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Acid Water-ground Nano-realgar Is Superior to Crude Realgar in Promoting Apoptosis of MCF-7 Breast Cancer Cells

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Abstract

Objective

Realgar is a traditional mineral Chinese medicine with antitumor effects, but it has high toxicity and low efficacy in its crude form. The purpose of this study was to optimize realgar to increase its efficacy and therapeutic potential.

Methods

Crude realgar (CR) was mechanically ground to obtain nano-realgar (NR), and then nano-realgar processed products (NRPPs) were obtained using three different traditional Chinese medicine processing methods: grinding in water, acid water, and alkali water, respectively.

Results

By analyzing the size distribution of nanoparticles and the content of arsenic trioxide (As2O3; ATO), we found that acid water-ground NRPPs had the characteristics of high purity and low toxicity. The effects of CR, NR, and NRPPs on proliferation, cell cycle, and apoptosis of MCF-7 cells were detected, and the ability of NRPPs to induce apoptosis in MCF-7 cells was analyzed. The results showed that the average particle size of acid water-ground NRPPs was 137.7 nm, and the content of ATO was 2.83 mg/g. Acid water-ground NRPPs showed better effects on inhibiting proliferation, cell cycle, and apoptosis of MCF-7 cells than CR and NR. Western blot assays further confirmed that acid water-ground NRPPs upregulated the protein expression of TP53, Bax, cytochrome c, caspase-9, and caspase-3 in MCF-7 cells (P<0.05) and inhibited the expression of Bcl-2 (P<0.05).

Conclusion

These results suggest that acid water-ground NRPPs can induce apoptosis of MCF-7 cells through regulating mitochondrial-mediated apoptosis, providing evidence for the clinical application of realgar.

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References

  1. Sung H, Ferlay J, Siegel RL, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin, 2021,71(3):209–249

    PubMed  Google Scholar 

  2. Jemal A, Siegel R, Xu J, et al. Cancer statistics, 2010. CA Cancer J Clin, 2010,60(5):277–300

    Article  PubMed  Google Scholar 

  3. Niu QY, Chen WP, Tian YZ, et al. Preparation of medicinal realgar using realgar tailings. Chem Environ Protect (Chinese), 2002,22(3):169–172

    CAS  Google Scholar 

  4. Yao CM, Xiong XX, Liu CT. Metallogenic Fluid Evolution and Mineralization of Shimen Realgar Deposit, Hunan. Deposit Geol (Chinese), 1998,17(26): 1063–1066

    Google Scholar 

  5. Niu M, Shen Y, Qi J, et al. Effects of realgar (As4S4) on degradation of PML-RARA harboring acquired arsenic-resistance mutations. Ann Hematol, 2017,96:1945–1948

    Article  CAS  PubMed  Google Scholar 

  6. Wang L, Zhou GB, Liu P, et al. Dissection of mechanisms of Chinese medicinal formula Realgar-Indigo naturalis as an effective treatment for promyelocytic leukemia. Proc Natl Acad Sci USA, 2008,105:4826–4831

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Liu Y, He P, Cheng X, et al. Long-term outcome of 31 cases of refractory acute promyelocytic leukemia treated with compound realgar natural indigo tablets administered alternately with chemotherapy. Oncol Lett, 2015,10:1184–1190

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Mao JH, Sun XY, Liu JX, et al. As4S4 targets RING-type E3 ligase c-CBL to induce degradation of BCR-ABL in chronic myelogenous leukemia. Proc Natl Acad Sci USA, 2010,107:21683–21688

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Yin T, Wu YL, Sun HP, et al. Combined effects of As4 S4 and imatinib on chronic myeloid leukemia cells and BCR-ABL oncoprotein. Blood, 2004,104:4219–4225

    Article  CAS  PubMed  Google Scholar 

  10. Wu JZ, Ho PC. Evaluation of the in vitro activity and in vivo bioavailability of realgar nanoparticles prepared by cryo-grinding. Eur J Pharm Sci, 2006,29:35–44

    Article  PubMed  CAS  Google Scholar 

  11. Liao Q, Ji L, Deng F, et al. Study on the variation rule of As2S2 and As2O3 content in different realgar processed products. Tradition Chin Med Clin (Chinese), 2013,4(2):21–24

    Google Scholar 

  12. Guo T, Shi F, Yang G, et al. Effect of solid dispersion technology on stability and in vitro dissolution of realgar nanoparticles. Chin J Tradit Chin Med (Chinese), 2013,38(17):2782–2787

    CAS  Google Scholar 

  13. Tian Y, Wang X, Xi R, et al. Enhanced antitumour activity of realgar mediated by milling it to nano-size. Int J Nanomed, 2014,9:745–757

    Google Scholar 

  14. Wang XB, Gao HY, Hou BL, et al. Nanoparticle realgar powders induce apoptosis in U937 cells through caspase MAPK and mitochondrial pathways. Arch Pharm Res, 2007,30(5):653–658

    Article  CAS  PubMed  Google Scholar 

  15. Wang YS, Zhou ST, Wei HL. Apoptosis effects of drug sensitivity leukemia cells induced by nano-realgar. China J Chin Materia Medica (Chinese), 2013,38(13):2202–2205

    Google Scholar 

  16. Wang XB, Xian RG, Shi Y, et al. Optimization study of nano-realgar preparation process. J Chin Pharm Sci of PLA (Chinese), 2008,24(06):471–474

    CAS  Google Scholar 

  17. Shi F, Feng NP, Omari-Siaw E. Realgar nanoparticle-based microcapsules: preparation and in-vitro/in-vitro characterizations. J Pharm Pharmacol, 2015,67(1):35–42

    Article  CAS  PubMed  Google Scholar 

  18. Zhao Y, Yuan B, Onda K, et al. Anticancer efficacies of arsenic disulfide through apoptosis induction, cell cycle arrest, and pro-survival signal inhibition in human breast cancer cells. Am J Cancer Res, 2018,8(3):366–386

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Zhao Y, Onda K, Sugiyama K, et al. Antitumour effects of arsenic disulfide on the viability, migratory ability, apoptosis and autophagy of breast cancer cells. Oncol Rep, 2019,41(1):27–42

    CAS  PubMed  Google Scholar 

  20. Khairul I, Wang QQ, Jiang YH, et al. Metabolism, toxicity and anticancer activities of arsenic compounds. Oncotarget, 2017,8(14):23905–23926

    Article  PubMed  PubMed Central  Google Scholar 

  21. Wu J, Shao Y, Liu J, et al. The medicinal use of realgar (As4S4) and its recent development as an anticancer agent. J Ethnopharmacol, 2011,135(3):595–602

    Article  CAS  PubMed  Google Scholar 

  22. Liu X, Li X, Wang L, et al. Realgar induces apoptosis in the chronic lymphocytic leukemia cell line MEC-1. Mol Med Rep, 2013,8(6):1866–1870

    Article  CAS  PubMed  Google Scholar 

  23. Yang MH, Wan WQ, Luo JS, et al. Multicenter randomized trial of arsenic trioxide and Realgar-Indigo naturalis formula in pediatric patients with acute promyelocytic leukemia: Interim results of the SCCLG-APL clinical study. Am J Hematol, 2018,93(12):1467–1473

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Zhao QH, Zhang Y, Liu Y, et al. Anticancer effect of realgar nanoparticles on mouse melanoma skin cancer in vivo via transdermal drug delivery. Med Oncol, 2010,27(2):203–212

    Article  CAS  PubMed  Google Scholar 

  25. Qi YF, Li HJ, Liu ZD. The effect of nano-realgar on the proliferation of lung cancer A549 cells and its mechanism. Chin J Cancer Prevent Treat (Chinese), 2013,20(01):27–30

    CAS  Google Scholar 

  26. Song P, Chen P, Wang D, et al. Realgar transforming solution displays anticancer potential against human hepatocellular carcinoma HepG2 cells by inducing ROS. Int J Oncol, 2017,50(2):660–670

    Article  CAS  PubMed  Google Scholar 

  27. Zhang MH, Chen JQ, Guo HM, et al. Combination of LC/MS and GC/MS based metabolomics to study the hepatotoxic effect of realgar nanoparticles in rats. Chin J Nat Med, 2017,15(9):684–694

    CAS  PubMed  Google Scholar 

  28. Wang G, Zhang T, Sun W, et al. Arsenic sulfide induces apoptosis and autophagy through the activation of ROS/JNK and suppression of Akt/mTOR signaling pathways in osteosarcoma. Free Radic Biol Med, 2017,106:24–37

    Article  CAS  PubMed  Google Scholar 

  29. Xi XX, Sun J, Xi DB, et al. Realgar Nanoparticles Inhibit Migration, Invasion and Metastasis in a Mouse Model of Breast Cancer by Suppressing Matrix Metalloproteinases and Angiogenesis. Curr Drug Deliv, 2020,17(2):148–158

    Article  CAS  Google Scholar 

  30. An YL, Nie F, Wang ZY, et al. Preparation and characterization of realgar nanoparticles and their inhibitory effect on rat glioma cells. Int J Nanomed, 2011,6:3187–3194

    Article  CAS  Google Scholar 

  31. Qin YU, Wang H, Liu ZY, et al. Realgar quantum dots induce apoptosis and necrosis in HepG2 cells through endoplasmic reticulum stress. Biomed Rep, 2015,3(5):657–662

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Ding W, Zhang L, Kim S, et al. Arsenic sulfide as a potential anti-cancer drug. Mol Med Rep, 2015,11:968–974

    Article  CAS  PubMed  Google Scholar 

  33. Tse WP, Cheng CH, Che CT, et al. Arsenic trioxide, arsenic pentoxide, and arsenic iodide inhibit human keratinocyte proliferation through the induction of apoptosis. J Pharmacol Exp Ther, 2008,326(2):388–94

    Article  CAS  PubMed  Google Scholar 

  34. Zhao WZ, Lu X, Yuan Y, et al. Effect of size and processing method on the cytotoxicity of realgar nanoparticles in cancer cell lines. Int J Nanomed, 2011,6:1569–1577

    Article  CAS  Google Scholar 

  35. Wang Y, Zhang Y, Yang L, et al. Arsenic trioxide induces the apoptosis of human breast cancer MCF-7 cells through activation of caspase-3 and inhibition of HERG channels. Exp Ther Med, 2011,2(3):481–486

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  36. Shi Y, Cao T, Huang H, Lian CQ, et al. Arsenic trioxide inhibits cell growth and motility via up-regulation of let-7a in breast cancer cells. Cell Cycle, 2017,16(24): 2396–2403

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Cheng YX, Liu R, Wang Q, et al. Realgar-induced apoptosis of cervical cancer cell line Siha via cytochrome c release and caspase-3 and caspase-9 activation. Chin J Inter Med, 2012,18:359–365

    Article  CAS  Google Scholar 

  38. Wang XB, Gao HY, Hou BL, et al. Nanoparticle realgar powders induce apoptosis in U937 cells through caspase MAPK and mitochondrial pathways. Arch Pharm Res, 2007,30:653–658

    Article  CAS  PubMed  Google Scholar 

  39. Liu R, Pu D, Liu Y, et al. Induction of SiHa cells apoptosis by nanometer realgar suspension and its mechanism. J Huazhong Univ Sci Technolog Med Sci, 2008,28(3):317–321

    Article  PubMed  CAS  Google Scholar 

  40. Wang S, He MF, Li LM, et al. Cell-in-Cell Death Is Not Restricted by Caspase-3 Deficiency in MCF-7 Cells. J Breast Cancer, 2016,19(3):231–241

    Article  PubMed  PubMed Central  Google Scholar 

  41. Changizi V, Azariasl S, Motevaseli E, et al. Assessment Synergistic Effects of Integrated Therapy with Epigallocatechin-3-Gallate (EGCG) & Arsenic Trioxide and Irradiation on Breast Cancer Cell Line. Iran J Public Health, 2020,49(8):1555–1563

    PubMed  PubMed Central  Google Scholar 

  42. Gao H, Zhang Y, Dong L, et al. Triptolide induces autophagy and apoptosis through ERK activation in human breast cancer MCF-7 cells. Exp Ther Med (Chinese), 2018,15(4):3413–3419

    CAS  Google Scholar 

Download references

Acknowledgements

We are grateful for the technical support provided by the Flow Cytometry Core of Hubei University of Chinese Medicine.

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Correspondence to Yu-xue Wang or Xiu-qiao Zhang.

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The authors declare that they have no competing interests.

Additional information

This study was generously supported by the Science and Technology Research Project of Hubei Education Department (No. B2019097).

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Xi, J., Fang, Jh., Xiong, Xm. et al. Acid Water-ground Nano-realgar Is Superior to Crude Realgar in Promoting Apoptosis of MCF-7 Breast Cancer Cells. CURR MED SCI 42, 720–732 (2022). https://doi.org/10.1007/s11596-022-2605-5

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  • DOI: https://doi.org/10.1007/s11596-022-2605-5

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