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Parasporin-4, a novel apoptosis inducer of breast cancer cells produced by Bacillus thuringiensis

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Abstract

Background

Parasporin (PS) proteins have cytocidal activity preferential for various human malignant cells. The purpose of this investigation was to see if the PS separated from B. thuringiensis strain E8 isolate had any particular cytotoxicity against breast cancer.

Methods and results

The extracted spores-crystal proteins were solubilized and digested with proteinase K. Cytotoxicity effects were analysed by MTT assay. Caspases activities were measured using ELISA. SDS-PAGE analysis was performed for determination of molecular weight of Cry protein. Identification of extracted proteins function was evaluated by MALDI-TOF MS analysis. Breast cancer cells line (MCF-7) was highly susceptible to 1 mg/mL PS and showed apoptosis characteristics, but it has no effects on the normal cells (HEK293). Apoptosis evaluation showed that caspases 1, 3, 9 and BAX were remarkably up-regulated in cancer cells, indicating the intrinsic pathway activation in these cells. PS Size was determined using SDS-PAGE in E8 isolate as 34 kDa and a 25 kDa digested peptide was identified as PS4. The function of PS4 was reported as an ABC-transporter by spectrometry.

Conclusion

The data of the present study show that PS4 is a selective cytotoxic protein against breast cancer and a molecule with a lot of potentials for next researches.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Katayama H, Kusaka Y, Yokota H, Akao T, Kojima M, Nakamura O, Mekada E, Mizuki E (2007) Parasporin-1, a novel cytotoxic protein from Bacillus thuringiensis, induces Ca2 + influx and a sustained elevation of the cytoplasmic Ca2 + concentration in toxin-sensitive cells. J Biol Chem 282(10):7742–7752. https://doi.org/10.1074/jbc.M611382200

    Article  CAS  PubMed  Google Scholar 

  2. Goleij Z, Mahmoodzadeh Hosseini H, Sedighian H, Behzadi E, Halabian R, Sorouri R, Imani Fooladi AA (2020) Breast cancer targeted/ therapeutic with double and triple fusion immunotoxins. J Steroid Biochem Mol Biol 200:105651. https://doi.org/10.1016/j.jsbmb.2020.105651

    Article  CAS  PubMed  Google Scholar 

  3. Moazamian E, Bahador N, Azarpira N, Rasouli M (2018) Anti-cancer parasporin toxins of new Bacillus thuringiensis against human Colon (HCT-116) and blood (CCRF-CEM) Cancer Cell Lines. Curr Microbiol 75(8):1090–1098. https://doi.org/10.1007/s00284-018-1479-z

    Article  CAS  PubMed  Google Scholar 

  4. García-Montelongo M, González-Villarreal SE, Del Rincón-Castro MC, Ibarra JE (2021) Use of RNAi as a preliminary tool for screening putative receptors of nematicidal toxins from Bacillus thuringiensis. Arch Microbiol 203(4):1649–1656. https://doi.org/10.1007/s00203-020-02179-1

    Article  CAS  PubMed  Google Scholar 

  5. Santos EN, Menezes LP, Dolabella SS, Santini A, Severino P, Capasso R, Zielinska A, Souto EB, Jain S (2022) Bacillus thuringiensis: from biopesticides to anticancer agents. Biochimie 192:83–90. https://doi.org/10.1016/j.biochi.2021.10.003

    Article  CAS  PubMed  Google Scholar 

  6. Rampersad J, Ammons D (2005) A Bacillus thuringiensis isolation method utilizing a novel stain, low selection and high throughput produced atypical results. BMC Microbiol 5(1):52. https://doi.org/10.1186/1471-2180-5-52

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Nadarajah VD, Ting D, Chan KK, Mohamed SM, Kanakeswary K, Lee HL (2008) Selective cytotoxic activity against leukemic cell lines from mosquitocidal Bacillus thuringiensis parasporal inclusions. Southeast Asian J Trop Med Public Health 39(2):235–245

    CAS  PubMed  Google Scholar 

  8. Mendoza-Almanza G, Esparza-Ibarra EL, Ayala-Luján JL, Mercado-Reyes M, Godina-González S, Hernández-Barrales M, Olmos-Soto J (2020) The Cytocidal Spectrum of Bacillus thuringiensis toxins: from insects to Human Cancer cells. Toxins 12(5). https://doi.org/10.3390/toxins12050301

  9. Brasseur K, Auger P, Asselin E, Parent S, Côté J-C, Sirois M (2015) Parasporin-2 from a new Bacillus thuringiensis 4R2 strain induces caspases activation and apoptosis in Human Cancer cells. PLoS ONE 10(8):e0135106. https://doi.org/10.1371/journal.pone.0135106

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Kamalabadi-Farahani M, Karimi R, Atashi A (2023) High percentage of Cancer Stem cells in metastatic locations: Upregulation of cicBIRC6 in highly metastatic breast Cancer Subline. Molecular Biology Reports 50: 1303–1309 (2023). https://doi.org/10.1007/s11033-022-08024-6

  11. Ghoncheh M, Pournamdar Z, Salehiniya H (2016) Incidence and mortality and epidemiology of breast Cancer in the World. Asian Pac J cancer prevention: APJCP 17(S3):43–46. https://doi.org/10.7314/apjcp.2016.17.s3.43

    Article  Google Scholar 

  12. Motaghed M, Al-Hassan FM, Hamid SS (2013) Cellular responses with thymoquinone treatment in human breast cancer cell line MCF-7. Pharmacognosy Res 5(3):200–206. https://doi.org/10.4103/0974-8490.112428

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Meissner K, Talsky N, Olliges E, Jacob C, Stötzer OJ, Salat C, Braun M, Flondor R (2019) Individual factors contributing to nausea in First-Time Chemotherapy Patients: a prospective cohort study. Front Pharmacol 10:410–410. https://doi.org/10.3389/fphar.2019.00410

    Article  PubMed  PubMed Central  Google Scholar 

  14. Cassileth BR, Deng G (2004) Complementary and alternative therapies for cancer. Oncologist 9(1):80–89. https://doi.org/10.1634/theoncologist.9-1-80

    Article  PubMed  Google Scholar 

  15. West H (2018) Complementary and alternative medicine in Cancer Care. JAMA Oncol 4(1):139–139. https://doi.org/10.1001/jamaoncol.2017.3120

    Article  PubMed  Google Scholar 

  16. Foster FM, Owens TW, Tanianis-Hughes J, Clarke RB, Brennan K, Bundred NJ, Streuli CH (2009) Targeting inhibitor of apoptosis proteins in combination with ErbB antagonists in breast cancer. Breast Cancer Res 11(3):R41. https://doi.org/10.1186/bcr2328

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ammons D, Rampersad J, Khan A (2002) Usefulness of staining parasporal bodies when screening for Bacillus thuringiensis. J Invertebr Pathol 79(3):203–204. https://doi.org/10.1016/s0022-2011(02)00018-6

    Article  PubMed  Google Scholar 

  18. Yasutake K, Binh ND, Kagoshima K, Uemori A, Ohgushi A, Maeda M, Mizuki E, Yu YM, Ohba M (2006) Occurrence of parasporin-producing Bacillus thuringiensis in Vietnam. Can J Microbiol 52(4):365–372. https://doi.org/10.1139/w05-134

    Article  CAS  PubMed  Google Scholar 

  19. Laemmli UK (1970) Cleavage of structural proteins during the Assembly of the Head of Bacteriophage T4. Nature 227(5259):680–685. https://doi.org/10.1038/227680a0

    Article  CAS  PubMed  Google Scholar 

  20. Bahmani S, Azarpira N, Moazamian E (2019) Anti-colon cancer activity of Bifidobacterium metabolites on colon cancer cell line SW742. Turkish J Gastroenterol 30(9):835–842. https://doi.org/10.5152/tjg.2019.18451

    Article  Google Scholar 

  21. Bringans S, Eriksen S, Kendrick T, Gopalakrishnakone P, Livk A, Lock R, Lipscombe R (2008) Proteomic analysis of the venom of Heterometrus longimanus (asian black scorpion). Proteomics 8(5):1081–1096. https://doi.org/10.1002/pmic.200700948

    Article  CAS  PubMed  Google Scholar 

  22. Khorramnejad A, Gomis-Cebolla J, Talaei-Hassanlouei R, Bel Y, Escriche B (2020) Genomics and Proteomics analyses revealed novel candidate Pesticidal Proteins in a lepidopteran-toxic Bacillus thuringiensis strain. Toxins 12(11):673

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. Cancer J Clin 68(6):394–424. https://doi.org/10.3322/caac.21492

    Article  Google Scholar 

  24. Nafissi N, Khayamzadeh M, Zeinali Z, Pazooki D, Hosseini M, Akbari ME (2018) Epidemiology and histopathology of breast Cancer in Iran versus other Middle Eastern Countries. Middle East Journal of Cancer 9(3):243–251. https://doi.org/10.30476/mejc.2018.42130

    Article  Google Scholar 

  25. Hermelink K (2015) Chemotherapy and cognitive function in breast Cancer patients: the So-Called chemo brain. J Natl Cancer Inst Monogr 2015(51):67–69. https://doi.org/10.1093/jncimonographs/lgv009

    Article  PubMed  Google Scholar 

  26. Okumura S, Saitoh H, Ishikawa T, Wasano N, Yamashita S, Kusumoto K-i, Akao T, Mizuki E, Ohba M, Inouye K (2005) Identification of a Novel cytotoxic protein, Cry45Aa, from Bacillus thuringiensis A1470 and its selective cytotoxic activity against various mammalian cell lines. J Agric Food Chem 53(16):6313–6318. https://doi.org/10.1021/jf0506129

    Article  CAS  PubMed  Google Scholar 

  27. Amano H, Yamagiwa M, Akao T, Mizuki E, Ohba M, Sakai H (2005) A novel 29-kDa crystal protein from Bacillus thuringiensis induces caspase activation and cell death of Jurkat T cells. Bioscience, Biotechnology, and Biochemistry. 69:2063–2072. https://doi.org/10.1271/bbb.69.2063. 11

  28. Beena V, Ramnath V, Sreekumar KP, Karthiayini K, Philomina PT, Girija D (2019) Crystal protein of a novel Bacillus thuringiensis strain inducing cell cycle arrest and apoptotic cell death in human leukemic cells. Sci Rep 9(1):9661. https://doi.org/10.1038/s41598-019-45928-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Ekino K, Okumura S, Ishikawa T, Kitada S, Saitoh H, Akao T, Oka T, Nomura Y, Ohba M, Shin T, Mizuki E (2014) Cloning and characterization of a unique cytotoxic protein parasporin-5 produced by Bacillus thuringiensis A1100 strain. Toxins 6(6):1882–1895. https://doi.org/10.3390/toxins6061882

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Chubicka T, Girija D, Deepa K, Salini S, Meera N, Raghavamenon AC, Divya MK, Babu TD (2018) A parasporin from Bacillus thuringiensis native to Peninsular India induces apoptosis in cancer cells through intrinsic pathway. J Biosci 43(2):407–416

    Article  CAS  PubMed  Google Scholar 

  31. Aboul-Soud MAM, Al-Amri MZ, Kumar A, Al-Sheikh YA, Ashour AE, El-Kersh TA (2019) Specific cytotoxic Effects of Parasporal Crystal Proteins isolated from native saudi arabian Bacillus thuringiensis strains against Cervical Cancer cells. Molecules 24(3):506. https://doi.org/10.3390/molecules24030506

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Kushwaha PP, Singh AK, Prajapati KS, Shuaib M, Fayez S, Bringmann G, Kumar S (2020) Induction of apoptosis in breast cancer cells by naphthylisoquinoline alkaloids. Toxicol Appl Pharmcol 409:115297. https://doi.org/10.1016/j.taap.2020.115297

    Article  CAS  Google Scholar 

  33. Rubio-Infante N, Ilhuicatzi-Alvarado D, Torres-Martínez M, Reyes-Grajeda JP, Nava-Acosta R, González-González E, Moreno-Fierros L (2018) The Macrophage Activation Induced by Bacillus thuringiensis Cry1Ac Protoxin involves ERK1/2 and p38 pathways and the Interaction with cell-Surface-HSP70. J Cell Biochem 119(1):580–598. https://doi.org/10.1002/jcb.26216

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Microbiology, College of Sciences, Agriculture and Modern Technology, Shiraz Branch, Islamic Azad University, Shiraz, Iran

    Roya Rezaei & Elham Moazamian

  2. Shiraz Endocrine and Metabolism Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

    Nima Montazeri-Najafabady

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Contributions

Elham Moazamian participated in the design of the study. Elham Moazamian and Roya Rezaei performed experiments, analysed data and drafted the manuscript. Nima Montazeri-Najafabady contributed to writing, drafting and submitting the manuscript. All authors reviewed and contributed to final editing and refinement of the manuscript.

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Correspondence to Elham Moazamian.

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Rezaei, R., Moazamian, E. & Montazeri-Najafabady, N. Parasporin-4, a novel apoptosis inducer of breast cancer cells produced by Bacillus thuringiensis. Mol Biol Rep 50, 4469–4480 (2023). https://doi.org/10.1007/s11033-023-08378-5

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