and insulin resistance. Metabolites. 2022; 12(10):925. doi: 10.3390/metabo12100925

[53] Maurer L, Brachs S, Decker AM, et al. Weight loss partially restores glucose-driven betatrophin response in humans. J Clin Endocrinol Metab. 2016; 101(11):4014-4020. doi: 10.1210/jc.2016-1788

[54] Susanto H, Taufiq A, Listyorini D, Handaya AY, Pertiwi MP. Protein-based biomaterial marker in metabolic syndrome and colorectal cancer: A preliminary clinical study of betatrophin expression in Javanese Ethnic. IOP Conf Ser Mater Sci Eng. 2019; 515:012054. doi: 10.1088/1757-899X/515/1/012054

[55] Susanto H. Betatrophin: The long-term exploration of potential biomarker for metabolic syndrome and cancer-related metabolic perturbation. AIP Conf Proc. 2025; 3186(1):020002. doi: 10.1063/5.0234671