Differential expression of multidrug resistance‑related proteins in adriamycin‑resistant (pumc‑91/ADM) and parental (pumc‑91) human bladder cancer cell lines

Zhao,Man; Yu,Shuliang; Zhang,Man

doi:10.3892/mmr.2016.5806

Differential expression of multidrug resistance‑related proteins in adriamycin‑resistant (pumc‑91/ADM) and parental (pumc‑91) human bladder cancer cell lines

Authors:
- Man Zhao
- Shuliang Yu
- Man Zhang
View Affiliations

Affiliations: Clinical Laboratory Medicine, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, P.R. China
Published online on: October 5, 2016 https://doi.org/10.3892/mmr.2016.5806
Pages: 4741-4746

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

Multidrug resistance (MDR) is the major obstacle to bladder cancer chemotherapy. Several mechanisms have been implicated in the development of MDR, including extrusion of the drug by cell membrane pumps, associated with P‑glycoprotein (P‑gp) and multidrug resistance‑associated protein (MRP); increased DNA damage repair, associated with topoisomerase II (Topo II); suppression of drug‑induced apoptosis, associated with p53; and regulation of cancer cell growth, associated with vascular endothelial growth factor (VEGF). In the present study, the expression levels of these five markers were detected in an adriamycin (ADM)‑resistant human bladder cancer cell line (pumc‑91/ADM) and its parental cell line (pumc‑91), in order to determine which marker is more important, or whether all of them participate in drug resistance. The expression levels of P‑gp, MRP, Topo II, VEGF and p53 were measured in the two cell lines by reverse transcription‑quantitative polymerase chain reaction, western blotting and immunohistochemistry. A significant increase in P‑gp, MRP and VEGF, and a decrease in Topo II mRNA expression were detected in the pumc‑91/ADM drug‑resistant cell line compared with the pumc‑91 cell line; however, no difference in p53 mRNA expression was detected between the cells. In pumc‑91/ADM cells, the protein expression levels of P‑gp and MRP were upregulated, whereas Topo II was significantly decreased. However, no marked differences in p53 or VEGF expression were detected between the two cell lines at the protein level. The cytoplasmic and cell membrane localization of P‑gp and MRP, the cytoplasmic localization of VEGF, and the nuclear localization of p53 and Topo II were confirmed in the two cell lines. The present study detected increased P‑gp and MRP, and reduced Topo II expression in pumc‑91/ADM cells compared with pumc‑91 cells; however, no difference was detected in p53 and VEGF expression between the cell lines. In conclusion, a significant upregulation of MRP and downregulation of Topo II were detected in the ADM‑resistant human bladder cancer cell line (pumc‑91/ADM) compared with in the parental cell line (pumc‑91).

View Figures

View References

1	Lei T, Zhao X, Jin S, Meng Q, Zhou H and Zhang M: Discovery of potential bladder cancer biomarkers by comparative urine proteomics and analysis. Clin Genitourin Cancer. 11:56–62. 2013. View Article : Google Scholar : PubMed/NCBI
2	Meng Q, Lei T and Zhang M, Zhao J, Zhao XH and Zhang M: Identification of proteins differentially expressed in adriamycin-resistant (pumc-91/ADM) and parental (pumc-91) human bladder cancer cell lines by proteome analysis. J Cancer Res Clin Oncol. 139:509–519. 2013. View Article : Google Scholar : PubMed/NCBI
3	Sacerdote C, Guarrera S, Ricceri F, Pardini B, Polidoro S, Allione A, Critelli R, Russo A, Andrew AS, Ye Y, et al: Polymorphisms in the XRCC1 gene modify survival of bladder cancer patients treated with chemotherapy. Int J Cancer. 133:2004–2009. 2013. View Article : Google Scholar : PubMed/NCBI
4	Zeekpudsa P, Kukongviriyapan V, Senggunprai L, Sripa B and Prawan A: Suppression of NAD(P)H-quinone oxidoreductase 1 enhanced the susceptibility of cholangiocarcinoma cells to chemotherapeutic agents. J Exp Clin Cancer Res. 33:112014. View Article : Google Scholar : PubMed/NCBI
5	Zandvliet M, Teske E and Schrickx JA: Multi-drug resistance in a canine lymphoid cell line due to increased P-glycoprotein expression, a potential model for drug-resistant canine lymphoma. Toxicol In Vitro. 28:1498–1506. 2014. View Article : Google Scholar : PubMed/NCBI
6	Li W and Song M: Expression of multidrug resistance proteins in invasive ductal carcinoma of the breast. Oncol Lett. 8:2103–2109. 2014.PubMed/NCBI
7	Semiglazova TIu, Klimenko VV, Filatova LV, Chubenko VA, Krivorot'ko PV, Ivanov VG, Turkevich EA, Ivantsov AO, Novikov SN, Semiglazov VV, et al: Markers of effectiveness of preoperative taxane-based chemotherapy for locally advanced breast cancer. Vopr Onkol. 59:363–367. 2013.(In Russian). PubMed/NCBI
8	Grosset AA, Labrie M, Gagné D, Vladoiu MC, Gaboury L, Doucet N and St-Pierre Y: Cytosolic galectin-7 impairs p53 functions and induces chemoresistance in breast cancer cells. BMC Cancer. 14:8012014. View Article : Google Scholar : PubMed/NCBI
9	Ki CS, Lin TY, Korc M and Lin CC: Thiol-ene hydrogels as desmoplasia-mimetic matrices for modeling pancreatic cancer cell growth, invasion, and drug resistance. Biomaterials. 35:9668–9677. 2014. View Article : Google Scholar : PubMed/NCBI
10	Zhang M, Jin S and Zhang M: Establishing human multi-drug resistant bladder cancer cell lines Pumc-91/ADM and evaluating its biological characteristics. J Med Res. 38:70–72. 2009.
11	Yu S, Meng Q, Hu H and Zhang M: Correlation of ANXA1 expression with drug resistance and relapse in bladder cancer. Int J Clin Exp Pathol. 7:5538–5548. 2014.PubMed/NCBI
12	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
13	Hu H, Meng Q, Lei T and Zhang M: Nucleophosmin1 associated with drug resistance and recurrence of bladder cancer. Clin Exp Med. 15:361–369. 2015. View Article : Google Scholar : PubMed/NCBI
14	Pappas JJ, Petropoulos S, Suderman M, Iqbal M, Moisiadis V, Turecki G, Matthews SG and Szyf M: The multidrug resistance 1 gene Abcb1 in brain and placenta: Comparative analysis in human and guinea pig. PLoS One. 9:e1111352014. View Article : Google Scholar : PubMed/NCBI
15	Yu P, Du Y, Cheng X, Yu Q, Huang L and Dong R: Expression of multidrug resistance-associated proteins and their relation to postoperative individualized chemotherapy in gastric cancer. World J Surg Oncol. 12:3072014. View Article : Google Scholar : PubMed/NCBI
16	Kodiakov DS, Klimachëv VV, Avdalian AM, Bobrov IP, Lazarev AF, Lushnikova EL and Nepomiashchikh LM: Topoisomerase IIa expression in correlation with clinical and morphological parameters and proliferation (based on argyrophilic proteins of nucleolar organizer regions and Ki-67 antigen) in lung adenocarcinoma. Vopr Onkol. 60:63–68. 2014.(In Russian). PubMed/NCBI
17	Llovet JM: Focal gains of VEGFA: Candidate predictors of sorafenib response in hepatocellular carcinoma. Cancer Cell. 25:560–562. 2014. View Article : Google Scholar : PubMed/NCBI
18	Tonigold M, Rossmann A, Meinold M, Bette M, Märken M, Henkenius K, Bretz AC, Giel G, Cai C, Rodepeter FR, et al: A cisplatin-resistant head and neck cancer cell line with cytoplasmic p53(mut) exhibits ATP-binding cassette transporter upregulation and high glutathione levels. J Cancer Res Clin Oncol. 140:1689–1704. 2014. View Article : Google Scholar : PubMed/NCBI
19	An J, Wang X, Guo P, Zhong Y, Zhang X and Yu Z: Hexabromocyclododecane and polychlorinated biphenyls increase resistance of hepatocellular carcinoma cells to cisplatin through the phosphatidylinositol 3-kinase/protein kinase B pathway. Toxicol Lett. 229:265–272. 2014. View Article : Google Scholar : PubMed/NCBI
20	Kovalev AA, Tsvetaeva DA and Grudinskaja TV: Role of ABC-cassette transporters (MDR1, MRP1, BCRP) in the development of primary and acquired multiple drug resistance in patients with early and metastatic breast cancer. Exp Oncol. 35:287–290. 2013.PubMed/NCBI
21	Lee YK, Lin TH, Chang CF and Lo YL: Galectin-3 silencing inhibits epirubicin-induced ATP binding cassette transporters and activates the mitochondrial apoptosis pathway via β-catenin/GSK-3β modulation in colorectal carcinoma. PLoS One. 8:e824782013. View Article : Google Scholar : PubMed/NCBI
22	Copsel S, Garcia C, Diez F, Vermeulem M, Baldi A, Bianciotti LG, Russel FG, Shayo C and Davio C: Multidrug resistance protein 4 (MRP4/ABCC4) regulates cAMP cellular levels and controls human leukemia cell proliferation and differentiation. J Biol Chem. 286:6979–6988. 2011. View Article : Google Scholar : PubMed/NCBI
23	Su W and Pasternak GW: The role of multidrug resistance-associated protein in the blood-brain barrier and opioid analgesia. Synapse. 67:609–619. 2013. View Article : Google Scholar : PubMed/NCBI
24	Yu P, Du Y, Cheng X, Yu Q, Huang L and Dong R: Expression of multidrug resistance-associated proteins and their relation to postoperative individualized chemotherapy in gastric cancer. World J Surg Oncol. 12:3072014. View Article : Google Scholar : PubMed/NCBI
25	Tsang WP, Kong SK and Kwok TT: Epidermal growth factor induction of resistance to topoisomerase II toxins in human squamous carcinoma A431 cells. Oncol Rep. 16:789–793. 2006.PubMed/NCBI
26	Campiglio M, Somenzi G, Olgiati C, Beretta G, Balsari A, Zaffaroni N, Valagussa P and Ménard S: Role of proliferation in HER2 status predicted response to doxorubicin. Int J Cancer. 105:568–573. 2003. View Article : Google Scholar : PubMed/NCBI
27	Fillmore CM, Xu C, Desai PT, Berry JM, Rowbotham SP, Lin YJ, Zhang H, Marquez VE, Hammerman PS, Wong KK and Kim CF: EZH2 inhibition sensitizes BRG1 and EGFR mutant lung tumours to TopoII inhibitors. Nature. 520:239–242. 2015. View Article : Google Scholar : PubMed/NCBI
28	Chin KV and Kong AN: Application of DNA microarrays in pharmacogenomics and toxicogenomics. Pharm Res. 19:1773–1778. 2002. View Article : Google Scholar : PubMed/NCBI
29	Garufi A and D'Orazi G: High glucose dephosphorylates serine 46 and inhibits p53 apoptotic activity. J Exp Clin Cancer Res. 33:792014. View Article : Google Scholar : PubMed/NCBI
30	Lin J, Shen A, Chen H, Liao J, Xu T, Liu L, Lin J and Peng J: Nitidine chloride inhibits hepatic cancer growth via modulation of multiple signaling pathways. BMC Cancer. 14:7292014. View Article : Google Scholar : PubMed/NCBI
31	Mésange P, Poindessous V, Sabbah M, Escargueil AE, de Gramont A and Larsen AK: Intrinsic bevacizumab resistance is associated with prolonged activation of autocrine VEGF signaling and hypoxia tolerance in colorectal cancer cells and can be overcome by nintedanib, a small molecule angiokinase inhibitor. Oncotarget. 5:4709–4721. 2014. View Article : Google Scholar : PubMed/NCBI
32	Baranasic D, Oppermann T, Cheaib M, Cullum J, Schmidt H and Simon M: Genomic characterization of variable surface antigens reveals a telomere position effect as a prerequisite for RNA interference-mediated silencing in paramecium tetraurelia. MBio. 5:e013282014. View Article : Google Scholar : PubMed/NCBI
33	Luo B, Ju S, Muneri CW and Rui R: Effects of histone acetylation status on the early development of in vitro porcine transgenic cloned embryos. Cell Reprogram. 17:41–48. 2015. View Article : Google Scholar : PubMed/NCBI