Journal of The American Society for Mass Spectrometry

, Volume 30, Issue 9, pp 1690–1699 | Cite as

Investigating the Proteomic Profile of HT-29 Colon Cancer Cells After Lactobacillus kefiri SGL 13 Exposure Using the SWATH Method

  • Jessica Brandi
  • Claudia Di Carlo
  • Marcello Manfredi
  • Federica Federici
  • Alda Bazaj
  • Eleonora Rizzi
  • Giuseppe Cornaglia
  • Laura Manna
  • Emilio Marengo
  • Daniela CecconiEmail author
Research Article


Despite some studies revealed that kefir acts on different cancers, such as colorectal cancer, the proteomic changes that occur in the colon cancer cells remain to be explored. In this study, the proteomic analysis was combined with determination of kefir characteristics (e.g., adhesion capacity, gastrointestinal and antibiotic resistances), in order to confirm its use as a probiotic. Therefore, a label-free strategy based on SWATH-MS was applied to investigate the proteomic profile of HT-29 cells after exposure for 24 h to a specific strain of Lactobacillus kefiri named SGL 13. We identified a total of 60 differentially expressed proteins in HT-29 cells, among which most are located into the extracellular exosome, playing important/crucial roles in translation and cell adhesion, as indicated by the enrichment analysis. The eIF2 and retinoid X receptor activation pathways appeared to be correlated with the anti-tumoral effect of SGL 13. Immunoblot analysis showed an increase in Bax and a decrease in caspase 3 and mutant p53, and ELISA assay revealed inhibition of IL-8 secretion from HT-29 cells stimulated with LPS upon SGL 13 treatment, suggesting pro-apoptotic and anti-inflammatory properties of kefir. In conclusion, the results of this study, the first of its kind using co-culture of kefir and colon cancer cells, demonstrate that L. kefiri SGL 13 possesses probiotic potency and contribute to elucidate the molecular mechanisms involved in the L. kefiri–colon cancer cell interactions.


SWATH-MS Label-free proteomics Colon cancer Lactobacillus kefiri 



This work was supported by FUR of the MIUR Ministry or Education, University and Research (Italy).

Compliance with Ethical Standards

Conflict of Interest

The authors declared the following potential conflicts of interest with respect of the research, authorship, and/or publication of this article: FF, ER, and LM are employees of Sintal Dietetics s.r.l., Italy, performing all research and development activities for Sintal Dietetics. Sintal Dietetics s.r.l. deposited Lactobacillus kefiri SGL 13 for purposes of European patent.

Supplementary material

13361_2019_2268_MOESM1_ESM.xlsx (116 kb)
Supplemental Table S1 Identified and quantified proteins (XLSX 116 kb)
13361_2019_2268_MOESM2_ESM.xlsx (50 kb)
Supplemental Table S2 Differentially expressed proteins (n = 60). (XLSX 49 kb)
13361_2019_2268_MOESM3_ESM.xlsx (18 kb)
Supplemental Table S3 IPA canonical-enriched pathways. (XLSX 17 kb)


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Copyright information

© American Society for Mass Spectrometry 2019

Authors and Affiliations

  • Jessica Brandi
    • 1
  • Claudia Di Carlo
    • 1
  • Marcello Manfredi
    • 2
    • 3
    • 4
  • Federica Federici
    • 5
  • Alda Bazaj
    • 6
  • Eleonora Rizzi
    • 5
  • Giuseppe Cornaglia
    • 6
  • Laura Manna
    • 5
  • Emilio Marengo
    • 3
    • 7
  • Daniela Cecconi
    • 1
    Email author
  1. 1.Department of Biotechnology, Proteomics and Mass Spectrometry LaboratoryUniversity of VeronaVeronaItaly
  2. 2.ISALIT s.r.l.NovaraItaly
  3. 3.Center for Translational Research on Autoimmune & Allergic Diseases—CAADNovaraItaly
  4. 4.Department of Translational MedicineUniversity of Eastern PiedmontNovaraItaly
  5. 5.Sintal Dietetics s.r.l.TeramoItaly
  6. 6.Department of Diagnostics and Public HealthUniversity of VeronaVeronaItaly
  7. 7.Department of Sciences and Technological InnovationUniversity of Eastern PiedmontAlessandriaItaly

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