With no interaction, knockdown of Apollon and MDR1 reverse the multidrug resistance of human chronic myelogenous leukemia K562/ADM cells

Chen,Jie-Ru; Jia,Xiu-Hong; Wang,Hong; Yi,Ying-Jie; Li,You-Jie

doi:10.3892/or.2017.5535

With no interaction, knockdown of Apollon and MDR1 reverse the multidrug resistance of human chronic myelogenous leukemia K562/ADM cells

Authors:
- Jie-Ru Chen
- Xiu-Hong Jia
- Hong Wang
- Ying-Jie Yi
- You-Jie Li
View Affiliations

Affiliations: Department of Pediatrics, The Affiliated Hospital of Binzhou Medical University, Binzhou, Shandong 256603, P.R. China, Department of Biochemistry and Molecular Biology, Key Laboratory of Tumour Molecular Biology, Binzhou Medical University, Yantai, Shandong 264003, P.R. China
Published online on: March 29, 2017 https://doi.org/10.3892/or.2017.5535
Pages: 2735-2742

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

Chemotherapy is the main treatment method for patients with chronic myeloid leukemia (CML) and has achieved marked results. However, the acquisition of multidrug resistance (MDR) has seriously affected the quality of life and survival rate of patients. The overexpression of the inhibitors of apoptosis proteins (IAPs) and the adenosine triphosphate (ATP)-dependent binding cassette (ABC) transporters are the two main causes of MDR. Apollon and MDR1 are the most important and representative members, respectively, among the IAPs and ABC transporters. In the present study, we investigated the role of Apollon and MDR1 in chemotherapy resistance and their mechanism of interaction. We respectively knocked down the expression of Apollon and MDR1 using short hairpin RNA (shRNA) in adriamycin (ADM) resistant human CML K562 cells and examined the drug sensitivity, the consequences with regard to ADM accumulation and the alterations in the expression of Apollon and MDR1. The expression levels of Apollon and MDR1 mRNA were higher in the K562/ADM cells compared with the parental K562 cells as determined by reverse transcription‑polymerase chain reaction (RT-PCR). The plasmids of Apollon and MDR1 shRNA were respectively stably transfected into K562/ADM cells using Lipofectamine 2000. The transfection efficiency was detected by fluorescence microscopy. Cell Counting Kit-8 (CCK-8) assay revealed that Apollon or MDR1 knockdown significantly increased the chemosensitivity of the K562/ADM cells to ADM. Flow cytometric assay revealed that K562/ADM/shMDR1 cells exhibited a significantly increased intracellular accumulation of ADM, and that changes were not found in the K562/ADM/shApollon cells. Compared with the parental K562/ADM cells, a significantly decreased expression of Apollon mRNA and protein was determined in the K562/ADM/shApollon cells without affecting the expression of MDR1 as determined by RT-PCR and western blotting. Likewise, the expression levels of MDR1 mRNA and protein also markedly downregulated in the K562/ADM/shMDR1 cells had no effect on Apollon expression. Collectively, our findings demonstrated, for the first time, that downregulation of Apollon or MDR1 through stable transfection with the Apollon- or MDR1-targeting shRNA induced MDR reversal through respective inhibition of Apollon or MDR1 expression and function. However, the reversal mechanism of Apollon and MDR1 revealed no direct interaction with each other.

View Figures

View References

1	Apperley JF: Chronic myeloid leukaemia. Lancet. 385:1447–1459. 2015. View Article : Google Scholar : PubMed/NCBI
2	Jabbour E: Chronic myeloid leukemia: First-line drug of choice. Am J Hematol. 91:59–66. 2016. View Article : Google Scholar : PubMed/NCBI
3	Zhang X, Dong W, Zhou H, Li H, Wang N, Miao X and Jia L: α-2,8-Sialyltransferase is involved in the development of multidrug resistance via PI3K/Akt pathway in human chronic myeloid leukemia. IUBMB Life. 67:77–87. 2015. View Article : Google Scholar : PubMed/NCBI
4	Hu M, Liu Y, Deng C, Han R, Jia Y, Liu S, Jiang Z, Cao X, He L and Zhang Q: Enhanced invasiveness in multidrug resistant leukemic cells is associated with overexpression of P-glycoprotein and cellular inhibitor of apoptosis protein. Leuk Lymphoma. 52:1302–1311. 2011. View Article : Google Scholar : PubMed/NCBI
5	Liu F, Liu S, He S, Xie Z, Zu X and Jiang Y: Survivin transcription is associated with P-glycoprotein/MDR1 overexpression in the multidrug resistance of MCF-7 breast cancer cells. Oncol Rep. 23:1469–1475. 2010.PubMed/NCBI
6	Notarbartolo M, Cervello M, Dusonchet L, Cusimano A and D'Alessandro N: Resistance to diverse apoptotic triggers in multidrug resistant HL60 cells and its possible relationship to the expression of P-glycoprotein, Fas and of the novel anti-apoptosis factors IAP (inhibitory of apoptosis proteins). Cancer Lett. 180:91–101. 2002. View Article : Google Scholar : PubMed/NCBI
7	Kachalaki S, Baradaran B, Majidi J, Yousefi M, Shanehbandi D, Mohammadinejad S and Mansoori B: Reversal of chemoresistance with small interference RNA (siRNA) in etoposide resistant acute myeloid leukemia cells (HL-60). Biomed Pharmacother. 75:100–104. 2015. View Article : Google Scholar : PubMed/NCBI
8	Arrigoni E, Galimberti S, Petrini M, Danesi R and Di Paolo A: ATP-binding cassette transmembrane transporters and their epigenetic control in cancer: An overview. Expert Opin Drug Metab Toxicol. 12:1419–1432. 2016. View Article : Google Scholar : PubMed/NCBI
9	Saleem M, Qadir MI, Perveen N and Ahmad B, Saleem U, Irshad T and Ahmad B: Inhibitors of apoptotic proteins: New targets for anticancer therapy. Chem Biol Drug Des. 82:243–251. 2013. View Article : Google Scholar : PubMed/NCBI
10	Notarbartolo M, Cervello M, Poma P, Dusonchet L, Meli M and D'Alessandro N: Expression of the IAPs in multidrug resistant tumor cells. Oncol Rep. 11:133–136. 2004.PubMed/NCBI
11	Zhang S, Tang WQ, Weng SQ, Liu XJ, Rao BQ, Gu JX, Chen S, Wang Q, Shen XZ, Xue RY and Dong L: Apollon modulates chemosensitivity in human esophageal squamous cell carcinoma. Oncotarget. 5:(16). 7183–97. 2014. View Article : Google Scholar : PubMed/NCBI
12	Chen Z, Naito M, Hori S, Mashima T, Yamori T and Tsuruo T: A human IAP-family gene, Apollon, expressed in human brain cancer cells. Biochem Biophys Res Commun. 264:847–854. 1999. View Article : Google Scholar : PubMed/NCBI
13	Peng XX, Tiwari AK, Wu HC and Chen ZS: Overexpression of P-glycoprotein induces acquired resistance to imatinib in chronic myelogenous leukemia cells. Chin J Cancer. 31:110–118. 2012. View Article : Google Scholar : PubMed/NCBI
14	Shi Z, Liang YJ, Chen ZS, Wang XH, Ding Y, Chen LM and Fu LW: Overexpression of Survivin and XIAP in MDR cancer cells unrelated to P-glycoprotein. Oncol Rep. 17:969–976. 2007.PubMed/NCBI
15	Reis FR, Vasconcelos FC, Pereira DL, Moellman-Coelho A, Silva KL and Maia RC: Survivin and P-glycoprotein are associated and highly expressed in late phase chronic myeloid leukemia. Oncol Rep. 26:471–478. 2011.PubMed/NCBI
16	Silva KL, de Souza PS, de Nestal Moraes G, Moellmann-Coelho A, Vasconcelos FC and Maia RC: XIAP and P-glycoprotein co-expression is related to imatinib resistance in chronic myeloid leukemia cells. Leuk Res. 37:1350–1358. 2013. View Article : Google Scholar : PubMed/NCBI
17	Chen JR, Jia XH, Wang H, Yi YJ, Wang JY and Li YJ: Timosaponin A-III reverses multi-drug resistance in human chronic myelogenous leukemia K562/ADM cells via downregulation of MDR1 and MRP1 expression by inhibiting PI3K/Akt signaling pathway. Int J Oncol. 48:2063–2070. 2016.PubMed/NCBI
18	Jia XH, Xiao FF and Li JC: Effect of Apollon siRNA combined with tetramethylpyrazine on proliferation and apoptosis of leukemia K562 cells. Zhongguo Dang Dai Er Ke Za Zhi. 16:135–140. 2014.(In Chinese). PubMed/NCBI
19	Du WT, Liu B, Gu DS, Han ZB, Liu PX, Xu J, Liang L, Zhao HF, Lu SH and Yang RC: Reversal of leukemia multidrug resistance by sequence-specific short hairpin RNA. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 17:563–567. 2009.(In Chinese). PubMed/NCBI
20	Wang L, Meng Q, Wang C, Liu Q, Peng J, Huo X, Sun H, Ma X and Liu K: Dioscin restores the activity of the anticancer agent adriamycin in multidrug-resistant human leukemia K562/adriamycin cells by down-regulating MDR1 via a mechanism involving NF-κB signaling inhibition. J Nat Prod. 76:909–914. 2013. View Article : Google Scholar : PubMed/NCBI
21	Sarkadi B, Homolya L, Szakács G and Váradi A: Human multidrug resistance ABCB and ABCG transporters: Participation in a chemoimmunity defense system. Physiol Rev. 86:1179–1236. 2006. View Article : Google Scholar : PubMed/NCBI
22	Agrawal M, Hanfstein B, Erben P, Wolf D, Ernst T, Fabarius A, Saussele S, Purkayastha D, Woodman RC, Hofmann WK, et al: MDR1 expression predicts outcome of Ph⁺ chronic phase CML patients on second-line nilotinib therapy after imatinib failure. Leukemia. 28:1478–1485. 2014. View Article : Google Scholar : PubMed/NCBI
23	Zu B, Li Y, Wang X, He D, Huang Z and Feng W: MDR1 gene polymorphisms and imatinib response in chronic myeloid leukemia: A meta-analysis. Pharmacogenomics. 15:667–677. 2014. View Article : Google Scholar : PubMed/NCBI
24	Zheng Q, Wu H, Yu Q, Kim DH, Lipton JH, Angelini S, Soverini S, Vivona D, Takahashi N and Cao J: ABCB1 polymorphisms predict imatinib response in chronic myeloid leukemia patients: A systematic review and meta-analysis. Pharmacogenomics J. 15:127–134. 2015. View Article : Google Scholar : PubMed/NCBI
25	Lardo M, Castro M, Moiraghi B, Rojas F, Borda N, Rey JA and Lazarowski A: MDR1/ABCB1 gene polymorphisms in patients with chronic myeloid leukemia. Blood Res. 50:154–159. 2015. View Article : Google Scholar : PubMed/NCBI
26	Sung KW, Choi J, Hwang YK, Lee SJ, Kim HJ, Lee SH, Yoo KH, Jung HL and Koo HH: Overexpression of Apollon, an antiapoptotic protein, is associated with poor prognosis in childhood de novo acute myeloid leukemia. Clin Cancer Res. 13:5109–5114. 2007. View Article : Google Scholar : PubMed/NCBI
27	Ismail EA, Mahmoud HM, Tawfik LM, Habashy DM, Adly AA, El-Sherif NH and Abdelwahab MA: BIRC6/Apollon gene expression in childhood acute leukemia: Impact on therapeutic response and prognosis. Eur J Haematol. 88:118–127. 2012. View Article : Google Scholar : PubMed/NCBI