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

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

doi:10.3892/ijo.2016.3423

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

Authors:
- Jie-Ru Chen
- Xiu-Hong Jia
- Hong Wang
- Ying-Jie Yi
- Jian-Yong Wang
- 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 7, 2016 https://doi.org/10.3892/ijo.2016.3423
Pages: 2063-2070

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

One of the major causes of failure in chemotherapy for patients with human chronic myelogenous leukemia (CML) is the acquisition of multidrug resistance (MDR). MDR is often associated with the overexpression of drug efflux transporters of the ATP-binding cassette (ABC) protein family. Timosaponin A-III (TAIII), a saponin isolated from the rhizome of Anemarrhena asphodeloides, has previously demonstrated the ability to suppress certain human tumor processes and the potential to be developed as an anticancer agent. Nevertheless, the ability of TAIII to reverse MDR has not yet been explored. In this study, the adriamycin (ADM) resistance reversal effect of TAIII in human CML K562/ADM cells and the underlying mechanism was investigated. The Cell Counting Kit-8 (CCK-8) assay showed that TAIII had a reversal effect on the drug resistance of K562/ADM cells. Flow cytometry assay showed increased intracellular accumulation of ADM after cells were pretreated with TAIII, and the changes in the accumulation of rhodamine-123 (Rho-123) and 5(6)-carboxyfluorescein diacetate (CFDA) dye in K562/ADM cells were determined to be similar to the changes of intracellular accumulation of ADM. After pretreatment of cells with TAIII, the decreasing expression of P-gp and MRP1 mRNA was examined by reverse transcription polymerase chain reaction (RT-PCR). Western blotting showed TAIII inhibiting P-gp and MRP1 expression depended on the PI3K/Akt signaling pathway by decreasing the activity of p-Akt. Moreover, wortmannin an inhibitor of PI3K/Akt signaling pathway has a strong inhibitory effect on the expression of p-Akt, P-gp and MRP1. Besides, the combined treatment with TAIII did not have an affect on wortmannin downregulation of p-Akt, P-gp and MRP1. Taken together, our findings demonstrate, for the first time, that TAIII induced MDR reversal through inhibition of P-gp and MRP1 expression and function with regained adriamycin sensitivity which might mainly correlate to the regulation of PI3K/Akt signaling pathway.

View Figures

View References

1	Rodrigues AS, Dinis J, Gromicho M, Martins C, Laires A and Rueff J: Genomics and cancer drug resistance. Curr Pharm Biotechnol. 13:651–673. 2012. View Article : Google Scholar
2	Zhang X, Dong W, Zhou H, Li H, Wang N, Miao X and Jia L: a-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
3	Kalle AM, Sachchidanand S and Pallu R: Bcr-Abl-independent mechanism of resistance to imatinib in K562 cells: Induction of cyclooxygenase-2 (COX-2) by histone deacetylases (HDACs). Leuk Res. 34:1132–1138. 2010. View Article : Google Scholar : PubMed/NCBI
4	Ma H, Cheng L, Hao K, Li Y, Song X, Zhou H and Jia L: 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:e851132014. View Article : Google Scholar : PubMed/NCBI
5	Cheng Z, Yang N, Liang W, Yan X, Li L and Pan L: Effect of PTEN gene transfection on the reversal of multidrug resistance in K562/ADM cells. Leuk Lymphoma. 53:1383–1389. 2012. View Article : Google Scholar
6	Dharmapuri G, Doneti R, Philip GH and Kalle AM: Celecoxib sensitizes imatinib-resistant K562 cells to imatinib by inhibiting MRP1–5, ABCA2 and ABCG2 transporters via Wnt and Ras signaling pathways. Leuk Res. 39:696–701. 2015. View Article : Google Scholar : PubMed/NCBI
7	Kang Q and Yan S: Piperlongumine reverses doxorubicin resistance through the PI3K/Akt signaling pathway in K562/A02 human leukemia cells. Exp Ther Med. 9:1345–1350. 2015.PubMed/NCBI
8	Zhang Z, Xu K, Bi Y, Yu G, Wang S, Qi X and Zhong H: Low intensity ultrasound promotes the sensitivity of rat brain glioma to doxorubicin by downregulating the expressions of P-glucoprotein and multidrug resistance protein 1 in vitro and in vivo. PLoS One. 8:e706852013. View Article : Google Scholar
9	Barrett D, Brown VI, Grupp SA and Teachey DT: Targeting the PI3K/AKT/mTOR signaling axis in children with hematologic malignancies. Paediatr Drugs. 14:299–316. 2012.PubMed/NCBI
10	Sheng Z, Ma L, Sun JE, Zhu LJ and Green MR: BCR-ABL suppresses autophagy through ATF5-mediated regulation of mTOR transcription. Blood. 118:2840–2848. 2011. View Article : Google Scholar : PubMed/NCBI
11	Li Q, Wu Y, Fang S, Wang L, Qi H, Zhang Y, Zhang J and Li W: BCR/ABL oncogene-induced PI3K signaling pathway leads to chronic myeloid leukemia pathogenesis by impairing immunomodulatory function of hemangioblasts. Cancer Gene Ther. 22:227–237. 2015. View Article : Google Scholar : PubMed/NCBI
12	Wang Z, Yang L, Xia Y, Guo C and Kong L: Icariin enhances cytotoxicity of doxorubicin in human multidrug-resistant osteosarcoma cells by inhibition of ABCB1 and down-regulation of the PI3K/Akt pathway. Biol Pharm Bull. 38:277–284. 2015. View Article : Google Scholar : PubMed/NCBI
13	Xie X, Tang B, Zhou J, Gao Q and Zhang P: Inhibition of the PI3K/Akt pathway increases the chemosensitivity of gastric cancer to vincristine. Oncol Rep. 30:773–782. 2013.PubMed/NCBI
14	Tsou SH, Chen TM, Hsiao HT and Chen YH: A critical dose of doxorubicin is required to alter the gene expression profiles in MCF-7 cells acquiring multidrug resistance. PLoS One. 10:e01167472015. View Article : Google Scholar : PubMed/NCBI
15	Cheng L, Luo S, Jin C, Ma H, Zhou H and Jia L: FUT family mediates the multidrug resistance of human hepatocellular carcinoma via the PI3K/Akt signaling pathway. Cell Death Dis. 4:e9232013. View Article : Google Scholar : PubMed/NCBI
16	Lee B, Jung K and Kim DH: Timosaponin AIII, a saponin isolated from Anemarrhena asphodeloides, ameliorates learning and memory deficits in mice. Pharmacol Biochem Behav. 93:121–127. 2009. View Article : Google Scholar : PubMed/NCBI
17	Lim SM, Jeong JJ, Kang GD, Kim KA, Choi HS and Kim DH: Timosaponin AIII and its metabolite sarsasapogenin ameliorate colitis in mice by inhibiting NF-κB and MAPK activation and restoring Th17/Treg cell balance. Int Immunopharmacol. 25:493–503. 2015. View Article : Google Scholar : PubMed/NCBI
18	Lee B, Trinh HT, Jung K, Han SJ and Kim DH: Inhibitory effects of steroidal timosaponins isolated from the rhizomes of Anemarrhena asphodeloides against passive cutaneous anaphylaxis reaction and pruritus. Immunopharmacol Immunotoxicol. 32:357–363. 2010. View Article : Google Scholar : PubMed/NCBI
19	Kimura M, Kimura I and Chem FJ: Combined potentiating effect of byakko-ka-ninjin-to, its constituents, rhizomes of Anemarrhena asphodeloitles, timosaponin A-III, and calcium on pilocarpine-induced saliya secretion in streptozocin-diabetic mice. Biol Pharm Bull. 19:926–931. 1996. View Article : Google Scholar : PubMed/NCBI
20	Tang YH, Sun ZL, Fan MS, Li ZX and Huang CG: Anti-diabetic effects of TongGuanWan, a Chinese traditional herbal formula, in C57BL/KsJ-db/db mice. Planta Med. 78:18–23. 2012. View Article : Google Scholar
21	Lok CN, Sy LK, Liu F and Che CM: Activation of autophagy of aggregation-prone ubiquitinated proteins by timosaponin A-III. J Biol Chem. 286:31684–31696. 2011. View Article : Google Scholar : PubMed/NCBI
22	Kang YJ, Chung HJ, Nam JW, Park HJ, Seo EK, Kim YS, Lee D and Lee SK: Cytotoxic and antineoplastic activity of timosaponin A-III for human colon cancer cells. J Nat Prod. 74:701–706. 2011. View Article : Google Scholar : PubMed/NCBI
23	Wang YZ, Feng B, Huang HZ, Kang LP, Cong Y, Zhou WB, Zou P, Cong YW, Song XB and Ma BP: Glucosylation of steroidal saponins by cyclodextrin glucanotransferase. Planta Med. 76:1724–1731. 2010. View Article : Google Scholar : PubMed/NCBI
24	Huang HL, Chiang WL, Hsiao PC, Chien MH, Chen HY, Weng WC, Hsieh MJ and Yang SF: Timosaponin AIII mediates caspase activation and induces apoptosis through JNK1/2 pathway in human promyelocytic leukemia cells. Tumour Biol. 36:3489–3497. 2015. View Article : Google Scholar
25	Wang S, Wang S, Wang C, Chen Y, Li J, Wang X, Wang D, Li Z, Peng Z and Fan L: Upregulation of multidrug resistance-associated protein 1 by Allyl isothiocyanate in human bronchial epithelial cell: Involvement of c-Jun N-terminal kinase signaling pathway. Oxid Med Cell. 2015:9037822015.
26	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
27	Wang H, Wang X, Li Y, Liao A, Fu B, Pan H, Liu Z and Yang W: The proteasome inhibitor bortezomib reverses P-glycoprotein-mediated leukemia multi-drug resistance through the NF-B pathway. Pharmazie. 67:187–192. 2012.PubMed/NCBI
28	Xu X, Zhang Y, Li W, Miao H, Zhang H, Zhou Y, Li Z, You Q, Zhao L and Guo Q: Wogonin reverses multi-drug resistance of human myelogenous leukemia K562/A02 cells via downregulation of MRP1 expression by inhibiting Nrf2/ARE signaling pathway. Biochem Pharmacol. 92:220–234. 2014. View Article : Google Scholar : PubMed/NCBI
29	Sinclair A, Latif AL and Holyoake TL: Targeting survival pathways in chronic myeloid leukaemia stem cells. Br J Pharmacol. 169:1693–1707. 2013. View Article : Google Scholar : PubMed/NCBI
30	Lage H: An overview of cancer multidrug resistance: A still un-solved problem. Cell Mol Life Sci. 65:3145–3167. 2008. View Article : Google Scholar : PubMed/NCBI
31	Vasconcelos FC, Silva KL, Souza PS, Silva LF, Moellmann-Coelho A, Klumb CE and Maia RC: Variation of MDR proteins expression and activity levels according to clinical status and evolution of CML patients. Cytometry B Clin Cytom. 80B:158–166. 2011. View Article : Google Scholar
32	Sy LK, Yan SC, Lok CN, Man RY and Che CM: Timosaponin A-III induces autophagy preceding mitochondria-mediated apoptosis in HeLa cancer cells. Cancer Res. 68:10229–10237. 2008. View Article : Google Scholar : PubMed/NCBI
33	Wang N, Feng Y, Zhu M, Siu FM, Ng KM and Che CM: A novel mechanism of XIAP degradation induced by timosaponin AIII in hepatocellular carcinoma. Biochim Biophys Acta. 1833:2890–2899. 2013. View Article : Google Scholar : PubMed/NCBI
34	King FW, Fong S, Griffin C, Shoemaker M, Staub R, Zhang YL, Cohen I and Shtivelman E: Timosaponin AIII is preferentially cytotoxic to tumor cells through inhibition of mTOR and induction of ER stress. PLoS One. 4:e72832009. View Article : Google Scholar : PubMed/NCBI
35	Gao F, Liu J, Dong WW, Wang W, Wang Y, Cai D, Zheng Z and Sun K: Investigation of the mechanism involved in the As₂O₃-regulated decrease in MDR1 expression in leukemia cells. Oncol Rep. 31:926–932. 2014.
36	Wang R, Shao F, Liu Z, Zhang J, Wang S, Liu J, Liu H, Chen H, Liu K, Xia M, et al: The Hsp90 inhibitor SNX-2112, induces apoptosis in multidrug resistant K562/ADR cells through suppression of Akt/NF-κB and disruption of mitochondria-dependent pathways. Chem Biol Interact. 205:1–10. 2013. View Article : Google Scholar : PubMed/NCBI
37	Zhu X, Wang L, Zhang B, Li J, Dou X and Zhao RC: TGF-beta1-induced PI3K/Akt/NF-kappaB/MMP9 signalling pathway is activated in Philadelphia chromosome-positive chronic myeloid leukaemia hemangioblasts. J Biochem. 149:405–414. 2011. View Article : Google Scholar : PubMed/NCBI