Advertisement

Novel combination of tanshinone I and lenalidomide induces chemo-sensitivity in myeloma cells by modulating telomerase activity and expression of shelterin complex and its associated molecules

  • Raman Kumar
  • Nidhi Gupta
  • Himani
  • Alpana Sharma
Original Article

Abstract

Shelterin complex and its associated molecules are imperative for proper functioning and maintenance of human telomeres. These molecules in association with human telomerase have been found altered in most cancers including multiple myeloma thereby proposed them as suitable therapeutic targets. Further, due to aggressive and recurring behavior of myeloma novel, efficacious and safe therapeutic agents for disease prevention are primary requirements for treatment of this disease. This maiden attempt evaluated the anti-proliferative properties of tanshinone I (TanI) alone or in combination with lenalidomide (Len) on myeloma cancer cell lines (RPMI8226 and U226). Further, after drug treatment levels of telomerase activity (TA) and molecular expression (mRNA & protein) of shelterin complex and its associated molecules have also been investigated. Results demonstrated that, TanI significantly inhibited proliferation of myeloma cells in dose and time dependent manner as observed through cytotoxicity assay. Additionally, induction of apoptosis by TanI and in combination with Len was observed in myeloma cells through propidium iodide (PI) staining, annexin V-FITC/PI staining, TUNEL and caspase-3/7 activity assays. Further, drug treatment significantly decreased (p < 0.01) TA and molecular expression of ACD, TERF2IP and TANK1 in comparison to vehicle control (0.1% DMSO) myeloma cells. Thus, this maiden in-vitro study provided initial evidences of therapeutic potential of TanI alone or in combination with chemotherapeutic agent Len as novel anticancer agents in myeloma cells which need further evaluation in future. Lastly, down-regulation of TA and decreased expression of these molecules underscores their potential as plausible therapeutic targets.

Keywords

Myeloma Tanshinone I Lenalidomide Apoptosis Telomerase Shelterin complex 

Notes

Acknowledgements

Financial assistance to Mr. Raman Kumar as senior research fellow by Indian Council of Medical Research (ICMR), New Delhi, India is acknowledged.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants and/or animals

This article does not contain any study with human participants or animals performed by any of the authors.

References

  1. 1.
    Munshi NC, Anderson KC (2013) New strategies in the treatment of multiple myeloma. Clin Cancer Res 19:3337–3344.  https://doi.org/10.1158/1078-0432.CCR-12-188 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Bianchi G, Richardson PG, Anderson KC (2015) Promising therapies in multiple myeloma. Blood 126:300–310.  https://doi.org/10.1182/blood-2015-03-575365 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Kumar SK, Lee JH, Lahuerta JJ, Morgan G, Richardson PG, Crowley J, Haessler J, Feather J, Hoering A, Moreau P, LeLeu X (2012) Risk of progression and survival in multiple myeloma relapsing after therapy with IMiDs and bortezomib: a multicenter international myeloma working group study. Leukemia 26:149–157.  https://doi.org/10.1038/leu.2011.196 CrossRefPubMedGoogle Scholar
  4. 4.
    Day JP, Marder BA, Morgan WF (1993) Telomeres and their possible role in chromosome stabilization. Environ Mol Mutagen 22:245–249.  https://doi.org/10.1002/em.2850220411 CrossRefPubMedGoogle Scholar
  5. 5.
    Blackburn EH, Greider CW, Henderson E, Lee MS, Shampay J, Lentzen DS (1989) Recognition and elongation of telomeres by telomerase. Genome 31:553–560.  https://doi.org/10.1139/g89-104 CrossRefPubMedGoogle Scholar
  6. 6.
    Shay JW (1997) Telomerase in human development and cancer. J Cell Physiol 173:266–270.  https://doi.org/10.1002/(SICI)1097-4652(199711)173:2%3C266::AID-JCP33%3E3.0.CO;2-B CrossRefPubMedGoogle Scholar
  7. 7.
    Kim NW, Piatyszek MA, Prowse KR, Harley CB, West MD, Ho PL, Coviello GM, Wright WE, Weinrich SL, Shay JW (1994) Specific association of human telomerase activity with immortal cells and cancer. Science 266:2011–2015.  https://doi.org/10.1126/science.7605428 CrossRefGoogle Scholar
  8. 8.
    Park HS, Choi J, See CJ, Kim JA, Park SN, Im MSK, Kim SM, Lee DS, Hwang SM (2017) Dysregulation of telomere lengths and telomerase activity in myelodysplastic syndrome. Ann Lab Med 37:195–203.  https://doi.org/10.3343/alm.2017.37.3.195 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Miyazaki Y, Yoshida N, Nozaki T, Inoue H, Kikuchi K, Kusamaet K (2015) Telomerase activity in the occurrence and progression of oral squamous cell carcinoma. J Oral Sci 57:295–303.  https://doi.org/10.2334/josnusd.57.295 CrossRefPubMedGoogle Scholar
  10. 10.
    Kumar R, Khan R, Gupta N, Seth T, Sharma A, Kalaivani M, Sharma A (2018) Identifying the biomarker potential of telomerase activity and shelterin complex molecule, telomeric repeat binding factor 2 (TERF2), in multiple myeloma. Leuk Lymphoma 59:1677–1689.  https://doi.org/10.1080/10428194.2017.1387915 CrossRefPubMedGoogle Scholar
  11. 11.
    De Lange T (2005) Shelterin: the protein complex that shapes and safeguards human telomeres. Gene Dev 19:2100–2110.  https://doi.org/10.1101/gad.1346005 CrossRefPubMedGoogle Scholar
  12. 12.
    Zhong FL, Batista LFZ, Freund A, Pech MF, Venteicher AS, Artandi SE (2012) TPP1 OB-fold domain controls telomere maintenance by recruiting telomerase to chromosome ends. Cell 150:481–494.  https://doi.org/10.1016/j.cell.2012.07.012 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Tang T, Zhou FX, Lei H, Yu HJ, Xie CH, Zhou YF, Liu SQ (2009) Increased expression of telomere related proteins correlates with resistance to radiation in human laryngeal cancer cell lines. Oncol Rep 21:1505–1509.  https://doi.org/10.3892/or00000381 CrossRefPubMedGoogle Scholar
  14. 14.
    Yang L, Wang W, Hu L, Yang X, Zhong J, Li Z, Yand H, Lei H, Yu H, Liao ZK, Zhou F, Xie C, Zhou Y (2013) Telomere-binding protein TPP1 modulates telomere homeostasis and confers radio resistance to human colorectal cancer cells. PLoS ONE 8:e81034.  https://doi.org/10.1371/journal.pone.0081034 CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Li X, Liu W, Wang H, Yang L, Li Y, Wen H, Ning H, Wang J, Zhang L, Li J, Fan D (2015) Rap1 is indispensable for TRF2 function in etoposide-induced DNA damage response in gastric cancer cell line. Oncogenesis 4:e144, 1–5.  https://doi.org/10.1038/oncsis.2015.1 CrossRefGoogle Scholar
  16. 16.
    Lu H, Lei Z, Lu Z, Lu Q, Lu C, Chen W, Wang C, Tang Q, Kong Q (2013) Silencing tankyrase and telomerase promotes A549 human lung adenocarcinoma cell apoptosis and inhibits proliferation. Oncol Rep 30:1745–1752.  https://doi.org/10.3892/or.2013.2665 CrossRefPubMedGoogle Scholar
  17. 17.
    Wang S, Zhang H, Zhu J, Li C, Zhu J, Bowen S, Zhang B, Wang C (2017) PinX1 is a potential prognostic factor for non-small-cell lung cancer and inhibits cell proliferation and migration. Biomed Res Int 2017:1–9.  https://doi.org/10.1155/2017/7956437 CrossRefGoogle Scholar
  18. 18.
    Turner JG, Dawson J, Emmons MF, Cubitt CL, Kauffman M, Shacham S, Hazlejurst LA, Sullivan DM (2013) CRM1 inhibition sensitizes drug resistant human myeloma cells to topoisomerase II and proteasome inhibitors both in vitro and ex vivo. J Cancer 4:614–625.  https://doi.org/10.7150/jca.7080 CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Nooka AK, Kastritis E, Dimopoulos MA, Lonial S (2015) Treatment options for relapsed and refractory multiple myeloma. Blood 125:3085–3099.  https://doi.org/10.1182/blood-2014-11-568923 CrossRefPubMedGoogle Scholar
  20. 20.
    Normile D (2003) The new face of traditional Chinese medicine. Science 299:188–190.  https://doi.org/10.1126/science.299.5604.188 CrossRefPubMedGoogle Scholar
  21. 21.
    Wang M, Cao J, Zhu JY, Qiu J, Zhang Y, Shu B, Ou TM, Tan JH, Gu LQ, Huang ZS, Sheng Y, Li D (2017) Curcusone C induces telomeric DNA-damage response in cancer cells through inhibition of telomeric repeat factor 2. BBA 1865:1372–1382.  https://doi.org/10.1016/j.bbapap.2017.08.022 CrossRefGoogle Scholar
  22. 22.
    Li Y, Gong Y, Li L, Abdolmaleky HM, Zhou JR (2013) Bioactive tanshinone I inhibits the growth of lung cancer in part via downregulation of Aurora A function. Mol Carcinogen 52:535–543.  https://doi.org/10.1002/mc.21888 CrossRefGoogle Scholar
  23. 23.
    Lee CY, Sher HF, Chen HW, Liu CC, Chen CH, Lin CS, Yang PC, Tsay HS, Chen JJ (2008) Anticancer effects of tanshinone I in human non-small cell lung cancer. Mol Cancer Ther 7:3527–3538.  https://doi.org/10.1158/1535-7163.MCT-07-2288 CrossRefPubMedGoogle Scholar
  24. 24.
    Liu XD, Fan RF, Zhang Y, Yang HZ, Fang ZG, Guan WB, Lin DJ, Xiao RZ, Huang RW, Huang HQ, Liu PQ (2010) Down-regulation of telomerase activity and activation of caspase-3 are responsible for tanshinone I-induced apoptosis in monocyte leukemia cells in vitro. Int J Mol Sci 11:2267–2280.  https://doi.org/10.3390/ijms11062267 CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Soares J, Keppler BR, Wang X, Lee KH, Jarstfer MB (2011) Ortho-Quinone tanshinones directly inhibit telomerase through an oxidative mechanism mediated by hydrogen peroxide. Bioorg Med Chem Lett 21:7474–7478.  https://doi.org/10.1016/j.bmcl.2011.09.112 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254.  https://doi.org/10.1016/0003-2697(76)90527-3 CrossRefGoogle Scholar
  27. 27.
    Liu JJ, Zhang Y, Lin DJ, Xiao RZ (2009) Tanshinone IIA inhibits leukemia THP-1 cell growth by induction of apoptosis. Oncol Rep 21:1075–1081.  https://doi.org/10.3892/or_00000326 CrossRefPubMedGoogle Scholar
  28. 28.
    Shin EA, Sohn EJ, Won G, Choi JU, Jeong M, Kim B, Kim MJ, Kim SH (2014) Upregulation of microRNA135a-3p and death receptor 5 plays a critical role in Tanshinone I sensitized prostate cancer cells to TRAIL induced apoptosis. Oncotarget 5:5624–5636.  https://doi.org/10.18632/oncotarget.2152 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Bashash D, Delshad M, Safaroghli-Azar A, Safa M, Momeny M, Ghaffari SH (2017) Novel pan PI3K inhibitor-induced apoptosis in APL cells correlates with suppression of telomerase: an emerging mechanism of action of BKM120. Int J Biochem Cell Biol 91:1–8.  https://doi.org/10.1016/j.biocel.2017.08.009 CrossRefPubMedGoogle Scholar
  30. 30.
    Weiss C, Uziel O, Wolach O, Nordenberg J, Beery E, Bulvick S, Kanfer G, Cohen O, Ram R, Bakhanashvili M, Magen-Nativ H (2012) Differential downregulation of telomerase activity by bortezomib in multiple myeloma cells-multiple regulatory pathways in vitro and ex vivo. Br J Cancer 107:1844–1852.  https://doi.org/10.1038/bjc.2012.460 CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Shammas MA, Koley H, Bertheau RC, Neri P, Fulciniti M, Tassone P, Blotta S, Protopopov A, Mitsiades C, Batchu RB, Anderson KC (2008) Telomerase inhibitor GRN163L inhibits myeloma cell growth in vitro and in vivo. Leukemia 22:1410–1418.  https://doi.org/10.1038/leu.2008.81 CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Song YYSL, Yuan SL, Yang YM, Wang XJ, Huang GQ (2005) Alteration of activities of telomerase in tanshinone IIA inducing apoptosis of the leukemia cells. Zhongguo Zhong Yao Za Zhi 30:207–211PubMedGoogle Scholar
  33. 33.
    Quach H, Ritchie D, Stewart AK, Neeson P, Harrison S, Smyth MJ, Prince HM (2010) Mechanism of action of immunomodulatory drugs (IMiDS) in multiple myeloma. Leukemia 24:22–32.  https://doi.org/10.1038/leu.2009.236 CrossRefPubMedGoogle Scholar
  34. 34.
    Jian Y, Gao W, Geng C, Zhou H, Leng Y, Li Y, Chen W (2017) Arsenic trioxide potentiates sensitivity of multiple myeloma cells to lenalidomide by upregulating cereblon expression levels. Oncol Lett 14:3243–3248.  https://doi.org/10.3892/ol.2017.6502 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Department of BiochemistryAll India Institute of Medical Sciences (AIIMS)New DelhiIndia
  2. 2.Department of BiochemistryNDMC Medical College & Hindu Rao HospitalNew DelhiIndia

Personalised recommendations