miR‑199a‑3p/Sp1/LDHA axis controls aerobic glycolysis in testicular tumor cells

Zhou,Shihua; Min,Ziqian; Sun,Kang; Qu,Sijie; Zhou,Jinrun; Duan,Hongyan; Liu,Huawei; Liu,Xiaowen; Gong,Zhijun; Li,Dan

doi:10.3892/ijmm.2018.3771

miR‑199a‑3p/Sp1/LDHA axis controls aerobic glycolysis in testicular tumor cells

Authors:
- Shihua Zhou
- Ziqian Min
- Kang Sun
- Sijie Qu
- Jinrun Zhou
- Hongyan Duan
- Huawei Liu
- Xiaowen Liu
- Zhijun Gong
- Dan Li
View Affiliations

Affiliations: Department of Life Science, College of Biology, Hunan University, Changsha, Hunan 410082, P.R. China, Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R. China
Published online on: July 12, 2018 https://doi.org/10.3892/ijmm.2018.3771
Pages: 2163-2174

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

Aerobic glycolysis is one of the characteristics of tumor metabolism and contributes to the development of tumors. Studies have identified that microRNA (miRNA/miR) serves an important role in glucose metabolism of tumors. miR‑199a‑3p is a member of the miR‑199a family that controls the outcomes of cell survival and death processes, and previous studies have indicated that the expression of miR‑199a‑3p is low and may be an inhibitor in several cancer types, including testicular tumors. The present study discussed the role and underlying mechanism of miR‑199a‑3p in aerobic glycolysis of Ntera‑2 cells and identified its downstream factors. Firstly, miR‑199a‑3p exhibited an inhibitory effect on lactic acid production, glucose intake, and reactive oxygen species (ROS) and adenosine 5'‑triphosphate (ATP) levels in Ntera‑2 cells. Then, using bioinformatics, recombinant construction and a dual luciferase reporter gene system, transcription factor Specificity protein 1 (Sp1) was determined as the direct target of miR‑199a‑3p. Also, downregulation of Sp1 by RNA interference decreased lactic acid production, glucose intake, and ROS and ATP levels in Ntera‑2 cells. Subsequently, through a functional rescue experiment, it was identified that the overexpression of Sp1 may abate the inhibition of miR‑199a‑3p on glucose metabolism, with the exception of ATP level, suggesting a reciprocal association between Sp1 and miR‑199a‑3p. Finally, it was determined that miR‑199a‑3p overexpression and Sp1 knockdown decreased lactate dehydrogenase A (LDHA) protein expression, which indicated that LDHA is a downstream target of the miR‑199a‑3p/Sp1 signaling pathway. To additionally verify the regulation of LDHA expression by 199a‑3p/Sp1, a LDHA promoter reporter plasmid was generated and the high activity of the promoter, which contained 3 potential Sp1 binding elements, was confirmed. In addition, the overexpression of Sp1 led to the increased activity of the LDHA promoter, whereas knockdown of Sp1 exhibited the opposite effect. Therefore, the results of the present study demonstrated that miR‑199a‑3p can inhibit LDHA expression by downregulating Sp1, and provided mechanistic evidence supporting the existence of a novel miR‑199a‑3p/Sp1/LDHA axis and its critical contribution to aerobic glycolysis in testicular cancer cells

View Figures

View References

1	Pavlova NN and Thompson CB: The emerging hallmarks of cancer metabolism. Cell Metab. 23:27–47. 2016. View Article : Google Scholar : PubMed/NCBI
2	Lu J, Tan M and Cai Q: The Warburg effect in tumor progression: Mitochondrial oxidative metabolism as an anti-metastasis mechanism. Cancer Lett. 356:156–164. 2015. View Article : Google Scholar
3	Han T, Kang D, Ji D, Wang X, Zhan W, Fu M, Xin HB and Wang JB: How does cancer cell metabolism affect tumor migration and invasion? Cell Adh Migr. 7:395–403. 2013. View Article : Google Scholar : PubMed/NCBI
4	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
5	Lin HM, Nikolic I, Yang J, Castillo L, Deng N, Chan CL, Yeung NK, Dodson E, Elsworth B, Spielman C, et al: MicroRNAs as potential therapeutics to enhance chemosensitivity in advanced prostate cancer. Sci Rep. 8:78202018. View Article : Google Scholar : PubMed/NCBI
6	Sakurai K, Furukawa C, Haraguchi T, Inada K, Shiogama K, Tagawa T, Fujita S, Ueno Y, Ogata A, Ito M, et al: Micrornas miR-199a-5p and -3p target the brm subunit of swi/snf to generate a double-negative feedback loop in a variety of human cancers. Cancer Res. 71:1680–1689. 2011. View Article : Google Scholar
7	Gu S and Chan WY: Flexible and versatile as a chameleon-sophisticated functions of microRNA-199a. Int J Mol Sci. 13:8449–8466. 2012. View Article : Google Scholar : PubMed/NCBI
8	Hou J, Lin L, Zhou W, Wang Z, Ding G, Dong Q, Qin L, Wu X, Zheng Y, Yang Y, et al: Identification of miRNomes in human liver and hepatocellular carcinoma reveals miR-199a/b-3p as therapeutic target for hepatocellular carcinoma. Cancer Cell. 19:232–243. 2011. View Article : Google Scholar : PubMed/NCBI
9	Tsukigi M, Bilim V, Yuuki K, Ugolkov A, Naito S, Nagaoka A, Kato T, Motoyama T and Tomita Y: Re-expression of miR-199a suppresses renal cancer cell proliferation and survival by targeting GSK-3b. Cancer Lett. 315:189–197. 2012. View Article : Google Scholar
10	Minna E, Romeo P, De Cecco L, Dugo M, Cassinelli G, Pilotti S, Degl’Innocenti D, Lanzi C, Casalini P, Pierotti MA, et al: miR-199a-3p displays tumor suppressor functions in papillary thyroid carcinoma. Oncotarget. 5:2513–2528. 2014. View Article : Google Scholar : PubMed/NCBI
11	Qu F, Zheng J, Gan W, Lian H, He H, Li W, Yuan T, Yang Y, Li X, Ji C, et al: MiR-199a-3p suppresses proliferation and invasion of prostate cancer cells by targeting Smad1. Oncotarget. 8:52465–52473. 2017. View Article : Google Scholar : PubMed/NCBI
12	Zeng B, Shi W and Tan G: MiR-199a/b-3p inhibits gastric cancer cell proliferation via down-regulating PAK4/MEK/ERK signaling pathway. BMC Cancer. 18:342018. View Article : Google Scholar : PubMed/NCBI
13	Liu X, Duan H, Zhou S, Liu Z, Wu D, Zhao T, Xu S, Yang L and Li D: microRNA-199a-3p functions as tumor suppressor by regulating glucose metabolism in testicular germ cell tumors. Mol Med Rep. 14:2311–2320. 2016. View Article : Google Scholar : PubMed/NCBI
14	Chen BF, Gu S, Suen YK, Li L and Chan WY: microRNA-199a-3p, DNMT3A, and aberrant DNA methylation in testicular cancer. Epigenetics. 9:119–128. 2014. View Article : Google Scholar :
15	Deng Y, Zhao F, Hui L, Li X, Zhang D, Lin W, Chen Z and Ning Y: Suppressing miR-199a-3p by promoter methylation contributes to tumor aggressiveness and cisplatin resistance of ovarian cancer through promoting DDR1 expression. J Ovarian Res. 10:502017. View Article : Google Scholar : PubMed/NCBI
16	Li Q, Xia X, Ji J, Ma J, Tao L, Mo L and Chen W: MiR-199a-3p enhances cisplatin sensitivity of cholangiocarcinoma cells by inhibiting mTOR signaling pathway and expression of MDR1. Oncotarget. 8:33621–33630. 2017.PubMed/NCBI
17	Fan X, Zhou S, Zheng M, Deng X, Yi Y and Huang T: MiR-199a-3p enhances breast cancer cell sensitivity to cisplatin by downregulating TFAM (TFAM). Biomed Pharmacother. 88:507–514. 2017. View Article : Google Scholar : PubMed/NCBI
18	Agarwal V, Bell GW, Nam JW and Bartel DP: Predicting effective microRNA target sites in mammalian mRNAs. Elife. 4:2015. View Article : Google Scholar
19	Gori S, Porrozzi S, Roila F, Gatta G, De Giorgi U and Marangolo M: Germ cell tumours of the testis. Crit Rev Oncol Hematol. 53:141–164. 2005. View Article : Google Scholar : PubMed/NCBI
20	Winter C and Albers P: Testicular germ cell tumors: Pathogenesis, diagnosis and treatment. Nat Rev Endocrinol. 7:43–53. 2011. View Article : Google Scholar
21	von Eyben FE: Chromosomes, genes, and development of testicular germ cell tumors. Cancer Genet Cytogenet. 151:93–138. 2004. View Article : Google Scholar : PubMed/NCBI
22	Bezan A, Gerger A and Pichler M: MicroRNAs in testicular cancer: Implications for pathogenesis, diagnosis, prognosis and therapy. Anticancer Res. 34:2709–2713. 2014.PubMed/NCBI
23	Chieffi P and Chieffi S: Molecular biomarkers as potential targets for therapeutic strategies in human testicular germ cell tumors: An overview. J Cell Physiol. 228:1641–1646. 2013. View Article : Google Scholar : PubMed/NCBI
24	Viatori M: Testicular cancer. Semio Oncol Nurs. 28:180–189. 2012. View Article : Google Scholar
25	Ferreira LM, Hebrant A and Dumont JE: Metabolic reprogramming of the tumor. Oncogene. 31:3999–4011. 2012. View Article : Google Scholar : PubMed/NCBI
26	Porporato PE, Dhup S, Dadhich RK, Copetti T and Sonveaux P: Anticancer targets in the glycolytic metabolism of tumors: A comprehensive review. Front Pharmacol. 2:492011. View Article : Google Scholar : PubMed/NCBI
27	Xiao X, Huang X, Ye F, Chen B, Song C, Wen J, Zhang Z, Zheng G, Tang H and Xie X: The miR-34a-LDHA axis regulates glucose metabolism and tumor growth in breast cancer. Sci Rep. 6:217352016. View Article : Google Scholar : PubMed/NCBI
28	Rizwan A, Serganova I, Khanin R, Karabeber H, Ni X, Thakur S, Zakian KL, Blasberg R and Koutcher JA: Relationships between LDH-A, lactate, and metastases in 4T1 breast tumors. Clin Cancer Res. 19:5158–5169. 2013. View Article : Google Scholar : PubMed/NCBI
29	Qiu H, Jackson AL, Kilgore JE, Zhong Y, Chan LL, Gehrig PA, Zhou C and Bae-Jump VL: JQ1 suppresses tumor growth through downregulating LDHA in ovarian cancer. Oncotarget. 6:6915–6930. 2015. View Article : Google Scholar : PubMed/NCBI
30	Wang J, Wang H, Liu A, Fang C, Hao J and Wang Z: Lactate dehydrogenase A negatively regulated by miRNAs promotes aerobic glycolysis and is increased in colorectal cancer. Oncotarget. 6:19456–19468. 2015.PubMed/NCBI
31	Otero-Albiol D and Felipe-Abrio B: MicroRNA regulating metabolic reprogramming in tumor cells: New tumor markers. Cancer Transl Med. 2:175–81. 2016. View Article : Google Scholar
32	Zou S, Gu Z, Ni P, Liu X, Wang J and Fan Q: SP1 plays a pivotal role for basal activity of TIGAR promoter in liver cancer cell lines. Mol Cell Biochem. 359:17–23. 2012. View Article : Google Scholar
33	Archer MC: Role of sp transcription factors in the regulation of cancer cell metabolism. Genes Cancer. 2:712–719. 2011. View Article : Google Scholar : PubMed/NCBI
34	Hedrick E, Cheng Y, Jin UH, Kim K and Safe S: Specificity protein (Sp) transcription factors Sp1, Sp3 and Sp4 are non-oncogene addiction genes in cancer cells. Oncotarget. 7:22245–22256. 2016. View Article : Google Scholar : PubMed/NCBI
35	Beishline K and Azizkhan-Clifford J: Sp1 and the ‘hallmarks of cancer’. FEBS J. 282:224–258. 2015. View Article : Google Scholar
36	Vizcaíno C, Mansilla S and Portugal J: Sp1 transcription factor: A long-standing target in cancer chemotherapy. Pharmacol Ther. 152:111–124. 2015. View Article : Google Scholar : PubMed/NCBI