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Synergistic effects of tanshinone IIA and andrographolide on the apoptosis of cancer cells via crosstalk between p53 and reactive oxygen species pathways

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Abstract

Background

Tanshinone IIA (Tan IIA) and andrographolide (Andro) are natural compounds that are reported to exhibit anticancer activities against various types of cancers. The aim of this study is to evaluate the synergistic anticancer effects of the combination of Tan IIA and Andro, and to investigate the mechanisms of pharmacological effect and their potential applications as an anticancer therapy in clinics.

Methods

The anticancer effects of the combination of Tan IIA and Andro on MCF7, SMMC7721, and BGC823 cells were explored. The apoptosis of the cancer cells was determined by MTT and AV-PI dual stain assays. The intracellular GSH level was measured by DTNB assay, and the intracellular levels of reactive oxygen species (ROS) were examined by flow cytometry. The expression of the proteins in the apoptosis pathway was determined by immunobloting.

Results

The combination of Tan IIA and Andro exhibited significant synergistic anticancer effects against cancer cells, especially at low concentrations. Andro reacted with the thiol group of intracellular GSH, thus disrupting the GSH redox cycle and eventually increasing the level of intracellular ROS. Tan IIA triggered p53 responses and apoptosis by binding to the DNA of cancer cells. The crosstalk between ROS and p53 exhibited a synergistic effect on the apoptosis of cancer cells.

Conclusion

The combination of Tan IIA and Andro showed significant synergistic effects on cancer cell apoptosis by promoting crosstalk between ROS and p53, providing a novel and effective combination that has the potential to be applied in clinical anticancer therapy.

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Abbreviations

Andro:

Andrographolide

AV-PI:

Annexin V-fluorescein isothiocyanate-propidium iodide

BSO:

Buthionine sulfoximine

CI:

Combination index

DETC:

Diethylthiocarbamate

DCF:

Dichlorofluorescein

DHE:

Dihydroethidium

DMSO:

Dimethyl sulfoxide

DTNB:

5,5′-dithiobis(2-nitrobenzoic acid)

GSH:

Glutathione

HPLC:

High performance liquid chromatography

MTT:

3-(4,5-dimethylthiazol-2 -yl)-2,5-diphenyl tetrazolium bromide

NAC:

N-acetyl-l-cysteine

PFT-α:

1-(4-methylphenyl)-2-(4,5,6,7-tetrahydro-2-imino-3(2H)-benzothiazolyl) ethanone hydrobromide

PVDF:

Polyvinylidene fluoride

ROS:

Reactive oxygen species

Tan IIA:

Tanshinone IIA

References

  1. Xu Y, Xin Y, Diao Y, Lu C, Fu J, Luo L, et al. Synergistic effects of apigenin and paclitaxel on apoptosis of cancer cells. PLoS ONE. 2011;6(12):e29169.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Tian XY, Liu L. Drug discovery enters a new era with multi-target intervention strategy. Chin J Integr Med. 2012;18(7):539–42.

    PubMed  Google Scholar 

  3. Ulrich-Merzenich GS. Combination screening of synthetic drugs and plant derived natural products—potential and challenges for drug development. Synergy. 2014;1(1):59–69.

    Google Scholar 

  4. Gao S, Liu Z, Li H, Little PJ, Liu P, Xu S. Cardiovascular actions and therapeutic potential of tanshinone IIA. Atherosclerosis. 2012;220(1):3–10.

    CAS  PubMed  Google Scholar 

  5. Xu S, Liu P. Tanshinone II-A: new perspectives for old remedies. Expert Opin Ther Pat. 2013;23(2):149–53.

    CAS  PubMed  Google Scholar 

  6. Sung HJ, Choi SM, Yoon Y, An KS. Tanshinone IIA, an ingredient of Salvia miltiorrhiza BUNGE, induces apoptosis in human leukemia cell lines through the activation of caspase-3. Exp Mol Med. 1999;31(4):174–8.

    CAS  PubMed  Google Scholar 

  7. Liu JJ, Lin DJ, Liu PQ, Huang M, Li XD, Huang RW. Induction of apoptosis and inhibition of cell adhesive and invasive effects by tanshinone IIA in acute promyelocytic leukemia cells in vitro. J Biomed Sci. 2006;13(6):813–23.

    CAS  PubMed  Google Scholar 

  8. Lin C, Wang L, Wang H, Yang L, Guo H, Wang X. Tanshinone IIA inhibits breast cancer stem cells growth in vitro and in vivo through attenuation of IL-6/STAT3/NF-kB signaling pathways. J Cell Biochem. 2013;114(9):2061–70.

    CAS  PubMed  Google Scholar 

  9. Yu T, Zhou Z, Mu Y, de Lima LG, Luo KQ. A novel anti-cancer agent, acetyltanshinone IIA, inhibits oestrogen receptor positive breast cancer cell growth by down-regulating the oestrogen receptor. Cancer Lett. 2014;346(1):94–103.

    CAS  PubMed  Google Scholar 

  10. Chiu SC, Huang SY, Chen SP, Su CC, Chiu TL, Pang CY. Tanshinone IIA inhibits human prostate cancer cells growth by induction of endoplasmic reticulum stress in vitro and in vivo. Prostate Cancer Prostatic Dis. 2013;16(4):315–22.

    CAS  PubMed  Google Scholar 

  11. Su CC, Chen GW, Kang JC, Chan MH. Growth inhibition and apoptosis induction by tanshinone IIA in human colon adenocarcinoma cells. Planta Med. 2008;74(11):1357–62.

    CAS  PubMed  Google Scholar 

  12. Yuan SL, Wei YQ, Wang XJ, Xiao F, Li SF, Zhang J. Growth inhibition and apoptosis induction of tanshinone II-A on human hepatocellular carcinoma cells. World J Gastroenterol. 2004;10(14):2024–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Li Q, Wang Y, Feng N, Fan Z, Sun J, Nan Y. Novel polymeric nanoparticles containing tanshinone IIA for the treatment of hepatoma. J Drug Target. 2008;16(10):725–32.

    PubMed  Google Scholar 

  14. Lai YH, Yu SL, Chen HY, Wang CC, Chen HW, Chen JJ. The HLJ1-targeting drug screening identified Chinese herb andrographolide that can suppress tumour growth and invasion in non-small-cell lung cancer. Carcinogenesis. 2013;34(5):1069–80.

    CAS  PubMed  Google Scholar 

  15. Xie J, Liu J, Liu H, Liang S, Lin M, Gu Y, et al. The antitumor effect of tanshinone IIA on anti-proliferation and decreasing VEGF/VEGFR2 expression on the human non-small cell lung cancer A549 cell line. Acta Pharm Sin B. 2015;5(6):554–63.

    PubMed  PubMed Central  Google Scholar 

  16. Cheng CY, Su CC. Tanshinone IIA may inhibit the growth of small cell lung cancer H146 cells by up-regulating the Bax/Bcl-2 ratio and decreasing mitochondrial membrane potential. Mol Med Rep. 2010;3(4):645–50.

    CAS  PubMed  Google Scholar 

  17. Jang, S.I., Kim, H.J., Kim, Y.J., Jeong, S.I., You, Y.O. Tanshinone IIA inhibits LPS-induced NF-kappaB activation in RAW 264.7 cells: possible involvement of the NIK-IKK, ERK1/2, p38 and JNK pathways. EUR J PHARMACOL. 2006;542(1-3):1-7.

  18. Wang X, Wei Y, Yuan S, Liu G, Lu Y, Zhang J, et al. Potential anticancer activity of tanshinone IIA against human breast cancer. Int J Cancer. 2005;116(5):799–807.

    CAS  PubMed  Google Scholar 

  19. Zhang Z, Gao J, Wang Y, Song T, Zhang J, Wu G, et al. Tanshinone IIA triggers p53 responses and apoptosis by RNA polymerase II upon DNA minor groove binding. Biochem Pharmacol. 2009;78(10):1316–22.

    CAS  PubMed  Google Scholar 

  20. Li J, Cheung HY, Zhang Z, Chan GK, Fong WF. Andrographolide induces cell cycle arrest at G2/M phase and cell death in HepG2 cells via alteration of reactive oxygen species. Eur J Pharmacol. 2007;568(1–3):31–44.

    CAS  PubMed  Google Scholar 

  21. Calabrese C, Berman SH, Babish JG, Ma X, Shinto L, Dorr M, et al. A phase I trial of andrographolide in HIV positive patients and normal volunteers. Phytother Res. 2000;14(5):333–8.

    CAS  PubMed  Google Scholar 

  22. Jayakumar T, Hsieh CY, Lee JJ, Sheu JR. Experimental and clinical pharmacology of andrographis paniculata and its major bioactive phytoconstituent andrographolide. Evid Based Complement Alternat Med. 2013;2013:846740.

    PubMed  PubMed Central  Google Scholar 

  23. Chen W, Feng L, Nie H, Zheng X. Andrographolide induces autophagic cell death in human liver cancer cells through cyclophilin D-mediated mitochondrial permeability transition pore. Carcinogenesis. 2012;33(11):2190–8.

    CAS  PubMed  Google Scholar 

  24. Chao CY, Lii CK, Hsu YT, Lu CY, Liu KL, Li CC, et al. Induction of heme oxygenase-1 and inhibition of TPA-induced matrix metalloproteinase-9 expression by andrographolide in MCF-7 human breast cancer cells. Carcinogenesis. 2013;34(8):1843–51.

    CAS  PubMed  Google Scholar 

  25. Bao GQ, Shen BY, Pan CP, Zhang YJ, Shi MM, Peng CH. Andrographolide causes apoptosis via inactivation of STAT3 and Akt and potentiates antitumor activity of gemcitabine in pancreatic cancer. Toxicol Lett. 2013;222(1):23–35.

    CAS  PubMed  Google Scholar 

  26. Jada SR, Matthews C, Saad MS, Hamzah AS, Lajis NH, Stevens MF, et al. Benzylidene derivatives of andrographolide inhibit growth of breast and colon cancer cells in vitro by inducing G(1) arrest and apoptosis. Br J Pharmacol. 2008;155(5):641–54.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Zhou J, Zhang S, Ong CN, Shen HM. Critical role of pro-apoptotic Bcl-2 family members in andrographolide-induced apoptosis in human cancer cells. Biochem Pharmacol. 2006;72(2):132–44.

    CAS  PubMed  Google Scholar 

  28. Sheeja K, Kuttan G. Activation of cytotoxic T lymphocyte responses and attenuation of tumor growth in vivo by andrographis paniculata extract and andrographolide. Immunopharmacol Immunotoxicol. 2007;29(1):81–93.

    CAS  PubMed  Google Scholar 

  29. Shi MD, Lin HH, Lee YC, Chao JK, Lin RA, Chen JH. Inhibition of cell-cycle progression in human colorectal carcinoma Lovo cells by andrographolide. Chem Biol Interact. 2008;174(3):201–10.

    CAS  PubMed  Google Scholar 

  30. Wang J, Tan XF, Nguyen VS, Yang P, Zhou J, Gao M, et al. A quantitative chemical proteomics approach to profile the specific cellular targets of andrographolide, a promising anticancer agent that suppresses tumor metastasis. Mol Cell Proteomics. 2014;13(3):876–86.

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Jones DP. Redefining oxidative stress. Antioxid Redox Signal. 2006;8(9–10):1865–79.

    CAS  Google Scholar 

  32. Martindale JL, Holbrook NJ. Cellular response to oxidative stress: signaling for suicide and survival. J Cell Physiol. 2002;192(1):1–15.

    CAS  PubMed  Google Scholar 

  33. Lim YP, Lim TT, Chan YL, Song AC, Yeo BH, Vojtesek B, et al. The p53 knowledgebase: an integrated information resource for p53 research. Oncogene. 2007;26(11):1517–21.

    CAS  PubMed  Google Scholar 

  34. Liu B, Chen Y, St CD. ROS and p53: a versatile partnership. Free Radic Biol Med. 2008;44(8):1529–35.

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Xie J, Zhang J, Wu H, Tang X, Liu J, Cheng G, et al. The influences of age on T lymphocyte subsets in C57BL/6 mice. Saudi J Biol Sci. 2017;24(1):108–13.

    CAS  PubMed  Google Scholar 

  36. Wu X, Zhu Y, Yan H, Liu B, Li Y, Zhou Q, et al. Isothiocyanates induce oxidative stress and suppress the metastasis potential of human non-small cell lung cancer cells. BMC Cancer. 2010;10(1):1–11.

    CAS  Google Scholar 

  37. Curtin JF, Donovan M, Cotter TG. Regulation and measurement of oxidative stress in apoptosis. J Immunol Methods. 2002;265(1–2):49–72.

    CAS  PubMed  Google Scholar 

  38. Zhao H, Kalivendi S, Zhang H, Joseph J, Nithipatikom K, Vasquez-Vivar J, et al. Superoxide reacts with hydroethidine but forms a fluorescent product that is distinctly different from ethidium: potential implications in intracellular fluorescence detection of superoxide. Free Radic Biol Med. 2003;34(11):1359–68.

    CAS  PubMed  Google Scholar 

  39. Chou TC. Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev. 2006;58(3):621–81.

    CAS  PubMed  Google Scholar 

  40. Ferret PJ, Soum E, Negre O, Fradelizi D. Auto-protective redox buffering systems in stimulated macrophages. BMC Immunol. 2002;3:3.

    PubMed  PubMed Central  Google Scholar 

  41. Ju W, Wang X, Shi H, Chen W, Belinsky SA, Lin Y. A critical role of luteolin-induced reactive oxygen species in blockage of tumor necrosis factor-activated nuclear factor-κB pathway and sensitization of apoptosis in lung cancer cells. Mol Pharmacol. 2007;71(5):1381.

    CAS  PubMed  Google Scholar 

  42. Misra UK, Pizzo SV. PFT-alpha inhibits antibody-induced activation of p53 and pro-apoptotic signaling in 1-LN prostate cancer cells. Biochem Biophys Res Commun. 2010;391(1):272–6.

    CAS  PubMed  Google Scholar 

  43. Shang Q, Xu H, Huang L. Tanshinone IIA: a promising natural cardioprotective agent. Evid Based Complement Alternat Med. 2012;2012:716459.

    PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors are grateful for the financial support of the National Natural Science Foundation of China (Grant No. 21376039), and Panjin Campus for Food Science and Technology Research Initiative, Dalian University of Technology.

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Authors and Affiliations

  1. School of Life Science and Biotechnology, Dalian University of Technology, NO. 2 Linggong Road, Ganjingzi District, Dalian, 116024, China

    Yingxue Li, Bo Jiang, Jingyun Wang & Yongming Bao

  2. School of Food and Environmental Science and Engineering, Dalian University of Technology, Panjin, 124221, China

    Yongming Bao

  3. Shennong Institute of Chinese Medicine, Guangzhou Union Biotech Co. Ltd, Guangzhou, 510663, China

    Yingxue Li & Ruiyin Wang

  4. Department of Occupational and Environmental Health, Dalian Medical University, Dalian, 116044, China

    Yachen Li

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  1. Yingxue Li
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Li, Y., Jiang, B., Wang, R. et al. Synergistic effects of tanshinone IIA and andrographolide on the apoptosis of cancer cells via crosstalk between p53 and reactive oxygen species pathways. Pharmacol. Rep 72, 400–417 (2020). https://doi.org/10.1007/s43440-019-00006-z

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