Effects of GPR81 silencing combined with cisplatin stimulation on biological function in hypopharyngeal squamous cell carcinoma

Jia,Qiaojing; Xu,Ou; Wang,Jianxing; Dong,Jinhui; Ren,Xiumin; Jia,Xiaofang; Shan,Chunguang

doi:10.3892/mmr.2020.11255

Effects of GPR81 silencing combined with cisplatin stimulation on biological function in hypopharyngeal squamous cell carcinoma

Authors:
- Qiaojing Jia
- Ou Xu
- Jianxing Wang
- Jinhui Dong
- Xiumin Ren
- Xiaofang Jia
- Chunguang Shan
View Affiliations

Affiliations: E.N.T. Department, The Second Hospital of Hebei Medical University, Shijiazhuang, Hebei 050000, P.R. China
Published online on: June 18, 2020 https://doi.org/10.3892/mmr.2020.11255
Pages: 1727-1736
Copyright: © Jia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hypopharyngeal squamous cell carcinoma (HSCC) is a malignant tumor found in the head and neck region. Lactate receptor 1, also known as G protein‑coupled receptor81 (GPR81), has been reported to play a vital role in cancer growth and metabolism. However, the function of GPR81 in HSCC remains largely unknown. The present study investigated the effect of GPR81 on cell survival and GPR81‑mediated energy metabolism under cisplatin treatment in HSCC. GPR81 knockdown reduced the proliferation and invasion of the human HSCC cell line FaDu. Furthermore, GPR81 silencing combined with cisplatin treatment increased the expression of translocase of outer mitochondrial membrane 20 at the mRNA and protein levels (P<0.05). mRNA and protein expression of phosphofructokinase 1 in mRNA appeared to be downregulated in GPR81 knockdown FaDu cells treated with cisplatin, although this was not statistically significant. GPR81 silencing and cisplatin challenge showed no significant upregulation compared with the control, but significant downregulation in mRNA and protein levels compared with the shRNA‑scramble group. Apoptosis was measured by flow cytometry with annexin V and 7‑aminoactinomycin D. GPR81 silencing and cisplatin led to an increased apoptotic rate. Moreover, absence of GPR81 combined with cisplatin exposure increased caspase‑3 expression and decreased Bcl‑2 levels. The results of the present study suggested that GPR81 and cisplatin sensitivity played an important role in HSCC growth and metabolism.

View Figures

View References

1	Cohen N, Fedewa S and Chen AY: Epidemiology and demographics of the head and neck cancer population. Oral Maxillofac Surg Clin North Am. 30:381–395. 2018. View Article : Google Scholar : PubMed/NCBI
2	Bradley PJ: Epidemiologyz of hypopharyngeal cancer. Adv Otorhinolaryngol. 83:1–14. 2019.PubMed/NCBI
3	Znaor A, Brennan P, Gajalakshmi V, Mathew A, Shanta V, Varghese C and Boffetta P: Independent and combined effects of tobacco smoking, chewing and alcohol drinking on the risk of oral, pharyngeal and esophageal cancers in Indian men. Int J Cancer. 105:681–686. 2003. View Article : Google Scholar : PubMed/NCBI
4	Mehanna H, Beech T, Nicholson T, El-Hariry I, McConkey C, Paleri V and Roberts S: Prevalence of human papillomavirus in oropharyngeal and nonoropharyngeal head and neck cancer-systematic review and meta-analysis of trends by time and region. Head Neck. 35:747–755. 2013. View Article : Google Scholar : PubMed/NCBI
5	Newman JR, Connolly TM, Illing EA, Kilgore ML, Locher J and Carroll WR: Survival trends in hypopharyngeal cancer: A population-based review. Laryngoscope. 125:624–629. 2015. View Article : Google Scholar : PubMed/NCBI
6	American Cancer Society: Laryngeal and hypopharyngeal cancer. January 4–2015
7	Hanahan D and Weinberg RA: Hallmarks of cancer: The next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
8	Moreno-Sánchez R, Rodríguez-Enríquez S, Marín-Hernández A and Saavedra E: Energy metabolism in tumor cells. FEBS J. 274:1393–1418. 2007. View Article : Google Scholar : PubMed/NCBI
9	Fantin VR, St-Pierre J and Leder P: Attenuation of LDH-A expression uncovers a link between glycolysis, mitochondrial physiology, and tumor maintenance. Cancer Cell. 9:425–434. 2006. View Article : Google Scholar : PubMed/NCBI
10	Díaz-Muñiz I, Banavara DS, Budinich MF, Rankin SA, Dudley EG and Steele JL: Lactobacillus casei metabolic potential to utilize citrate as an energy source in ripening cheese: A bioinformatics approach. J Appl Microbiol. 101:872–882. 2006. View Article : Google Scholar : PubMed/NCBI
11	Brodsky AN, Odenwelder DC and Harcum SW: High extracellular lactate causes reductive carboxylation in breast tissue cell lines grown under normoxic conditions. PLoS One. 14:e02134192019. View Article : Google Scholar : PubMed/NCBI
12	Grasmann G, Smolle E, Olschewski H and Leithner K: Gluconeogenesis in cancer cells-Repurposing of a starvation-induced metabolic pathway? Biochim Biophys Acta Rev Cancer. 1872:24–36. 2019. View Article : Google Scholar : PubMed/NCBI
13	Liu C, Wu J, Zhu J, Kuei C, Yu J, Shelton J, Sutton SW, Li X, Yun SJ, Mirzadegan T, et al: Lactate inhibits lipolysis in fat cells through activation of an orphan G-protein-coupled receptor, GPR81. J Biol Chem. 284:2811–2822. 2009. View Article : Google Scholar : PubMed/NCBI
14	Rooney K and Trayhurn P: Lactate and the GPR81 receptor in metabolic regulation: Implications for adipose tissue function and fatty acid utilisation by muscle during exercise. Br J Nutr. 106:1310–1316. 2011. View Article : Google Scholar : PubMed/NCBI
15	Morland C, Lauritzen KH, Puchades M, Holm-Hansen S, Andersson K, Gjedde A, Attramadal H, Storm-Mathisen J and Bergersen LH: The lactate receptor, G-protein-coupled receptor 81/hydroxycarboxylic acid receptor 1: Expression and action in brain. J Neurosci Res. 93:1045–1055. 2015. View Article : Google Scholar : PubMed/NCBI
16	Wagner W, Kania KD, Blauz A and Ciszewski WM: The lactate receptor (HCAR1/GPR81) contributes to doxorubicin chemoresistance via ABCB1 transporter up-regulation in human cervical cancer HeLa cells. J Physiol Pharmacol. 68:555–564. 2017.PubMed/NCBI
17	Feng J, Yang H, Zhang Y, Wei H, Zhu Z, Zhu B, Yang M, Cao W, Wang L and Wu Z: Tumor cell-derived lactate induces TAZ-dependent upregulation of PD-L1 through GPR81 in human lung cancer cells. Oncogene. 36:5829–5839. 2017. View Article : Google Scholar : PubMed/NCBI
18	Roland CL, Arumugam T, Deng T, Liu SH, Philip B, Gomez S, Burns WR, Ramachandran V, Wang H, Cruz-Monserrate Z and Logsdon CD: Cell surface lactate receptor GPR81 is crucial for cancer cell survival. Cancer Res. 74:5301–5310. 2014. View Article : Google Scholar : PubMed/NCBI
19	Caslin HL, Abebayehu D, Abdul Qayum A, Haque TT, Taruselli MT, Paez PA, Pondicherry N, Barnstein BO, Hoeferlin LA, Chalfant CE and Ryan JJ: Lactic acid inhibits lipopolysaccharide-induced mast cell function by limiting glycolysis and ATP availability. J Immunol. 203:453–464. 2019. View Article : Google Scholar : PubMed/NCBI
20	Ideno M, Kobayashi M, Sasaki S, Futagi Y, Narumi K, Furugen A and Iseki K: Involvement of monocarboxylate transporter 1 (SLC16A1) in the uptake of l-lactate in human astrocytes. Life Sci. 192:110–114. 2018. View Article : Google Scholar : PubMed/NCBI
21	Curry JM, Tuluc M, Whitaker-Menezes D, Ames JA, Anantharaman A, Butera A, Leiby B, Cognetti DM, Sotgia F, Lisanti MP, et al: Cancer metabolism, stemness and tumor recurrence: MCT1 and MCT4 are functional biomarkers of metabolic symbiosis in head and neck cancer. Cell Cycle. 12:1371–1384. 2013. View Article : Google Scholar : PubMed/NCBI
22	Yuan Y, Guo-Qing P, Yan T, Hong-Lin Y, Gong-Hua H and Cai-Gao Z: A study of PKM2, PFK-1, and ANT1 expressions in cervical biopsy tissues in China. Med Oncol. 29:2904–2910. 2012. View Article : Google Scholar : PubMed/NCBI
23	Gooptu M, Whitaker-Menezes D, Sprandio J, Domingo-Vidal M, Lin Z, Uppal G, Gong J, Fratamico R, Leiby B, Dulau-Florea A, et al: Mitochondrial and glycolytic metabolic compartmentalization in diffuse large B-cell lymphoma. Semin Oncol. 44:204–217. 2017. View Article : Google Scholar : PubMed/NCBI
24	Sotgia F, Whitaker-Menezes D, Martinez-Outschoorn UE, Flomenberg N, Birbe RC, Witkiewicz AK, Howell A, Philp NJ, Pestell RG and Lisanti MP: Mitochondrial metabolism in cancer metastasis: Visualizing tumor cell mitochondria and the ‘reverse Warburg effect’ in positive lymph node tissue. Cell Cycle. 11:1445–1454. 2012. View Article : Google Scholar : PubMed/NCBI
25	Zhao Z, Han F, He Y, Yang S, Hua L, Wu J and Zhan W: Stromal-epithelial metabolic coupling in gastric cancer: Stromal MCT4 and mitochondrial TOMM20 as poor prognostic factors. Eur J Surg Oncol. 40:1361–1368. 2014. View Article : Google Scholar : PubMed/NCBI
26	Park SH, Lee AR, Choi K, Joung S, Yoon JB and Kim S: TOMM20 as a potential therapeutic target of colorectal cancer. BMB Rep. 52:712–717. 2019. View Article : Google Scholar : PubMed/NCBI
27	Duan K, Liu ZJ, Hu SQ, Huo HY, Xu ZR, Ruan JF, Sun Y, Dai LP, Yan CB, Xiong W, et al: Lactic acid induces lactate transport and glycolysis/OXPHOS interconversion in glioblastoma. Biochem Biophys Res Commun. 503:888–894. 2018. View Article : Google Scholar : PubMed/NCBI
28	Brown TP and Ganapathy V: Lactate/GPR81 signaling and proton motive force in cancer: Role in angiogenesis, immune escape, nutrition, and Warburg phenomenon. Pharmacol Ther. 206:1074512020. View Article : Google Scholar : PubMed/NCBI
29	Lee YJ, Shin KJ, Park SA, Park KS, Park S, Heo K, Seo YK, Noh DY, Ryu SH and Suh PG: G-protein-coupled receptor 81 promotes a malignant phenotype in breast cancer through angiogenic factor secretion. Oncotarget. 7:70898–70911. 2016. View Article : Google Scholar : PubMed/NCBI
30	Rancoule C, Guy JB, Vallard A, Ben Mrad M, Rehailia A and Magné N: 50th anniversary of cisplatin. Bull Cancer. 104:167–176. 2017. View Article : Google Scholar : PubMed/NCBI
31	Zhang K, Zhang B, Bai Y and Dai L: E2F1 promotes cancer cell sensitivity to cisplatin by regulating the cellular DNA damage response through miR-26b in esophageal squamous cell carcinoma. J Cancer. 11:301–310. 2020. View Article : Google Scholar : PubMed/NCBI
32	Shi S, Tan P, Yan B, Gao R, Zhao J, Wang J, Guo J, Li N and Ma Z: ER stress and autophagy are involved in the apoptosis induced by cisplatin in human lung cancer cells. Oncol Rep. 35:2606–2614. 2016. View Article : Google Scholar : PubMed/NCBI
33	Zheng AW, Chen YQ, Fang J, Zhang YL and Jia DD: Xiaoaiping combined with cisplatin can inhibit proliferation and invasion and induce cell cycle arrest and apoptosis in human ovarian cancer cell lines. Biomed Pharmacother. 89:1172–1177. 2017. View Article : Google Scholar : PubMed/NCBI
34	Xu O, Jia QJ, Wang JX and Shan C: A preliminary study on the function of GPR81 and TOMM20 in hypopharyngeal carcinoma. Chin J Otorhinolaryngol Skull Base Surg. 25:498–503. 2019.
35	Ashton TM, McKenna WG, Kunz-Schughart LA and Higgins GS: Oxidative phosphorylation as an emerging target in cancer therapy. Clin Cancer Res. 24:2482–2490. 2018. View Article : Google Scholar : PubMed/NCBI
36	Bottoni P and Scatena R: Mitochondrial metabolism in cancer. A tangled topic. Which role for proteomics? Adv Exp Med Biol. 1158:1–16. 2019. View Article : Google Scholar : PubMed/NCBI
37	Mor I, Cheung EC and Vousden KH: Control of glycolysis through regulation of PFK-1: Old friends and recent additions. Cold Spring Harb Symp Quant Biol. 76:211–216. 2011. View Article : Google Scholar : PubMed/NCBI
38	Marín-Hernández A, Gallardo-Pérez JC, López-Ramírez SY, García-García JD, Rodríguez-Zavala JS, Ruiz-Ramírez L, Gracia-Mora I, Zentella-Dehesa A, Sosa-Garrocho M, Macías-Silva M, et al: Casiopeina II-gly and bromo-pyruvate inhibition of tumor hexokinase, glycolysis, and oxidative phosphorylation. Arch Toxicol. 86:753–766. 2012. View Article : Google Scholar : PubMed/NCBI
39	Fan Z, Yu H, Cui N, Kong X, Liu X, Chang Y, Wu Y, Sun L and Wang G: ABT737 enhances cholangiocarcinoma sensitivity to cisplatin through regulation of mitochondrial dynamics. Exp Cell Res. 335:68–81. 2015. View Article : Google Scholar : PubMed/NCBI
40	Lottes RG, Newton DA, Spyropoulos DD and Baatz JE: Lactate as substrate for mitochondrial respiration in alveolar epithelial type II cells. Am J Physiol Lung Cell Mol Physiol. 308:L953–L961. 2015. View Article : Google Scholar : PubMed/NCBI
41	Yan C, Yang F, Zhou C, Chen X, Han X, Liu X, Ma H and Zheng W: MCT1 promotes the cisplatin-resistance by antagonizing Fas in epithelial ovarian cancer. Int J Clin Exp Pathol. 8:2710–2718. 2015.PubMed/NCBI
42	Hu Y and Zeng F: Expressions of GPR81, MCT1 and MCT4 in squamous carcinoma and their clinical significance. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 43:950–956. 2018.(In Chinese). PubMed/NCBI