Effect of multidrug resistance 1/P-glycoprotein on the hypoxia-induced multidrug resistance of human laryngeal cancer cells

Li,Dawei; Zhou,Liang; Huang,Jiameng; Xiao,Xiyan

doi:10.3892/ol.2016.4749

Effect of multidrug resistance 1/P-glycoprotein on the hypoxia-induced multidrug resistance of human laryngeal cancer cells

Authors:
- Dawei Li
- Liang Zhou
- Jiameng Huang
- Xiyan Xiao
View Affiliations

Affiliations: Department of Otolaryngology ‑ Head and Neck Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai 200233, P.R. China, Department of Otolaryngology ‑ Head and Neck Surgery, Affiliated Eye and Ear, Nose and Throat Hospital of Fudan University, Shanghai 200031, P.R. China
Published online on: June 17, 2016 https://doi.org/10.3892/ol.2016.4749
Pages: 1569-1574

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

In a previous study, it was demonstrated that hypoxia upregulated the multidrug resistance (MDR) of laryngeal cancer cells to chemotherapeutic drugs, with multidrug resistance 1 (MDR1)/P-glycoprotein (P‑gp) expression also being upregulated. The present study aimed to investigate the role and mechanism of MDR1/P‑gp on hypoxia-induced MDR in human laryngeal carcinoma cells. The sensitivity of laryngeal cancer cells to multiple drugs and cisplatin‑induced apoptosis was determined by CCK‑8 assay and Annexin‑V/propidium iodide staining analysis, respectively. The accumulation of rhodamine 123 (Rh123) in the cells served as an estimate of drug accumulation and was evaluated by flow cytometry (FCM). MDR1/P-gp expression was inhibited using interference RNA, and the expression of the MDR1 gene was analyzed using reverse transcription-quantitative polymerase chain reaction and western blotting. As a result, the sensitivity to multiple chemotherapeutic agents and the apoptosis rate of the hypoxic laryngeal carcinoma cells increased following a decrease in MDR1/P‑gp expression (P<0.05). Additionally, FCM analysis of fluorescence intensity indicated that the downregulated expression of MDR1/P‑gp markedly increased intracellular Rh123 accumulation (P<0.05). Such results suggest that MDR1/P‑gp serves an important role in regulating hypoxia-induced MDR in human laryngeal carcinoma cells through a decrease in intracellular drug accumulation.

View Figures

View References

1	Chu EA and Kim YJ: Laryngeal cancer: Diagnosis and preoperative work-up. Otolaryngol Clin North Am. 41:673–695, v. 2008. View Article : Google Scholar : PubMed/NCBI
2	Liu HC, Chen GG, Vlantis AC, Tong MC and van Hasselt CA: Chemotherapy for laryngeal cancer - an apoptotic approach. Curr Drug Targets. 9:878–886. 2008. View Article : Google Scholar : PubMed/NCBI
3	Bar-Ad V, Palmer J, Yang H, Cognetti D, Curry J, Luginbuhl A, Tuluc M, Campling B and Axelrod R: Current management of locally advanced head and neck cancer: The combination of chemotherapy with locoregional treatments. Semin Oncol. 41:798–806. 2014. View Article : Google Scholar : PubMed/NCBI
4	Wang GL, Jiang BH, Rue EA and Semenza GL: Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O₂ tension. Proc Natl Acad Sci USA. 92:5510–5514. 1995. View Article : Google Scholar : PubMed/NCBI
5	Wohlkoenig C, Leithner K, Deutsch A, Hrzenjak A, Olschewski A and Olschewski H: Hypoxia-induced cisplatin resistance is reversible and growth rate independent in lung cancer cells. Cancer Lett. 308:134–143. 2011. View Article : Google Scholar : PubMed/NCBI
6	Yoshiba S, Ito D, Nagumo T, Shirota T, Hatori M and Shintani S: Hypoxia induces resistance to 5-fluorouracil in oral cancer cells via G(1) phase cell cycle arrest. Oral Oncol. 45:109–115. 2009. View Article : Google Scholar : PubMed/NCBI
7	Huang L, Ao Q, Zhang Q, Yang X, Xing H, Li F, Chen G, Zhou J, Wang S, Xu G, et al: Hypoxia induced paclitaxel resistance in human ovarian cancers via hypoxia-inducible factor 1alpha. J Cancer Res Clin Oncol. 136:447–456. 2010. View Article : Google Scholar : PubMed/NCBI
8	Li DW, Dong P, Wang F, Chen XW, Xu CZ and Zhou L: Hypoxia induced multidrug resistance of laryngeal cancer cells via hypoxia-inducible factor-1α. Asian Pac J Cancer Prev. 14:4853–4858. 2013. View Article : Google Scholar : PubMed/NCBI
9	Li DW, Gao S, Shen B and Dong P: Effect of apoptotic and proliferative indices, P-glycoprotein and survivin expression on prognosis in laryngeal squamous cell carcinoma. Med Oncol. 28(Suppl 1): S333–S340. 2011. View Article : Google Scholar : PubMed/NCBI
10	Li L, Jiang AC, Dong P, Wan Y and Yu ZW: The characteristics of Hep-2 cell with multiple drug resistance induced by Taxol. Otolaryngol Head Neck Surg. 137:659–664. 2007. View Article : Google Scholar : PubMed/NCBI
11	Zhigang H, Qi Z, Jugao F, Xiaohong C, Wei Z, Hong W, Hu H, Na M, Zheng Y and Demin H: Reverse multidrug resistance in laryngeal cancer cells by knockdown MDR1 gene expression. J Otolaryngol Head Neck Surg. 38:440–448. 2009.PubMed/NCBI
12	Xie J, Li DW, Chen XW, Wang F and Dong P: Expression and significance of hypoxia-inducible factor-1α and MDR1/P-glycoprotein in laryngeal carcinoma tissue and hypoxic Hep-2 cells. Oncol Lett. 6:232–238. 2013.PubMed/NCBI
13	Sowter HM, Raval RR, Moore JW, Ratcliffe PJ and Harris AL: Predominant role of hypoxia-inducible transcription factor (Hif)-1alpha versus Hif-2alpha in regulation of the transcriptional response to hypoxia. Cancer Res. 63:6130–6134. 2003.PubMed/NCBI
14	Li DW, Zhou L, Jin B, Xie J and Dong P: Expression and significance of hypoxia-inducible factor-1α and survivin in laryngeal carcinoma tissue and cells. Otolaryngol Head Neck Surg. 148:75–81. 2013. View Article : Google Scholar : PubMed/NCBI
15	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
16	Ludescher C, Thaler J, Drach D, Drach J, Spitaler M, Gattringer C, Huber H and Hofmann J: Detection of activity of P-glycoprotein in human tumour samples using rhodamine 123. Br J Haematol. 82:161–168. 1992. View Article : Google Scholar : PubMed/NCBI
17	Zhu H, Chen XP, Luo SF, Guan J, Zhang WG and Zhang BX: Involvement of hypoxia-inducible factor-1-alpha in multidrug resistance induced by hypoxia in HepG2 cells. J Exp Clin Cancer Res. 24:565–574. 2005.PubMed/NCBI
18	Liu L, Ning X, Sun L, Zhang H, Shi Y, Guo C, Han S, Liu J, Sun S, Han Z, et al: Hypoxia-inducible factor-1 alpha contributes to hypoxia-induced chemoresistance in gastric cancer. Cancer Sci. 99:121–128. 2008.PubMed/NCBI
19	Huang C, Xu D, Xia Q, Wang P, Rong C and Su Y: Reversal of P-glycoprotein-mediated multidrug resistance of human hepatic cancer cells by Astragaloside II. J Pharm Pharmacol. 64:1741–1750. 2012. View Article : Google Scholar : PubMed/NCBI
20	Hao YX, He ZW, Zhu JH, Shen Q, Sun JZ, Du N and Xiao WH: Reversal of multidrug resistance in renal cell carcinoma by short hairpin RNA targeting MDR1 gene. Chin Med J (Engl). 125:2741–2745. 2012.PubMed/NCBI
21	Jia M, Hu J, Li W, Su P, Zhang H, Zhang X and Zhou G: Trps1 is associated with the multidrug resistance of osteosarcoma by regulating MDR1 gene expression. FEBS Lett. 588:801–810. 2014. View Article : Google Scholar : PubMed/NCBI
22	Wartenberg M, Ling FC, Müschen M, Klein F, Acker H, Gassmann M, Petrat K, Pütz V, Hescheler J and Sauer H: Regulation of the multidrug resistance transporter P-glycoprotein in multicellular tumor spheroids by hypoxia-inducible factor (HIF-1) and reactive oxygen species. FASEB J. 17:503–505. 2003.PubMed/NCBI
23	Xia S, Yu SY, Yuan XL and Xu SP: Effects of hypoxia on expression of P-glycoprotein and multidrug resistance protein in human lung adenocarcinoma A549 cell line. Zhonghua Yi Xue Za Zhi. 84:663–666. 2004.(In Chinese). PubMed/NCBI
24	Song X, Liu X, Chi W, Liu Y, Wei L, Wang X and Yu J: Hypoxia-induced resistance to cisplatin and doxorubicin in non-small cell lung cancer is inhibited by silencing of HIF-1alpha gene. Cancer Chemother Pharmacol. 58:776–784. 2006. View Article : Google Scholar : PubMed/NCBI
25	Kimura Y, Matsuo M, Takahashi K, Saeki T, Kioka N, Amachi T and Ueda K: ATP hydrolysis-dependent multidrug efflux transporter: MDR1/P-glycoprotein. Curr Drug Metab. 5:1–10. 2004. View Article : Google Scholar : PubMed/NCBI
26	Ueda K: Molecular mechanisms for drug transport by MDR1/P-glycoprotein. Tanpakushitsu Kakusan Koso. 42:1263–1272. 1997.(In Japanese). PubMed/NCBI