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Erschienen in: Tumor Biology 2/2016

16.09.2015 | Original Article

Inhibition of microRNA-196a might reverse cisplatin resistance of A549/DDP non-small-cell lung cancer cell line

verfasst von: Jian-Huang Li, Ning Luo, Mei-Zuo Zhong, Zhi-Qiang Xiao, Jian-Xin Wang, Xiao-Yi Yao, Yun Peng, Jun Cao

Erschienen in: Tumor Biology | Ausgabe 2/2016

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Abstract

We aimed to explore the possible mechanism of microRNA-196a (miR-196a) inhibition and reversion of drug resistance to cisplatin (DDP) of the A549/DDP non-small-cell lung cancer (NSCLC) cell line. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect expression differences of miR-196a in the drug-resistant A549/DDP NLCLC cell line and the parental A549 cell line, and expressions of miR-196a in the A549/DDP NLCLC cell line transfected with miR-196a inhibitor (anti-miR-196a group) and the miR-196a negative control (miR-NC) group and blank group (without transfection). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test was applied in examining the cell viability of A549/DDP cell line before and after transfection. Clonogenic assay was used to detect cell proliferation ability. Flow cytometry was applied in detecting apoptosis rate of assayed tumor cell and rhodamine-123 changes in cells. Western blot was applied in detecting proteins of drug-resistant related gene in A549/DDP cell line. Significantly higher expression of miR-196a was detected in the drug-resistant A549/DDP cell line than that in the parental A549 cell line (P < 0.05). However, miR-196a expression in the anti-miR-196a group decreased obviously compared to that in the blank group and the miR-NC group (both P < 0.05); The value of IC50 in the anti-miR-196a group showed remarkably lower than that in the blank group and the miR-NC group (both P < 0.05); Rh-123 absorbing ability in the anti-miR-196a group increased 2.51 times and 2.49 times respectively compared to that in the blank group and the miR-NC group (both P < 0.05). No statistical differences in the apoptosis rate of A549/DDP cell line in the early stage were found among the three groups (all P > 0.05), but the late-stage apoptosis rate in the anti-miR-196a group was significantly higher than that in the blank group and the miR-NC group (both P < 0.05); The expressions of human multidrug resistance 1 (MDR1), multidrug resistance protein 1 (MRP1), excision repair cross-complementation 1 (ERCC1), survivin, and B cell lymphoma 2 (Bcl-2) decreased significantly while RhoE increased significantly in the anti-miR-196a group than the blank group and the miR-NC group (all P < 0.05). Inhibition of miR-196a could reverse cisplatin resistance of A549/DDP cell lines, which might relate with inhibition of drug efflux, down-regulation of drug-resistant protein expression, cell apoptosis, and cell proliferation suppression.
Literatur
1.
Zurück zum Zitat Forde PM, Ettinger DS. Targeted therapy for non-small-cell lung cancer: past, present and future. Expert Rev Anticancer Ther. 2013;13(6):745–58.CrossRefPubMedPubMedCentral Forde PM, Ettinger DS. Targeted therapy for non-small-cell lung cancer: past, present and future. Expert Rev Anticancer Ther. 2013;13(6):745–58.CrossRefPubMedPubMedCentral
2.
Zurück zum Zitat Chen X, Liu Y, Roe OD, Qian Y, Guo R, Zhu L, et al. Gefitinib or erlotinib as maintenance therapy in patients with advanced stage non-small cell lung cancer: a systematic review. PLoS One. 2013;8(3):e59314.CrossRefPubMedPubMedCentral Chen X, Liu Y, Roe OD, Qian Y, Guo R, Zhu L, et al. Gefitinib or erlotinib as maintenance therapy in patients with advanced stage non-small cell lung cancer: a systematic review. PLoS One. 2013;8(3):e59314.CrossRefPubMedPubMedCentral
3.
Zurück zum Zitat Ye MX, Zhao YL, Li Y, Miao Q, Li ZK, Ren XL, et al. Curcumin reverses cis-platin resistance and promotes human lung adenocarcinoma A549/DDP cell apoptosis through hif-1alpha and caspase-3 mechanisms. Phytomedicine. 2012;19(8–9):779–87.CrossRefPubMed Ye MX, Zhao YL, Li Y, Miao Q, Li ZK, Ren XL, et al. Curcumin reverses cis-platin resistance and promotes human lung adenocarcinoma A549/DDP cell apoptosis through hif-1alpha and caspase-3 mechanisms. Phytomedicine. 2012;19(8–9):779–87.CrossRefPubMed
4.
Zurück zum Zitat Brown T, Pilkington G, Bagust A, Boland A, Oyee J, Tudur-Smith C, et al. Clinical effectiveness and cost-effectiveness of first-line chemotherapy for adult patients with locally advanced or metastatic non-small cell lung cancer: a systematic review and economic evaluation. Health Technol Assess. 2013;17(31):1–278.CrossRef Brown T, Pilkington G, Bagust A, Boland A, Oyee J, Tudur-Smith C, et al. Clinical effectiveness and cost-effectiveness of first-line chemotherapy for adult patients with locally advanced or metastatic non-small cell lung cancer: a systematic review and economic evaluation. Health Technol Assess. 2013;17(31):1–278.CrossRef
5.
Zurück zum Zitat Gerber DE, Schiller JH. Maintenance chemotherapy for advanced non-small-cell lung cancer: new life for an old idea. J Clin Oncol. 2013;31(8):1009–20.CrossRefPubMedPubMedCentral Gerber DE, Schiller JH. Maintenance chemotherapy for advanced non-small-cell lung cancer: new life for an old idea. J Clin Oncol. 2013;31(8):1009–20.CrossRefPubMedPubMedCentral
6.
Zurück zum Zitat Mitsudomi T, Morita S, Yatabe Y, Negoro S, Okamoto I, Tsurutani J, et al. West Japan Oncology G. Gefitinib versus cisplatin plus docetaxel in patients with non-small-cell lung cancer harbouring mutations of the epidermal growth factor receptor (wjtog3405): an open label, randomised phase 3 trial. Lancet Oncol. 2010;11(2):121–8.CrossRefPubMed Mitsudomi T, Morita S, Yatabe Y, Negoro S, Okamoto I, Tsurutani J, et al. West Japan Oncology G. Gefitinib versus cisplatin plus docetaxel in patients with non-small-cell lung cancer harbouring mutations of the epidermal growth factor receptor (wjtog3405): an open label, randomised phase 3 trial. Lancet Oncol. 2010;11(2):121–8.CrossRefPubMed
7.
Zurück zum Zitat Oliver TG, Mercer KL, Sayles LC, Burke JR, Mendus D, Lovejoy KS, et al. Chronic cisplatin treatment promotes enhanced damage repair and tumor progression in a mouse model of lung cancer. Genes Dev. 2010;24(8):837–52.CrossRefPubMedPubMedCentral Oliver TG, Mercer KL, Sayles LC, Burke JR, Mendus D, Lovejoy KS, et al. Chronic cisplatin treatment promotes enhanced damage repair and tumor progression in a mouse model of lung cancer. Genes Dev. 2010;24(8):837–52.CrossRefPubMedPubMedCentral
8.
Zurück zum Zitat Wang Y, Chen L, Huang G, He D, He J, Xu W, et al. Klotho sensitizes human lung cancer cell line to cisplatin via pi3k/Akt pathway. PLoS One. 2013;8(2):e57391.CrossRefPubMedPubMedCentral Wang Y, Chen L, Huang G, He D, He J, Xu W, et al. Klotho sensitizes human lung cancer cell line to cisplatin via pi3k/Akt pathway. PLoS One. 2013;8(2):e57391.CrossRefPubMedPubMedCentral
9.
Zurück zum Zitat Cortes-Sempere M, de Miguel MP, Pernia O, Rodriguez C, de Castro CJ, Nistal M, et al. Ibanez de Caceres I. IGFBP-3 methylation-derived deficiency mediates the resistance to cisplatin through the activation of the IGFIR/Akt pathway in non-small cell lung cancer. Oncogene. 2013;32(10):1274–83.CrossRefPubMed Cortes-Sempere M, de Miguel MP, Pernia O, Rodriguez C, de Castro CJ, Nistal M, et al. Ibanez de Caceres I. IGFBP-3 methylation-derived deficiency mediates the resistance to cisplatin through the activation of the IGFIR/Akt pathway in non-small cell lung cancer. Oncogene. 2013;32(10):1274–83.CrossRefPubMed
10.
Zurück zum Zitat Shin DH, Choi YJ, Park JW. SIRT1 and AMPK mediate hypoxia-induced resistance of non-small cell lung cancers to cisplatin and doxorubicin. Cancer Res. 2014;74(1):298–308.CrossRefPubMed Shin DH, Choi YJ, Park JW. SIRT1 and AMPK mediate hypoxia-induced resistance of non-small cell lung cancers to cisplatin and doxorubicin. Cancer Res. 2014;74(1):298–308.CrossRefPubMed
11.
Zurück zum Zitat Das V, Stepankova J, Hajduch M, Miller JH. Role of tumor hypoxia in acquisition of resistance to microtubule-stabilizing drugs. Biochim Biophys Acta. 2015;1855(2):172–82.PubMed Das V, Stepankova J, Hajduch M, Miller JH. Role of tumor hypoxia in acquisition of resistance to microtubule-stabilizing drugs. Biochim Biophys Acta. 2015;1855(2):172–82.PubMed
12.
Zurück zum Zitat Ganesh S, Iyer AK, Weiler J, Morrissey DV, Amiji MM. Combination of siRNA-directed gene silencing with cisplatin reverses drug resistance in human non-small cell lung cancer. Mol Ther Nucleic Acids. 2013;2:e110.CrossRefPubMedPubMedCentral Ganesh S, Iyer AK, Weiler J, Morrissey DV, Amiji MM. Combination of siRNA-directed gene silencing with cisplatin reverses drug resistance in human non-small cell lung cancer. Mol Ther Nucleic Acids. 2013;2:e110.CrossRefPubMedPubMedCentral
13.
Zurück zum Zitat Chen J, Dexheimer TS, Ai Y, Liang Q, Villamil MA, Inglese J, et al. Selective and cell-active inhibitors of the USP1/ UAF1 deubiquitinase complex reverse cisplatin resistance in non-small cell lung cancer cells. Chem Biol. 2011;18(11):1390–400.CrossRefPubMedPubMedCentral Chen J, Dexheimer TS, Ai Y, Liang Q, Villamil MA, Inglese J, et al. Selective and cell-active inhibitors of the USP1/ UAF1 deubiquitinase complex reverse cisplatin resistance in non-small cell lung cancer cells. Chem Biol. 2011;18(11):1390–400.CrossRefPubMedPubMedCentral
14.
Zurück zum Zitat Vinolas N, Provencio M, Reguart N, Cardenal F, Alberola V, Sanchez-Torres JM, et al. Spanish Lung Cancer G. Single nucleotide polymorphisms in MDR1 gen correlates with outcome in advanced non-small-cell lung cancer patients treated with cisplatin plus vinorelbine. Lung Cancer. 2011;71(2):191–8.CrossRefPubMed Vinolas N, Provencio M, Reguart N, Cardenal F, Alberola V, Sanchez-Torres JM, et al. Spanish Lung Cancer G. Single nucleotide polymorphisms in MDR1 gen correlates with outcome in advanced non-small-cell lung cancer patients treated with cisplatin plus vinorelbine. Lung Cancer. 2011;71(2):191–8.CrossRefPubMed
15.
Zurück zum Zitat Du L, Pertsemlidis A. MicroRNA regulation of cell viability and drug sensitivity in lung cancer. Expert Opin Biol Ther. 2012;12(9):1221–39.CrossRefPubMed Du L, Pertsemlidis A. MicroRNA regulation of cell viability and drug sensitivity in lung cancer. Expert Opin Biol Ther. 2012;12(9):1221–39.CrossRefPubMed
16.
Zurück zum Zitat Wang Z, Li Y, Ahmad A, Azmi AS, Kong D, Banerjee S, et al. Targeting miRNAs involved in cancer stem cell and EMT regulation: An emerging concept in overcoming drug resistance. Drug Resist Updat. 2010;13(4–5):109–18.CrossRefPubMedPubMedCentral Wang Z, Li Y, Ahmad A, Azmi AS, Kong D, Banerjee S, et al. Targeting miRNAs involved in cancer stem cell and EMT regulation: An emerging concept in overcoming drug resistance. Drug Resist Updat. 2010;13(4–5):109–18.CrossRefPubMedPubMedCentral
17.
Zurück zum Zitat Giovannetti E, Erozenci A, Smit J, Danesi R, Peters GJ. Molecular mechanisms underlying the role of microRNAs (miRNAs) in anticancer drug resistance and implications for clinical practice. Crit Rev Oncol Hematol. 2012;81(2):103–22.CrossRefPubMed Giovannetti E, Erozenci A, Smit J, Danesi R, Peters GJ. Molecular mechanisms underlying the role of microRNAs (miRNAs) in anticancer drug resistance and implications for clinical practice. Crit Rev Oncol Hematol. 2012;81(2):103–22.CrossRefPubMed
18.
Zurück zum Zitat Xia H, Ooi LL, Hui KM. MicroRNA-216a/217-induced epithelial-mesenchymal transition targets PTEN and SMAD7 to promote drug resistance and recurrence of liver cancer. Hepatology. 2013;58(2):629–41.CrossRefPubMed Xia H, Ooi LL, Hui KM. MicroRNA-216a/217-induced epithelial-mesenchymal transition targets PTEN and SMAD7 to promote drug resistance and recurrence of liver cancer. Hepatology. 2013;58(2):629–41.CrossRefPubMed
19.
Zurück zum Zitat Fu X, Tian J, Zhang L, Chen Y, Hao Q. Involvement of microRNA-93, a new regulator of PTEN/Akt signaling pathway, in regulation of chemotherapeutic drug cisplatin chemosensitivity in ovarian cancer cells. FEBS Lett. 2012;586(9):1279–86.CrossRefPubMed Fu X, Tian J, Zhang L, Chen Y, Hao Q. Involvement of microRNA-93, a new regulator of PTEN/Akt signaling pathway, in regulation of chemotherapeutic drug cisplatin chemosensitivity in ovarian cancer cells. FEBS Lett. 2012;586(9):1279–86.CrossRefPubMed
20.
21.
Zurück zum Zitat Sun M, Liu XH, Li JH, Yang JS, Zhang EB, Yin DD, et al. MiR-196a is upregulated in gastric cancer and promotes cell proliferation by downregulating p27(KIP1). Mol Cancer Ther. 2012;11(4):842–52.CrossRefPubMed Sun M, Liu XH, Li JH, Yang JS, Zhang EB, Yin DD, et al. MiR-196a is upregulated in gastric cancer and promotes cell proliferation by downregulating p27(KIP1). Mol Cancer Ther. 2012;11(4):842–52.CrossRefPubMed
22.
Zurück zum Zitat Liu XH, Lu KH, Wang KM, Sun M, Zhang EB, Yang JS, et al. MicroRNA-196a promotes non-small cell lung cancer cell proliferation and invasion through targeting HOXA5. BMC Cancer. 2012;12:348.CrossRefPubMedPubMedCentral Liu XH, Lu KH, Wang KM, Sun M, Zhang EB, Yang JS, et al. MicroRNA-196a promotes non-small cell lung cancer cell proliferation and invasion through targeting HOXA5. BMC Cancer. 2012;12:348.CrossRefPubMedPubMedCentral
23.
Zurück zum Zitat Schimanski CC, Frerichs K, Rahman F, Berger M, Lang H, Galle PR, et al. High miR-196a levels promote the oncogenic phenotype of colorectal cancer cells. World J Gastroenterol. 2009;15(17):2089–96.CrossRefPubMedPubMedCentral Schimanski CC, Frerichs K, Rahman F, Berger M, Lang H, Galle PR, et al. High miR-196a levels promote the oncogenic phenotype of colorectal cancer cells. World J Gastroenterol. 2009;15(17):2089–96.CrossRefPubMedPubMedCentral
24.
Zurück zum Zitat Hoffman AE, Zheng T, Yi C, Leaderer D, Weidhaas J, Slack F, et al. MicroRNA miR-196a-2 and breast cancer: a genetic and epigenetic association study and functional analysis. Cancer Res. 2009;69(14):5970–7.CrossRefPubMedPubMedCentral Hoffman AE, Zheng T, Yi C, Leaderer D, Weidhaas J, Slack F, et al. MicroRNA miR-196a-2 and breast cancer: a genetic and epigenetic association study and functional analysis. Cancer Res. 2009;69(14):5970–7.CrossRefPubMedPubMedCentral
25.
Zurück zum Zitat Kim YJ, Bae SW, Yu SS, Bae YC, Jung JS. MiR-196a regulates proliferation and osteogenic differentiation in mesenchymal stem cells derived from human adipose tissue. J Bone Miner Res. 2009;24(5):816–25.CrossRefPubMed Kim YJ, Bae SW, Yu SS, Bae YC, Jung JS. MiR-196a regulates proliferation and osteogenic differentiation in mesenchymal stem cells derived from human adipose tissue. J Bone Miner Res. 2009;24(5):816–25.CrossRefPubMed
26.
Zurück zum Zitat Zhang J, Zhang T, Ti X, Shi J, Wu C, Ren X, et al. Curcumin promotes apoptosis in A549/DDP multidrug-resistant human lung adenocarcinoma cells through an miRNA signaling pathway. Biochem Biophys Res Commun. 2010;399(1):1–6.CrossRefPubMed Zhang J, Zhang T, Ti X, Shi J, Wu C, Ren X, et al. Curcumin promotes apoptosis in A549/DDP multidrug-resistant human lung adenocarcinoma cells through an miRNA signaling pathway. Biochem Biophys Res Commun. 2010;399(1):1–6.CrossRefPubMed
27.
Zurück zum Zitat Olaussen KA, Dunant A, Fouret P, Brambilla E, Andre F, Haddad V, et al. DNA repair by ERCC1 in non-small-cell lung cancer and cisplatin-based adjuvant chemotherapy. N Engl J Med. 2006;355(10):983–91.CrossRefPubMed Olaussen KA, Dunant A, Fouret P, Brambilla E, Andre F, Haddad V, et al. DNA repair by ERCC1 in non-small-cell lung cancer and cisplatin-based adjuvant chemotherapy. N Engl J Med. 2006;355(10):983–91.CrossRefPubMed
28.
Zurück zum Zitat Wang S, Huang X, Lee CK, Liu B. Elevated expression of ERBB3 confers paclitaxel resistance in ERBB2-overexpressing breast cancer cells via upregulation of survivin. Oncogene. 2010;29(29):4225–36.CrossRefPubMed Wang S, Huang X, Lee CK, Liu B. Elevated expression of ERBB3 confers paclitaxel resistance in ERBB2-overexpressing breast cancer cells via upregulation of survivin. Oncogene. 2010;29(29):4225–36.CrossRefPubMed
29.
Zurück zum Zitat Grise F, Sena S, Bidaud-Meynard A, Baud J, Hiriart JB, Makki K, et al. Rnd3/RhoE is down-regulated in hepatocellular carcinoma and controls cellular invasion. Hepatology. 2012;55(6):1766–75.CrossRefPubMed Grise F, Sena S, Bidaud-Meynard A, Baud J, Hiriart JB, Makki K, et al. Rnd3/RhoE is down-regulated in hepatocellular carcinoma and controls cellular invasion. Hepatology. 2012;55(6):1766–75.CrossRefPubMed
30.
Zurück zum Zitat Zhao X, Yang L, Hu J, Ruan J. MiR-138 might reverse multidrug resistance of leukemia cells. Leuk Res. 2010;34(8):1078–82.CrossRefPubMed Zhao X, Yang L, Hu J, Ruan J. MiR-138 might reverse multidrug resistance of leukemia cells. Leuk Res. 2010;34(8):1078–82.CrossRefPubMed
31.
Zurück zum Zitat Wang Q, Zhong M, Liu W, Li J, Huang J, Zheng L. Alterations of microRNAs in cisplatin-resistant human non-small cell lung cancer cells (A549/DDP). Exp Lung Res. 2011;37(7):427–34.CrossRefPubMed Wang Q, Zhong M, Liu W, Li J, Huang J, Zheng L. Alterations of microRNAs in cisplatin-resistant human non-small cell lung cancer cells (A549/DDP). Exp Lung Res. 2011;37(7):427–34.CrossRefPubMed
Metadaten
Titel
Inhibition of microRNA-196a might reverse cisplatin resistance of A549/DDP non-small-cell lung cancer cell line
verfasst von
Jian-Huang Li
Ning Luo
Mei-Zuo Zhong
Zhi-Qiang Xiao
Jian-Xin Wang
Xiao-Yi Yao
Yun Peng
Jun Cao
Publikationsdatum
16.09.2015
Verlag
Springer Netherlands
Erschienen in
Tumor Biology / Ausgabe 2/2016
Print ISSN: 1010-4283
Elektronische ISSN: 1423-0380
DOI
https://doi.org/10.1007/s13277-015-4017-7

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