Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
권호 표지
DOI QR코드

DOI QR Code

RNAi-based Knockdown of Multidrug Resistance-associated Protein 1 is Sufficient to Reverse Multidrug Resistance of Human Lung Cells

  • Shao, Shu-Li (College of Life Sciences and Agriculture and Forestry, Qiqihar University) ;
  • Cui, Ting-Ting (College of Life Sciences and Agriculture and Forestry, Qiqihar University) ;
  • Zhao, Wei (Beijing systerm for Diseases Control and Prevention) ;
  • Zhang, Wei-Wei (College of Life Sciences and Agriculture and Forestry, Qiqihar University) ;
  • Xie, Zhen-Li (College of Life Sciences and Agriculture and Forestry, Qiqihar University) ;
  • Wang, Chang-He (College of Life Sciences and Agriculture and Forestry, Qiqihar University) ;
  • Jia, Hong-Shuang (College of Life Sciences and Agriculture and Forestry, Qiqihar University) ;
  • Liu, Qian (College of Life Sciences and Agriculture and Forestry, Qiqihar University)
  • Published : 2015.01.22
Download PDF
⟨ Previous Next ⟩

Abstract

Up-regulation of multidrug resistance-associated protein 1 (MRP1) is regarded as one of the main causes for multidrug resistance (MDR) of tumor cells, leading to failure of chemotherapy-based treatment for a multitude of cancers. However, whether silencing the overexpressed MRP1 is sufficient to reverse MDR has yet to be validated. This study demonstrated that RNAi-based knockdown of MRP1 reversed the increased efflux ability and MDR efficiently. Two different short haipin RNAs (shRNAs) targeting MRP1 were designed and inserted into pSilence-2.1-neo. The shRNA recombinant plasmids were transfected into cis-dichlorodiamineplatinum-resistant A549 lung (A549/DDP) cells, and then shRNA expressing cell clones were collected and maintained. Real time PCR and immunofluorescence staining for MRP1 revealed a high silent efficiency of these two shRNAs. Functionally, shRNA-expressing cells showed increased rhodamine 123 retention in A549/DDP cells, indicating reduced efflux ability of tumor cells in the absence of MRP1. Consistently, MRP1-silent cells exhibited decreased resistance to 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and DDP, suggesting reversal of MDR in these tumor cells. Specifically, MRP1 knockdown increased the DDP-induced apoptosis of A549/DDP cells by increased trapping of their cell cycling in the G2 stage. Taken together, this study demonstrated that RNAi-based silencing of MRP1 is sufficient to reverse MDR in tumor cells, shedding light on possible novel clinical treatment of cancers.

Keywords

References

  1. Ansaloni S, Lelkes N, Snyder J, et al (2010). A streamlined sub-cloning procedure to transfer shRNA from a pSM2 vector to a pGIPZ lentiviral vector. RNAi Gene Silencing, 296, 550-3.
  2. Chen ZX, Zhu XS, Xie T, et al (2014). Drug resistance reversed by silencing LIM domain-containing protein 1 expression in colorectal carcinoma. Oncol Lett, 8, 795-8.
  3. Chen ZS, Tiwari AK (2011). Multidrug resistance proteins (MRPs/ABCCs) in cancer chemotherapy and genetic diseases. FEBS J, 278, 3226-45. https://doi.org/10.1111/j.1742-4658.2011.08235.x
  4. Cai BL, Liu Y, Xu XF, et al (2013). Nuclear multidrug-resistance related protein 1 contributes to multidrug-resistance of mucoepidermoid carcinoma mainly via regulating multidrug-resistance protein 1: a human mucoepidermoid carcinoma cells model and spearman's rank correlation analysis. PLos One, 9, 69611.
  5. Chen BA, Mao PP, Cheng J, et al (2010). Reversal of multidrug resistance by magnetic Fe3O4 nanoparticle copolymerizating daunorubicin and MDR1 shRNA expression vector in leukemia cells. Int J Nanomedicine, 5, 437-44.
  6. Chen XP, Wang Q, Guan J, et al (2006). Reversing multidrug resistance by RNA interference through the suppression of MDR1 gene in human hepatoma cells. World J Gastroenterol, 12, 3332-7. https://doi.org/10.3748/wjg.v12.i21.3332
  7. Felipe AV, Oliveira J, Chang PY, et al (2014). RNA interference: a promising therapy for gastric cancer. Asian Pac J Cancer Prev, 15, 5509-15. https://doi.org/10.7314/APJCP.2014.15.14.5509
  8. Frouzandeh M, Soodeh A (2012). Multidrug resistance-associated protein 1 predicts relapse in Iranian childhood acute lymphoblastic leukemia. Asian Pac J Cancer Prev, 13, 2285-9. https://doi.org/10.7314/APJCP.2012.13.5.2285
  9. Janet WT, Elizabeth HB, et al (2013). Drug resistance mechanisms in non-small cell lung carcinoma. J Can Res, 2, 265-82.
  10. Ke Li, Baoan Chen, Lin Xu, et al (2013). Reversal of multidrug resistance by cisplatin-loaded magnetic Fe3O4 nanoparticles in A549/DDP lung cancer cells in vitro and in vivo. Int J Nanomedicine, 8, 1867-77.
  11. Kaewpiboon C, Winayanuwattikun P, Yongvanich T, et al (2014). Effect of three fatty acids from the leaf extract of Tiliacora triandra on P-glycoprotein function in multidrug-resistant A549RT-eto cell line. Pharmacogn Mag, 10, 549-56. https://doi.org/10.4103/0973-1296.139779
  12. Kamlesh S, Atish P, Rishil JK, et al (2012). Multidrug resistance associated proteins in multidrug resistance. Chin J Cancer, 31, 58-72. https://doi.org/10.5732/cjc.011.10329
  13. Kepper D (2011 ). Multidrug resistance proteins (MRPs, ABCCs): importance for pathophysiology and drug therapy. Handb Exp Pharmacol, 201, 299-323. https://doi.org/10.1007/978-3-642-14541-4_8
  14. Ketting RF (2011). The many faces of RNAi. Dev Cell, 20, 148-61. https://doi.org/10.1016/j.devcel.2011.01.012
  15. Liang Z, Wu H, Xia J, et al (2010). Involvement of miR-326 in chemotherapy resistance of breast cancer through modulating expression of multidrug resistance-associated protein 1. Biochem Pharmacol, 79, 817-24. https://doi.org/10.1016/j.bcp.2009.10.017
  16. Liang Z, Wu T, Lou H, Yu X, et al (2004). Inhibition of breast cancer metastasis by selective synthetic polypeptide against CXCR4. Cancer Res, 64, 4302-8. https://doi.org/10.1158/0008-5472.CAN-03-3958
  17. Ma H, Cheng L, Hao K, et al (2014). Reversal effect of ST6GAL 1 on multidrug resistance in human leukemia by regulating the PI3K/Akt pathway and the expression of P-gp and MRP1. PLos One, 9, 85113. https://doi.org/10.1371/journal.pone.0085113
  18. Miyagishi M, Taira K (2002). U6 promoter-driven siRNAs with four uridine 3' overhangs efficiently suppress targeted gene expression in mammalian cells. Nat Biotechnol, 20, 497-500. https://doi.org/10.1038/nbt0502-497
  19. Puthucode VR, Savarimuthu I (2012). RNA Interference as a plausible anticancer therapeutic tool. Asian Pac J Cancer Prev, 13, 2445-52. https://doi.org/10.7314/APJCP.2012.13.6.2445
  20. Palaniyandi K, Zhao Q, Chang XB et al (2011). Infection of H69AR cells with retroviral particles harboring interfering RNAi significantly reduced the multidrug resistance of these small cell lung cancer cells. Innt J Biochem Mol Biol, 2, 155-67.
  21. Pecot CV, Calin GA, Coleman RL, et al (2011). RNA interference in the clinic: challenges and future directions. Nat Rev Cancer, 11, 59-67. https://doi.org/10.1038/nrc2966
  22. Pham AN, Wang J, Fang J, et al (2009). Pharmacogenomics approach reveals MRP1 (ABCC1)-mediated resistance to geldanamycins. Harm Res, 26, 936-45.
  23. Sun L, Chen W, Qu L, et al (2013). Icaritin reverses multidrug resistance of HepG2/ADR human hepatoma cells via downregulation of MDR1 and P glycoprotein expressio n. Mol Med Rep, 8, 1883-7.
  24. Shao SL, Sun YY, Li XY, et al (2008). The reversion effect of the RNAi-silencing mdr1 gene on multidrug resistance of the leukemia cell HT9. Cell Biology International, 32, 893-8. https://doi.org/10.1016/j.cellbi.2008.03.021
  25. Shao SL, Zhang WW, Li XY, et al (2010). Reversal of MDR1 gene-dependent multidrugresistance in HL60/HT9 cells using short hairpin RNA expression vectors. Cancer Biotherapy Radiopharmaceuticals, 25, 171-7. https://doi.org/10.1089/cbr.2008.0611
  26. Wu Z, Li XB, Zeng Y, et al (2010). In vitro and in vivo inhibiion of MRP gene expession and reversal of multidrug resistance by siRNA. Basic Clin Pharmacol Toxicol, 108, 177-84.
  27. Wei S, Roessler BC, Chauvet S, et al (2014). Conserved allosteric hot spots in the transmembrane domains of cystic fibrosis transmembrane conductance regulator (CFTR) channels and multidrug resistance protein (MRP) pumps. J Biol Chem, 289, 19942-57. https://doi.org/10.1074/jbc.M114.562116
  28. Xiao H, Wu Z, Shen H, et al (2008). In vivo reversal of P-glycoprotein-mediated multidrug resistance by efficient delivery of stealth RNAi. Basic Clin Pharmacol Toxicol, 103, 342-8. https://doi.org/10.1111/j.1742-7843.2008.00296.x
  29. Ye X, Liu T, Gong Y, et al (2009). Lentivirus-mediated RNA interference reversing the drug-resistance in MDR1 single-factor resistant cell line K562/MDR1. Leu Res, 33, 1114-9. https://doi.org/10.1016/j.leukres.2008.10.011
  30. Yang AK, Zhou ZW, Wei MQ, et al (2010). Modulators of multidrug resistance associated poteins in the management of anticancer and antimicrobial drug resistance and the treatment of inflammatory diseases. Curr Top Med Chem, 10, 1732-56. https://doi.org/10.2174/156802610792928040
  31. Zhu CY, Lv YP , Yan DF, et al (2013). Knockdown of MDR1 increases the sensitivity to adriamycin in drug resistant gastric cancer cells. Asian Pac J Cancer Prev, 14, 6757-60. https://doi.org/10.7314/APJCP.2013.14.11.6757

Cited by

  1. Cisplatin-mediated c-myc overexpression and cytochrome c (cyt c) release result in the up-regulation of the death receptors DR4 and DR5 and the activation of caspase 3 and caspase 9, likely responsible for the TRAIL-sensitizing effect of cisplatin vol.32, pp.4, 2015, https://doi.org/10.1007/s12032-015-0588-9