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Surface ultrastructure, protein profile and zymography of Blastocystis species isolated from patients with colorectal carcinoma

Journal of Parasitic Diseases volume 43, pages 294–303 (2019)Cite this article

Abstract

Oncogenic potential of Blastocystis species have been predicted on reporting the enhanced proliferation of human colorectal cancer cells by the parasite solubilized antigen in vitro, and the enhanced drug-induced carcinogenesis by infection in vivo. The present study is the first to investigate some phenotypic characters, namely the surface ultrastructure, protein profiles and protease activity of Blastocystis sp. isolated from three different clinical groups: colorectal carcinoma (CRC) patients, symptomatic and asymptomatic infected persons. Under SEM, all CRC Blastocystis sp. isolates had a very rough intensely folded surface in comparison to the less rough and completely smooth surface of all symptomatic and asymptomatic Blastocystis sp. Non-CRC isolates, respectively. Under reducing conditions, the sodium dodecyl sulfate–polyacrylamide gel electrophoresis had shown a significant presence of 2 protein bands of 230 and 32 KDa in 42.9% of Blastocystis sp. CRC isolates with their complete absence from Non-CRC isolates. While using non-reducing condition with the incorporation of gelatin in the gel to study the protease activity of the parasite, no significant difference existed between isolates of the three groups. In conclusion, the significant difference in surface ultrastructure and in protein profiles exists between Blastocystis sp. of CRC and Non-CRC isolates. These differences might be either secondary to the altered gut environment in the presence of CRC or are indicators of a different pathogenic potential of the parasite isolates inducing malignancy.

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References

  • Abaza SM, Rayan HZ, Soliman RH, Nemr NA, Mokhtar AB (2014) Subtype analysis of Blastocystis sp. isolates from symptomatic and asymptomatic patients in Suez Canal University Hospitals Ismailia Egypt. PUJ 7:56–67

    Google Scholar 

  • AbdelHameed DM, Hassanin OM (2011) Proteases activity of Blastocystis hominis subtype 3 in symptomatic and asymptomatic patients. Parasitol Res 109:321–327

    Article  Google Scholar 

  • AbouGamra MM, Elwakil HS, El Deeb HK, Khalifa KE, Abdel-Hafiz HE (2011) The potential use of 29 kDa protein as a marker of pathogenicity and diagnosis of symptomatic infections with Blastocystis hominis. Parasitol Res 108(5):1139–1146

    Article  Google Scholar 

  • Abu El-Fetouh NI, Abdelmegeed ES, Attia RA, El-Dosoky I, Azab MS (2015) Genotyping of Blastocystis hominis symptomatic isolates and kinetics of associated local CD3 and CD20 cell infiltrate. PUJ 8:115–122

    Google Scholar 

  • Alfellani MA, Stensvold CR, Vidal-Lapiedra A, Onuoha ES, Fagbenro-Beyioku AF, Clark CG (2013) Variable geographic distribution of Blastocystis subtypes and its potential implications. Acta Trop 126:11–18

    Article  PubMed  Google Scholar 

  • Arisue N, Hashimoto T, Yoshikawa H, Nakamura Y, Nakamura G, Nakamura F (2002) Phylogenetic position of Blastocystis hominis and of stramenopiles inferred from multiple molecular sequence data. J Eukaryot Microbiol 49:42–53

    Article  CAS  PubMed  Google Scholar 

  • Boreham PFL, Stenzel DJ (1993) Blastocystis in humans and animals: morphology, biology, and epizootiology. Adv Parasitol 32:1–70

    Article  CAS  PubMed  Google Scholar 

  • Bradford MM (1976) Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Cassidy MF, Stenzel DJ, Boreham PFL (1994) Electron microscopy of surface structures of Blastocystis sp. from different hosts. Parasitol Res 80:505–511

    Article  CAS  PubMed  Google Scholar 

  • Chan KH, Chandramathi S, Suresh K, Chua KH, Kuppusamy UR (2012) Effects of symptomatic and asymptomatic isolates of Blastocystis hominis on colorectal cancer cell line, HCT116. Parasitol Res 110:2475–2480

    Article  PubMed  Google Scholar 

  • Chandramathi S, Suresh K, Kuppusamy UR (2010) Solubilized antigen of Blastocystis hominis facilitates the growth of human colorectal cancer cells HCT116. Parasitol Res 106(4):941–945

    Article  PubMed  Google Scholar 

  • Chen X, Singh M, Ho L, Tan S, Yap E (1999) Characterization of protein profiles and cross-reactivity of Blastocystis antigens by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and immunoblotting analysis. Parasitol Res 85:343–346

    Article  CAS  PubMed  Google Scholar 

  • Clark CG, Diamond LS (2002) Methods for cultivation of luminal parasitic protists of clinical importance. Clin Microbiol Rev 15(3):329–341

    Article  PubMed  PubMed Central  Google Scholar 

  • Dunn LA, Boreham PFL, Stenzel DJ (1989) Ultra-structural variation of Blastocystis hominis stocks in culture. Int J Parasitol 19:43–56

    Article  CAS  PubMed  Google Scholar 

  • Eida AM, Eida MM (2008) Identification of Blastocystis hominis in patients with irritable bowel syndrome using microscopy and culture compared to PCR. PUJ 1(2):87–92

    Google Scholar 

  • El-Gayar EK, Mahmoud MM (2014) Do protozoa play a role in carcinogenesis? PUJ 7:80–85

    Google Scholar 

  • El-Wakil HS, Talaat RM (2009) Genetic analysis of Blastocystis hominis isolated from symptomatic and asymptomatic human hosts in Egypt. J Egypt Soc Parasitol 39:99–109

    Google Scholar 

  • Fadl HO, El-Akkad DMH, Abd El-Fattah DS, El-Bolaky HA, El-Bassiouni SO (2016) Study of the protein profiles of Blastocystis isolates from symptomatic and asymptomatic subjects. Med J Cairo Univ 84(3):349–353

    Google Scholar 

  • Garcia LS (2015) Diagnostic medical Parasitology, 6th edn. ASM Press, Washington

    Google Scholar 

  • Gericke A, Burchard G, Knobloch J, Walderich B (1997) Isoenzyme patterns of Blastocystis hominis patient isolates derived from symptomatic and healthy carriers. Trop Med Int Health 2(3):245–253

    Article  CAS  PubMed  Google Scholar 

  • Hegazy MM, Maklouf LM, El Hamshary EM, Dawoud HA, Eida AM (2008) Protein profile and morphometry of cultured human Blastocystis hominis from children with gastroenteritis and healthy ones. J Egypt Soc Parasitol 38(2):453–464

    CAS  PubMed  Google Scholar 

  • Ho LC, Armiugam A, Jeyaseelan K, Yap EH, Singh M (2000) Blastocystis elongation factor-1alpha: genomic organization, taxonomy, and phylogenetic relationships. Parasitology 121:135–144

    Article  CAS  PubMed  Google Scholar 

  • Init I, Mak J, Top S, Zulhainan Z, Prummon-Gkol S, Nissapatorn V, Wan-Yussof W, Anuar A (2003) Polypeptides associated with in vitro cyst formation of Blastocystis hominis. Southeast Asian J Trop Med Public Health 34:727–732

    CAS  PubMed  Google Scholar 

  • Kumarasamy V, Kuppusamy UR, Jayalakshmi P, Samudi C, Ragavan ND, Kumar S (2017) Exacerbation of colon carcinogenesis by Blastocystis sp. PLoS ONE 12(8):e0183097

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Laemmli UK (1970) Cleavage of structure proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Lanuza MD, Carbajal JA, Villar J, Mir A, Borrás R (1999) Soluble protein and antigenic heterogeneity in axenic Blastocystis hominis isolates: pathogenic implications. Parasitol Res 85:93–97

    Article  CAS  PubMed  Google Scholar 

  • Mohamed AM, Ahmed MA, Ahmed SA, Al-Semany SA, Alghamdi SS, Zaglool DA (2017) Predominance and association risk of Blastocystis hominis subtype I in colorectal cancer: a case-control study. Infect Agents Cancer 12:21

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Müller H (1994) Four serologically different groups within the species of Blastocystis hominis. Zentralbl Bakteriol 280:403–408

    Article  PubMed  Google Scholar 

  • Nagel R, Cuttell L, Stensvold CR, Mills PC, Bielefeldt-Ohmann H, Traub RJ (2012) Blastocystis subtypes in symptomatic and asymptomatic family members and pets and response to therapy. Intern Med J42:1187–1195

    Article  Google Scholar 

  • Parija SC, Jeremiah SS (2013) Symposium on Blastocystis: taxonomy, biology, and virulence. Trop Parasitol 3:17–25

    Article  PubMed  PubMed Central  Google Scholar 

  • Poirier P, Wawrzyniak I, Viverès CP, Delbac F, El Alaoui H (2012) New insights into Blastocystis sp.: a potential link with irritable bowel syndrome. PLoS Pathog 8(3):e1002545

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ragavan ND, Govind SK, Chye TT, Mahadeva S (2014) Phenotypic variation in Blastocystis sp. ST3. Parasites Vectors 7:404

    Article  PubMed  PubMed Central  Google Scholar 

  • Ramírez JD, Sánchez A, Hernández C, Flórez C, Bernal MC, Giraldo JC, Reyes P, López MC, García L, Cooper PJ et al (2016) Geographic distribution of human Blastocystis subtypes in South America. Infect Genet Evol 41:32–35

    Article  PubMed  Google Scholar 

  • Scanlan PD (2012) Blastocystis past pit falls and future perspectives. Trends Parasitol 28:327–334

    Article  PubMed  Google Scholar 

  • Silberman JD, Sogin ML, Leipe DD, Clark CG (1996) Human parasite finds taxonomic home. Nature 380:398

    Article  CAS  PubMed  Google Scholar 

  • Souppart L, Sanciu G, Cian A, Wawrzyniak I, Delbac F, Capron M et al (2009) Molecular epidemiology of human Blastocystis isolates in France. Parasitol Res 105:413–421

    Article  PubMed  Google Scholar 

  • Steer H (2007) Blastocystis hominis and Colorectal Cancer. Ann R Coll Surg Engl 89:539

    Article  PubMed Central  Google Scholar 

  • Stensvold CR (2013) Blastocystis-genetic diversity and molecular methods for diagnosis and epidemiology. Trop Parasitol 3:26–34

    Article  PubMed  PubMed Central  Google Scholar 

  • Stensvold CR, Arendrup MC, Jespersgaard C, Mølbak K, Nielsen HV (2007a) Detecting Blastocystis using parasitologic and DNA-based methods: a comparative study. Diagn Microbiol Infect Dis 59:303–307

    Article  CAS  PubMed  Google Scholar 

  • Stensvold CR, Suresh GK, Tan KS, Thompson RC, Traub RJ et al (2007b) Terminology for Blastocystis subtypes a consensus. Trends Parasitol 23:93–96

    Article  PubMed  Google Scholar 

  • Suresh K, Howe J, Chong SY, Ng GC, Ho LC, Loh AK, Ramachandran NP, Yap EH, Singh M (1994) Ultra-structural changes during in vitro encystment of Blastocystis hominis. Parasitol Res 80:327–335

    Article  CAS  PubMed  Google Scholar 

  • Tan KS (2008) New insights on classification, identification, and clinical relevance of Blastocystis sp. Clin Microbiol Rev 21:639–665

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tan TC, Suresh KG (2006) Predominance of amoeboid forms of Blastocystis hominis in isolates from symptomatic patients. Parasitol Res 98:189–193

    Article  CAS  PubMed  Google Scholar 

  • Tan TC, Suresh KG, Smith HV (2008) Phenotypic and genotypic characterization of Blastocystis hominis isolates implicates subtype 3 as a subtype with pathogenic potential. Parasitol Res 104:85–93

    Article  CAS  PubMed  Google Scholar 

  • Toth M, Fridman R (2001) Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography. Methods Mol Med 57(10):160–163

    Google Scholar 

  • Upcroft JA, Dunn LA, Dommet LS, Healey A, Upcroft P, Boreham PFL (1989) Chromosomes of Blastocystis hominis. Int J Parasitol 19:879–883

    Article  CAS  PubMed  Google Scholar 

  • Wu Z, Mirza H, Tan KSW (2014a) Intra-subtype variation in entero adhesion accounts for differences in epithelial barrier disruption and is associated with metronidazole resistance in Blastocystis subtype-7. PLoS Negl Trop Dis 8:e2885

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu Z, Mirza H, Teo JDW, Tan KSW (2014b) Strain-dependent induction of human enterocyte apoptosis by Blastocystis disrupts epithelial barrier and ZO-1 organization in a caspase 3- and 9-dependent manner. Biomed Res Int. https://doi.org/10.1155/2014/209163

    Article  PubMed  PubMed Central  Google Scholar 

  • Yason JA, Tan KSW (2018) Membrane surface features of Blastocystis subtypes. Genes 9:417. https://doi.org/10.3390/genes9080417

    Article  CAS  PubMed Central  Google Scholar 

  • Zaman V, Howe J, Ng M, Goh TK (1999) Scanning electron microscopy of the surface coat of Blastocystis hominis. Parasitol Res 85:974–976

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Zhang S, Qiao J, Wu X et al (2012) Ultra-structural insights into morphology and reproductive mode of Blastocystis hominis. Parasitol Res 110:1165–1172

    Article  PubMed  Google Scholar 

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Acknowledgements

The present work was supported by Ain Shams Faculty of Medicine Grants Office, Grant No. 2016/31.

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Authors and Affiliations

  1. Department of Parasitology, Faculty of Medicine, Ain Shams University, Cairo, Egypt

    Mazloum M. Ahmed, Fayza S. M. Habib, Ghada A. Saad & Heba M. El Naggar

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  1. Mazloum M. Ahmed
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  2. Fayza S. M. Habib
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  3. Ghada A. Saad
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  4. Heba M. El Naggar
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Correspondence to Ghada A. Saad.

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The research protocol was approved by the Ethics Committee, Faculty of Medicine, Ain-Shams University. A written consent from each participant was obtained after providing a clear explanation of the study.

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Ahmed, M.M., Habib, F.S.M., Saad, G.A. et al. Surface ultrastructure, protein profile and zymography of Blastocystis species isolated from patients with colorectal carcinoma. J Parasit Dis 43, 294–303 (2019). https://doi.org/10.1007/s12639-019-01092-9

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  • DOI: https://doi.org/10.1007/s12639-019-01092-9

Keywords

  • Blastocystis sp.
  • Colorectal carcinoma
  • Surface ultrastructure
  • Protein profiles
  • Zymography