Expression and clinical significance of osteocalcin in endometrial carcinoma

Background: Steocalcin (OCN) is a bone tissue-specific protein overexpressed in several human cancers. However, the physiological roles of OCN in endometrial cancer (EC) are unclear. This study aimed to investigate its prognostic impact on OCN expression in EC patients and their associations with clinicopathologic features. Methods: Between July 2010 and December 2013 at the Department of Obstetrics and Gynecology, Shanghai Pudong Hospital, we performed immunohistochemistry to detect OCN expression in 134 EC patients and 35 normal endometria (non-EC) specimens and analyze its correlation with clinicopathologic features. The enzyme-linked immunosorbent assay (ELISA) analyzed the OCN expression in serum. OCN expression was also detected in fresh tissues using real-time polymerase chain reaction (PCR) and western blotting. Results: OCN mRNA and protein expressions were significantly lower in EC tissues than adjacent normal tissues (p < 0.001). Low OCN expression was significantly associated with the International Federation of Gynecology and Obstetrics (FIGO) stage, lymph node metastasis, and depth of myometrial invasion (p < 0.01). Kaplan-Meier analysis exposed a significant difference in overall survival (OS) and disease-free survival (DFS) between EC patients with low and high OCN expression levels (log-rank, both p < 0.001). Moreover, multivariate Cox regression analysis showed that OCN expression was an independent predictive factor for overall survival (OS) (p = 0.043) and disease-free survival (DFS) (p = 0.027) in EC patients. Spearman’s rank correlation analysis stated a positive correlation between serum OCN level and adiponectin (r = 0.455, p < 0.001) and a negative correlation between serum OCN level and leptin (r = −0.307, p < 0.001). Conclusions: The results speculate that the low OCN expression is associated with the progression and recurrence of endometrial cancer.

Endometrial cancer; Osteocalcin; Prognosis; Immunohistochemistry

[1] Siegel RL, Miller KD, Jemal A. Cancer statistics, 2017. CA: A Cancer Journal for Clinicians. 2017; 67: 7–30.

[2] Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, et al. Cancer statistics in China, 2015. CA: A Cancer Journal for Clinicians. 2016; 66: 115–132.

[3] Cowan M, Strauss JB, Barber EL, Matei D. Updates on adjuvant chemotherapy and radiation therapy for endometrial cancer. Current Opinion in Obstetrics & Gynecology. 2019; 31: 31–37.

[4] Rodriguez AC, Blanchard Z, Maurer KA, Gertz J. Estrogen signaling in endometrial cancer: a key oncogenic pathway with several open questions. Hormones & Cancer. 2019; 10: 51–63.

[5] Hernandez AV, Pasupuleti V, Benites-Zapata VA, Thota P, Deshpande A, Perez-Lopez FR. Insulin resistance and endometrial cancer risk: a systematic review and meta-analysis. European Journal of Cancer. 2015; 51: 2747–2758.

[6] Onstad MA, Schmandt RE, Lu KH. Addressing the role of obesity in endometrial cancer risk, prevention, and treatment. Journal of Clinical Oncology. 2016; 34: 4225–4230.

[7] Tumminia A, Vinciguerra F, Parisi M, Graziano M, Sciacca L, Baratta R, et al. Adipose tissue, obesity and adiponectin: role in endocrine cancer risk. International Journal of Molecular Sciences. 2019; 20: 2863.

[8] Busch EL, Crous-Bou M, Prescott J, Downing MJ, Rosner BA, Mutter GL, et al. Adiponectin, leptin, and insulin-pathway receptors as endometrial cancer subtyping markers. Hormones and Cancer. 2018; 9: 33–39.

[9] Ellis PE, Barron GA, Bermano G. Adipocytokines and their relationship to endometrial cancer risk: a systematic review and meta-analysis. Gynecologic Oncology. 2020; 158: 507–516.

[10] Ma Y, Liu Z, Zhang Y, Lu B. Serum leptin, adiponectin and endometrial cancer risk in Chinese women. Journal of Gynecologic Oncology. 2013; 24: 336–341.

[11] Gong T, Wu Q, Wang Y, Ma X. Circulating adiponectin, leptin and adiponectin-leptin ratio and endometrial cancer risk: evidence from a meta-analysis of epidemiologic studies. International Journal of Cancer. 2015; 137: 1967–1978.

[12] Al-Suhaimi EA, Al-Jafary MA. Endocrine roles of vitamin K-dependent- osteocalcin in the relation between bone metabolism and metabolic disorders. Reviews in Endocrine and Metabolic Disorders. 2020; 21: 117–125.

[13] Franzén A, Hultenby K, Reinholt FP, Önnerfjord P, Heinegård D. Altered osteoclast development and function in osteopontin deficient mice. Journal of Orthopaedic Research. 2008; 26: 721–728.

[14] Razny U, Fedak D, Kiec-Wilk B, Goralska J, Gruca A, Zdzienicka A, et al. Carboxylated and undercarboxylated osteocalcin in metabolic complications of human obesity and prediabetes. Diabetes/Metabolism Research and Reviews. 2017; 33: e2862.

[15] Mizokami A, Kawakubo-Yasukochi T, Hirata M. Osteocalcin and its endocrine functions. Biochemical Pharmacology. 2017; 132: 1–8.

[16] Nimptsch K, Rohrmann S, Nieters A, Linseisen J. Serum undercarboxylated osteocalcin as biomarker of vitamin k intake and risk of prostate cancer: a nested case-control study in the heidelberg cohort of the european prospective investigation into cancer and nutrition. Cancer Epidemiology, Biomarkers & Prevention. 2009; 18: 49–56.

[17] Delaka A, Mourmouras N, Zervou-Valvi F, Stravodimos K, Anastasiou I, Delakas D. Predictive value of the bone markers osteocalcin and C-terminal telopeptide for jaw osteonecrosis in high-risk prostate cancer patients on bisphosphonate therapy: a prospective study. Archivos Espanoles De Urologia. 2019; 72: 948–954.

[18] Lee K, Lee KJ, Kim T, Hutomo F, Sun HJ, Cheon GJ, et al. Circulating osteocalcin-positive cells as a novel diagnostic biomarker for bone metastasis in breast cancer patients. Journal of Bone and Mineral Research. 2020; 35: 1838–1849.

[19] Benedet JL, Bender H, Jones H III, Ngan HY, Pecorelli S. FIGO staging classifications and clinical practice guidelines in the management of gynecologic cancers. FIGO committee on gynecologic oncology. International Journal of Gynaecology and Obstetrics. 2000; 70: 209–262.

[20] Penzel R, Hoegel J, Schmitz W, Blaeker H, Morresi-Hauf A, Aulmann S, et al. Clusters of chromosomal imbalances in thymic epithelial tumours are associated with the who classification and the staging system according to Masaoka. International Journal of Cancer. 2003; 105: 494–498.

[21] Agustina H, Asyifa I, Aziz A, Hernowo BS. The role of osteocalcin and alkaline phosphatase immunohistochemistry in osteosarcoma diagnosis. Pathology Research International. 2018; 2018: 1–5.

[22] Amaral CB, Leite JDS, Fonseca ABM, Ferreira AMR. Vimentin, osteocalcin and osteonectin expression in canine primary bone tumors: diagnostic and prognostic implications. Molecular Biology Reports. 2018; 45: 1289–1296.

[23] Salem AM, Zohny SF, Abd El-Wahab MM, Hamdy R. Predictive value of osteocalcin and beta-CrosLaps in metastatic breast cancer. Clinical Biochemistry. 2007; 40: 1201–1208.

[24] Wang J, Xu J, Wang B, Shu FR, Chen K, Mi MT. Equol promotes rat osteoblast proliferation and differentiation through activating estrogen receptor. Genetics and Molecular Research. 2014; 13: 5055–5563.

[25] Liu Y, Huang L, Lu Y, Xi XE, Huang XL, Lu Q, et al. Relationships between the osteocalcin gene polymorphisms, serum osteocalcin levels, and hepatitis B virus-related hepatocellular carcinoma in a Chinese population. PLOS ONE. 2015; 10: e0116479.

[26] Lin X, Brennan-Speranza TC, Levinger I, Yeap BB. Undercarboxylated osteocalcin: experimental and human evidence for a role in glucose homeostasis and muscle regulation of insulin sensitivity. Nutrients. 2018; 10: 847.

[27] Hayashi Y, Kawakubo-Yasukochi T, Mizokami A, Takeuchi H, Nakamura S, Hirata M. Differential roles of carboxylated and uncarboxylated osteocalcin in prostate cancer growth. Journal of Cancer. 2016; 7: 1605–1609.

[28] Hayashi Y, Kawakubo-Yasukochi T, Mizokami A, Hazekawa M, Yakura T, Naito M, et al. Uncarboxylated osteocalcin induces antitumor immunity against mouse melanoma cell growth. Journal of Cancer. 2017; 8: 2478–2486.

[29] Gardner TA, Lee SJ, Lee SD, Li X, Shirakawa T, Kwon DD, et al. Differential expression of osteocalcin during the metastatic progression of prostate cancer. Oncology Reports. 2009; 21: 903–908.

[30] Wang Z, Gao S, Sun C, Li J, Gao W, Yu L. Clinical significance of serum adiponectin and visfatin levels in endometrial cancer. International Journal of Gynecology & Obstetrics. 2019; 145: 34–39.

[31] Tan W, Wang L, Ma Q, Qi M, Lu N, Zhang L, et al. Adiponectin as a potential tumor suppressor inhibiting epithelial-to-mesenchymal transition but frequently silenced in prostate cancer by promoter methylation. The Prostate. 2015; 75: 1197–1205.

[32] Yadav A, Kataria MA, Saini V, Yadav A. Role of leptin and adiponectin in insulin resistance. Clinica Chimica Acta. 2013; 417: 80–84.

[33] Iki M, Tamaki J, Fujita Y, Kouda K, Yura A, Kadowaki E, et al. Serum undercarboxylated osteocalcin levels are inversely associated with glycemic status and insulin resistance in an elderly Japanese male population: fujiwara-kyo osteoporosis risk in men (FORMEN) study. Osteoporosis International. 2012; 23: 761–770.

[34] Díaz-López A, Bulló M, Juanola-Falgarona M, Martínez-González MA, Estruch R, Covas M, et al. Reduced serum concentrations of carboxylated and undercarboxylated osteocalcin are associated with risk of developing type 2 diabetes mellitus in a high cardiovascular risk population: a nested case-control study. The Journal of Clinical Endocrinology & Metabolism. 2013; 98: 4524–4531.

[35] Zhang X, Wang Y, Ma L, Liu Z, Ye F, Yang J. Uncarboxylated osteocalcin ameliorates hepatic glucose and lipid metabolism in KKAy mice via activating insulin signaling pathway. Acta Pharmacologica Sinica. 2020; 41: 383–393.

[36] Guedes JAC, Esteves JV, Morais MR, Zorn TM, Furuya DT. Osteocalcin improves insulin resistance and inflammation in obese mice: participation of white adipose tissue and bone. Bone. 2018; 115: 68–82.

[37] Bilotta FL, Arcidiacono B, Messineo S, Greco M, Chiefari E, Britti D, et al. Insulin and osteocalcin: further evidence for a mutual cross-talk. Endocrine. 2018; 59: 622–632.

[38] Coskun G, Sencar L, Tuli A, Saker D, Alparslan MM, Polat S. Effects of osteocalcin on synthesis of testosterone and INSL3 during adult leydig cell differentiation. International Journal of Endocrinology. 2019; 2019: 1–18.

[39] Yeap BB, Alfonso H, Chubb SA, Gauci R, Byrnes E, Beilby JP, et al. Higher serum undercarboxylated osteocalcin and other bone turnover markers are associated with reduced diabetes risk and lower estradiol concentrations in older men. The Journal of Clinical Endocrinology and Metabolism. 2015; 100: 63–71.