Endocrinol Metab.  2021 Feb;36(1):96-105. 10.3803/EnM.2020.819.

Lactate Dehydrogenase A as a Potential New Biomarker for Thyroid Cancer

• Ban EJ
• Kim D
• Kim JK
• Kang SW
• Lee J
• Jeong JJ
• Nam KH
• Chung WY
• Kim K

• KMID: 2513293
• DOI: http://doi.org/10.3803/EnM.2020.819

Abstract

Background
Several cancers show increased levels of lactate dehydrogenase A (LDHA), which are associated with cancer progression. However, it remains unclear whether LDHA levels are associated with papillary thyroid cancer (PTC) aggressiveness or with the presence of the PTC prognostic marker, the BRAFV^600E mutation. This study aimed to evaluate the potential of LDHA as a PTC prognostic marker.
Methods
LDHA expression was examined in 83 PTC tissue specimens by immunohistochemistry. Human thyroid cell lines were genetically manipulated to overexpress BRAFV^600E or were treated with a BRAF-specific short hairpin RNA (shBRAF), whose effects on LDHA expression were evaluated by Western blotting. Data from 465 PTC patients were obtained from The Cancer Genome Atlas (TCGA) database and analyzed to validate the in vitro results.
Results
LDHA was aberrantly overexpressed in PTC. Intense immunostaining for LDHA was observed in PTC specimens carrying mutated BRAF, whereas the intensity was less in wild-type BRAF samples. Overexpression of BRAFV^600E resulted in LDHA upregulation, whereas treatment with shBRAF downregulated LDHA in human thyroid cell lines. Furthermore, LDHA mRNA expression was significantly elevated and associated with BRAFV^600E expression in thyroid cancer tissues from TCGA database. Additionally, LDHA overexpression was found to be correlated with aggressive clinical features of PTC, such as lymph node metastases and advanced tumor stages.
Conclusion
LDHA overexpression is associated with the BRAFV^600E mutation and an aggressive PTC behavior. Therefore, LDHA may serve as a biomarker and therapeutic target in PTC.

Keyword

Biomarkers; BRAF; Lactate dehydrogenase A; Thyroid cancer, papillary; Prognosis

Figure

• 

  Fig. 1 Lactate dehydrogenase A (LDHA) expression in normal and tumor tissues analyzed by immunohistochemistry. (A) Normal thyroid tissue; (B) weakly, (C) intermediately, and (D) strongly stained papillary thyroid carcinoma tissues (×400).

• 

  Fig. 2 Protein expression in Nthy-ori 3-1 cells stably expressing the wild-type (WT) BRAF or BRAF with the V600E mutation. (A) Photomicrographs of BRAFV600E- and WT BRAF-overexpressing cells. (B) BRAF, extracellular signal-regulated kinase (ERK), phosphorylated ERK (pERK), lactate dehydrogenase A (LDHA), and β-actin levels in BRAFV600E- and WT BRAF-overexpressing cells. The blot is representative of three independent experiments.

• 

  Fig. 3 Lactate dehydrogenase A (LDHA) expression in human thyroid cell lines after BRAF-specific short hairpin RNA (shBRAF) treatment. (A) SNU-790 and (B) 8505c cells. Each blot is representative of three independent experiments.

• 

  Fig. 4 mRNA expression of glycolytic enzyme-encoding genes based on The Cancer Genome Atlas thyroid cancer database. (A) Glucose transporter 1 (GLUT1), (B) pyruvate kinase isoform M2 (PKM2), and (C) lactate dehydrogenase A (LDHA) expression levels in 465 papillary thyroid carcinoma and 55 normal thyroid tissues. Data are presented as the individual value, median, and interquartile range. RSEM, RNA-Seq by expectation-maximization.

• 

  Fig. 5 Lactate dehydrogenase A (LDHA) gene expression levels according to the BRAFV600E mutational status or papillary thyroid cancer (PTC) subtype, based on The Cancer Genome Atlas (TCGA) thyroid cancer database. (A) LDHA mRNA expression levels in BRAF wild-type (WT) and V600E mutant PTC. (B) LDHA mRNA expression levels according to the subtype of PTC. Data are presented as the individual value, median, and interquartile range. RSEM, RNA-Seq by expectation-maximization. aClassic, classical/usual; bFV, follicular (≥99% follicular patterned); cTV, tall-cell (≥50% tall cell features); dOther, uncommon PTC variants.

Reference

1. Feng Y, Xiong Y, Qiao T, Li X, Jia L, Han Y. Lactate dehydrogenase A: a key player in carcinogenesis and potential target in cancer therapy. Cancer Med. 2018; 7:6124–36.

2. Woodford MR, Chen VZ, Backe SJ, Bratslavsky G, Mollapour M. Structural and functional regulation of lactate dehydrogenase-A in cancer. Future Med Chem. 2020; 12:439–55.

3. Yao F, Zhao T, Zhong C, Zhu J, Zhao H. LDHA is necessary for the tumorigenicity of esophageal squamous cell carcinoma. Tumour Biol. 2013; 34:25–31.

4. Girgis H, Masui O, White NM, Scorilas A, Rotondo F, Seivwright A, et al. Lactate dehydrogenase A is a potential prognostic marker in clear cell renal cell carcinoma. Mol Cancer. 2014; 13:101.

5. Sun X, Sun Z, Zhu Z, Guan H, Zhang J, Zhang Y, et al. Clinicopathological significance and prognostic value of lactate dehydrogenase A expression in gastric cancer patients. PLoS One. 2014; 9:e91068.

6. Lim H, Devesa SS, Sosa JA, Check D, Kitahara CM. Trends in thyroid cancer incidence and mortality in the United States, 1974–2013. JAMA. 2017; 317:1338–48.

7. Kim WB, Jeon MJ, Kim WG, Kim TY, Shong YK. Unmet clinical needs in the treatment of patients with thyroid cancer. Endocrinol Metab (Seoul). 2020; 35:14–25.

8. Wiltshire JJ, Drake TM, Uttley L, Balasubramanian SP. Systematic review of trends in the incidence rates of thyroid cancer. Thyroid. 2016; 26:1541–52.

9. Chung JH. BRAF and TERT promoter mutations: clinical application in thyroid cancer. Endocr J. 2020; 67:577–84.

10. Kim TH, Park YJ, Lim JA, Ahn HY, Lee EK, Lee YJ, et al. The association of the BRAF(V600E) mutation with prognostic factors and poor clinical outcome in papillary thyroid cancer: a meta-analysis. Cancer. 2012; 118:1764–73.

11. Song YS, Lim JA, Park YJ. Mutation profile of well-differentiated thyroid cancer in Asians. Endocrinol Metab (Seoul). 2015; 30:252–62.

12. Wen SS, Zhang TT, Xue DX, Wu WL, Wang YL, Wang Y, et al. Metabolic reprogramming and its clinical application in thyroid cancer. Oncol Lett. 2019; 18:1579–84.

13. Kachel P, Trojanowicz B, Sekulla C, Prenzel H, Dralle H, Hoang-Vu C. Phosphorylation of pyruvate kinase M2 and lactate dehydrogenase A by fibroblast growth factor receptor 1 in benign and malignant thyroid tissue. BMC Cancer. 2015; 15:140.

14. Ciampi R, Vivaldi A, Romei C, Del Guerra A, Salvadori P, Cosci B, et al. Expression analysis of facilitative glucose transporters (GLUTs) in human thyroid carcinoma cell lines and primary tumors. Mol Cell Endocrinol. 2008; 291:57–62.

15. Gao Y, Yang F, Yang XA, Zhang L, Yu H, Cheng X, et al. Mitochondrial metabolism is inhibited by the HIF1α-MYC-PGC-1β axis in BRAF V600E thyroid cancer. FEBS J. 2019; 286:1420–36.

16. Cancer Genome Atlas Research Network. Integrated genomic characterization of papillary thyroid carcinoma. Cell. 2014; 159:676–90.

17. Cancer Genome Atlas Research Network. Weinstein JN, Collisson EA, Mills GB, Shaw KR, Ozenberger BA, et al. The cancer genome atlas pan-cancer analysis project. Nat Genet. 2013; 45:1113–20.

18. Kim YH, Choi YW, Han JH, Lee J, Soh EY, Park SH, et al. TSH signaling overcomes B-RafV600E-induced senescence in papillary thyroid carcinogenesis through regulation of DUSP6. Neoplasia. 2014; 16:1107–20.

19. Ko H, Kim S, Jin CH, Lee E, Ham S, Yook JI, et al. Protein kinase casein kinase 2-mediated upregulation of N-cadherin confers anoikis resistance on esophageal carcinoma cells. Mol Cancer Res. 2012; 10:1032–8.

20. Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011; 144:646–74.

21. Iniguez-Ariza NM, Brito JP. Management of low-risk papillary thyroid cancer. Endocrinol Metab (Seoul). 2018; 33:185–94.

22. Jeon MJ, Kim WG, Kim TH, Kim HK, Kim BH, Yi HS, et al. Disease-specific mortality of differentiated thyroid cancer patients in Korea: a multicenter cohort study. Endocrinol Metab (Seoul). 2017; 32:434–41.

23. Grabellus F, Worm K, Schmid KW, Sheu SY. The BRAF V600E mutation in papillary thyroid carcinoma is associated with glucose transporter 1 overexpression. Thyroid. 2012; 22:377–82.

24. Miao P, Sheng S, Sun X, Liu J, Huang G. Lactate dehydrogenase A in cancer: a promising target for diagnosis and therapy. IUBMB Life. 2013; 65:904–10.

25. Kim BA, Jee HG, Yi JW, Kim SJ, Chai YJ, Choi JY, et al. Expression profiling of a human thyroid cell line stably expressing the BRAFV600E mutation. Cancer Genomics Proteomics. 2017; 14:53–67.

26. Feng C, Gao Y, Wang C, Yu X, Zhang W, Guan H, et al. Aberrant overexpression of pyruvate kinase M2 is associated with aggressive tumor features and the BRAF mutation in papillary thyroid cancer. J Clin Endocrinol Metab. 2013; 98:E1524–33.

27. Feine U, Lietzenmayer R, Hanke JP, Held J, Wohrle H, Muller-Schauenburg W. Fluorine-18-FDG and iodine-131-iodide uptake in thyroid cancer. J Nucl Med. 1996; 37:1468–72.

28. Guo J, Higashi K, Yokota H, Nagao Y, Ueda Y, Kodama Y, et al. In vitro proton magnetic resonance spectroscopic lactate and choline measurements, 18F-FDG uptake, and prognosis in patients with lung adenocarcinoma. J Nucl Med. 2004; 45:1334–9.

29. Barollo S, Pennelli G, Vianello F, Watutantrige Fernando S, Negro I, Merante Boschin I, et al. BRAF in primary and recurrent papillary thyroid cancers: the relationship with (131)I and 2-[(18)F]fluoro-2-deoxy-D-glucose uptake ability. Eur J Endocrinol. 2010; 163:659–63.

30. Liu R, Bishop J, Zhu G, Zhang T, Ladenson PW, Xing M. Mortality risk stratification by combining BRAF V600E and TERT promoter mutations in papillary thyroid cancer: genetic duet of BRAF and TERT promoter mutations in thyroid cancer mortality. JAMA Oncol. 2017; 3:202–8.

31. Xing M. Genetic-guided risk assessment and management of thyroid cancer. Endocrinol Metab Clin North Am. 2019; 48:109–24.