Glucose transporter-1 expression in CD133+ laryngeal carcinoma Hep-2 cells

Chen,Xiao-Hong; Bao,Yang-Yang; Zhou,Shui-Hong; Wang,Qin-Ying; Wei,Yan; Fan,Jun

doi:10.3892/mmr.2013.1740

Glucose transporter-1 expression in CD133+ laryngeal carcinoma Hep-2 cells

Authors:
- Xiao-Hong Chen
- Yang-Yang Bao
- Shui-Hong Zhou
- Qin-Ying Wang
- Yan Wei
- Jun Fan
View Affiliations

Affiliations: Department of Otolaryngology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China
Published online on: October 18, 2013 https://doi.org/10.3892/mmr.2013.1740
Pages: 1695-1700

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

CD133 is a useful putative marker of cancer stem cells (CSCs) in human laryngeal tumors. Numerous studies have demonstrated that CD133+ CSCs possess higher clonogenicity, invasiveness and tumorigenesis compared with CD133- cells. Recently, interest in the Warburg effect in the microenvironment of CSCs has escalated. The Warburg effect dictates that cancer cells rely on glycolysis rather than oxidative phosphorylation under aerobic conditions. In numerous cancer cells, glucose is used mainly for the glycolytic pathway. Stem cells express high levels of glycolytic enzymes and rely mostly on glycolysis to meet their energy demands. Glucose is transported through cell membranes by glucose transporters (Glut). Studies of Glut-1 expression in CSCs are limited. In the present study, we investigated the proliferation of CD133+ Hep-2 cells and whether Glut-1 is expressed in laryngeal carcinoma CD133+ Hep-2 cells. Real-time reverse transcription-polymerase chain reaction (RT-PCR) demonstrated that the size of the CD133 product was 213 bp. Dissociation curve analysis demonstrated only the expected peaks at 82.1˚C for CD133. The mean ΔCt of CD133 expression was 10.98. Prior to isolation, the CD133+ fraction was 1.2% by fluorescence-activated cell sorting (FACS) analysis. Following isolation, the CD133+ fraction was increased to 76.1%. Successive tests also demonstrated that cells grew well following isolation. The proliferation of CD133+ and CD133- cells was not different during the first 3 days (P>0.05). From day 4, the proliferation capacity of CD133+ cells in vitro was higher than that of CD133- cells (P<0.05). The mean ΔCt of Glut-1 mRNA expression was 1.78 for CD133+ cells and 1.00 for CD133- cells (P<0.05). The mean Glut-1 protein values in CD133+ and CD133- Hep-2 cells relative to β-tubulin were 0.48 ± 0.02 and 0.21 ± 0.03 (µg/µl), respectively (P<0.05). In conclusion, CD133+ cells demonstrated higher proliferation. Glut-1 mRNA and protein levels were higher in CD133+ than in CD133- cells. Our results suggest that Glut-1 is important in the energy supply of laryngeal CD133+ Hep-2 cells and Glut-1 may represent a potential therapeutic target for the inhibition of the proliferation of laryngeal CSCs.

View Figures

View References

1	Yang JP, Liu Y, Zhong W, Yu D, Wen LJ and Jin CS: Chemoresistance of CD133⁺ cancer stem cells in laryngeal carcinoma. Chin Med J (Engl). 124:1055–1060. 2011.PubMed/NCBI
2	Chen H, Zhou L, Dou T, Wan G, Tang H and Tian J: BMI1’S maintenance of the proliferative capacity of laryngeal cancer stem cells. Head Neck. 33:1115–1125. 2011.
3	Wei XD, Zhou L, Cheng L, Tian J, Jiang JJ and Maccallum J: In vivo investigation of CD133 as a putative marker of cancer stem cells in Hep-2 cell line. Head Neck. 31:94–101. 2009. View Article : Google Scholar : PubMed/NCBI
4	Zhou L, Wei X, Cheng L, Tian J and Jiang JJ: CD133, one of the markers of cancer stem cells in Hep-2 cell line. Laryngoscope. 117:455–460. 2007. View Article : Google Scholar : PubMed/NCBI
5	Piao LS, Hur W, Kim TK, et al: CD133⁺ liver cancer stem cells modulate radioresistance in human hepatocellular carcinoma. Cancer Lett. 315:129–137. 2012.
6	Ke CC, Liu RS, Yang AH, et al: CD133-expressing thyroid cancer cells are undifferentiated, radioresistant and survive radioiodide therapy. Eur J Nucl Med Mol Imaging. 40:61–71. 2013. View Article : Google Scholar : PubMed/NCBI
7	Bao S, Wu Q, McLendon RE, et al: Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 444:756–760. 2006. View Article : Google Scholar : PubMed/NCBI
8	Liu YP, Yang CJ, Huang MS, et al: Cisplatin selects for multidrug-resistant CD133⁺ cells in lung adenocarcinoma by activating Notch signaling. Cancer Res. 73:406–416. 2013. View Article : Google Scholar : PubMed/NCBI
9	Neth P, Ries C, Karow M, Egea V, Ilmer M and Jochum M: The Wnt signal transduction pathway in stem cells and cancer cells: influence on cellular invasion. Stem Cell Rev. 3:18–29. 2007. View Article : Google Scholar : PubMed/NCBI
10	Huang D, Gao Q, Guo L, et al: Isolation and identification of cancer stem-like cells in esophageal carcinoma cell lines. Stem Cells Dev. 18:465–473. 2009. View Article : Google Scholar : PubMed/NCBI
11	Gangopadhyay S, Nandy A, Hor P and Mukhopadhyay A: Breast cancer stem cells: a novel therapeutic target. Clin Breast Cancer. 13:7–15. 2013. View Article : Google Scholar
12	Ping YF, Yao XH, Jiang JY, et al: The chemokine CXCL12 and its receptor CXCR4 promote glioma stem cell-mediated VEGF production and tumour angiogenesis via PI3K/AKT signalling. J Pathol. 224:344–354. 2011. View Article : Google Scholar : PubMed/NCBI
13	Bensinger SJ and Christofk HR: New aspects of the Warburg effect in cancer cell biology. Semin Cell Dev Biol. 23:352–361. 2012. View Article : Google Scholar : PubMed/NCBI
14	Zhao K, Luo XM, Zhou SH, et al: ¹⁸F-fluorodeoxyglucose positron emission tomography/computed tomography as an effective diagnostic workup in cervical metastasis of carcinoma from an unknown primary tumor. Cancer Biother Radiopharm. 27:685–693. 2012. View Article : Google Scholar
15	Hammoudi N, Ahmed KB, Garcia-Prieto C and Huang P: Metabolic alterations in cancer cells and therapeutic implications. Chin J Cancer. 30:508–525. 2011. View Article : Google Scholar : PubMed/NCBI
16	Varum S, Rodrigues AS, Moura MB, et al: Energy metabolism in human pluripotent stem cells and their differentiated counterparts. PLoS One. 6:e209142011. View Article : Google Scholar : PubMed/NCBI
17	Han MW, Lee HJ, Cho KJ, et al: Role of FDG-PET as a biological marker for predicting the hypoxic status of tongue cancer. Head Neck. 34:1395–1402. 2012. View Article : Google Scholar : PubMed/NCBI
18	Li XF, Ma Y, Sun X, Humm JL, Ling CC and O’Donoghue JA: High 18F-FDG uptake in microscopic peritoneal tumors requires physiologic hypoxia. J Nucl Med. 51:632–638. 2010. View Article : Google Scholar : PubMed/NCBI
19	Alakus H, Batur M, Schmidt M, et al: Variable 18F-fluorodeoxyglucose uptake in gastric cancer is associated with different levels of GLUT-1 expression. Nucl Med Commun. 31:532–538. 2010.PubMed/NCBI
20	Nagamatsu A, Umesaki N, Li L and Tanaka T: Use of 18F-fluorodeoxyglucose positron emission tomography for diagnosis of uterine sarcomas. Oncol Rep. 23:1069–1076. 2010.PubMed/NCBI
21	Zhou S, Wang S, Wu Q, Fan J and Wang Q: Expression of glucose transporter-1 and -3 in the head and neck carcinoma--the correlation of the expression with the biological behaviors. ORL J Otorhinolaryngol Relat Spec. 70:189–194. 2008. View Article : Google Scholar : PubMed/NCBI
22	Li LF, Zhou SH, Zhao K, et al: Clinical significance of FDG single-photon emission computed tomography: Computed tomography in the diagnosis of head and neck cancers and study of its mechanism. Cancer Biother Radiopharm. 23:701–714. 2008. View Article : Google Scholar : PubMed/NCBI
23	Mai HM, Zheng JW, Wang YA, et al: CD133 selected stem cells from proliferating infantile hemangioma and establishment of an in vivo mice model of hemangioma. Chin Med J (Engl). 126:88–94. 2013.PubMed/NCBI
24	Loda M, Xu X, Pession A, Vortmeyer A and Giangaspero F: Membranous expression of glucose transporter-1 protein (GLUT-1) in embryonal neoplasms of the central nervous system. Neuropathol Appl Neurobiol. 26:91–97. 2000. View Article : Google Scholar : PubMed/NCBI
25	Wu XH, Chen SP, Mao JY, Ji XX, Yao HT and Zhou SH: Expression and significance of hypoxia-inducible factor-1α and glucose transporter-1 in laryngeal carcinoma. Oncol Lett. 5:261–266. 2013.
26	Xu YY, Bao YY, Zhou SH and Fan J: Effect on the expression of MMP-2, MT-MMP in laryngeal carcinoma Hep-2 cell line by antisense glucose transporter-1. Arch Med Res. 43:395–401. 2012. View Article : Google Scholar : PubMed/NCBI
27	Zhou SH, Fan J, Chen XM, Cheng KJ and Wang SQ: Inhibition of cell proliferation and glucose uptake in human laryngeal carcinoma cells by antisense oligonucleotides against glucose transporter-1. Head Neck. 31:1624–1633. 2009. View Article : Google Scholar : PubMed/NCBI
28	Korkeila E, Jaakkola PM, Syrjänen K, Pyrhönen S and Sundström J: Pronounced tumour regression after radiotherapy is associated with negative/weak glucose transporter-1 expression in rectal cancer. Anticancer Res. 31:311–315. 2011.PubMed/NCBI
29	Chiba I, Ogawa K, Morioka T, et al: Clinical significance of GLUT-1 expression in patients with esophageal cancer treated with concurrent chemoradiotherapy. Oncol Lett. 2:21–28. 2011.PubMed/NCBI
30	Kunkel M, Moergel M, Stockinger M, et al: Overexpression of GLUT-1 is associated with resistance to radiotherapy and adverse prognosis in squamous cell carcinoma of the oral cavity. Oral Oncol. 43:796–803. 2007. View Article : Google Scholar : PubMed/NCBI
31	Eckert AW, Kappler M, Schubert J and Taubert H: Correlation of expression of hypoxia-related proteins with prognosis in oral squamous cell carcinoma patients. Oral Maxillofac Surg. 16:189–196. 2012. View Article : Google Scholar : PubMed/NCBI
32	Rademakers SE, Lok J, van der Kogel AJ, Bussink J and Kaanders JH: Metabolic markers in relation to hypoxia; staining patterns and colocalization of pimonidazole, HIF-1α, CAIX, LDH-5, GLUT-1, MCT1 and MCT4. BMC Cancer. 11:1672011.PubMed/NCBI
33	Cao X, Fang L, Gibbs S, et al: Glucose uptake inhibitor sensitizes cancer cells to daunorubicin and overcomes drug resistance in hypoxia. Cancer Chemother Pharmacol. 59:495–505. 2007. View Article : Google Scholar : PubMed/NCBI