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

Brandi, Jessica; Carlo, Claudia Di; Manfredi, Marcello; Federici, Federica; Bazaj, Alda; Rizzi, Eleonora; Cornaglia, Giuseppe; Laura, Manna; Marengo, Emílio; Cecconi, Daniela

doi:10.1007/s13361-019-02268-6

Cited by 14 publications

(14 citation statements)

References 40 publications

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“…Identification and quantification of proteome modulation of keratinocytes treated with LAB lysates, or exposed to heat-killed S. aureus or S. pyogenes and then treated with LAB lysates, was performed as previously reported 22 . Briefly, cells were collected, washed and lysed in 1X PBS added with protease inhibitors cocktail (Roche) and 0.1% SDS.…”

Section: Methodsmentioning

confidence: 99%

Exploring the wound healing, anti-inflammatory, anti-pathogenic and proteomic effects of lactic acid bacteria on keratinocytes

Brandi

Cheri

Manfredi

et al. 2020

Sci Rep

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The topical application of lactic acid bacteria (LAB) is recognized as a useful approach to improve skin health. This work aims to characterize by a multidisciplinary approach, the wound healing, anti-inflammatory, anti-pathogens and proteomic effects of six LAB lysates, belonging to the genus Lactobacillus. Our results demonstrated that the lysates of tested LAB stimulated the proliferation of keratinocytes, and that L. plantarum SGL 07 and L. salivarius SGL 19 accelerated the re-epithelization by inducing keratinocyte migration. The bacterial lysates also reduced the secretion of specific pro-inflammatory mediators from keratinocytes. Furthermore, viable L. salivarius SGL 19 and L. fermentum SGL 10 had anti-pathogenic effects against S. aureus and S. pyogenes, while L. brevis SGL 12 and L. paracasei SGL 04 inhibited S. aureus and S. pyogenes, respectively. The tested lactobacilli lysates also induced specific proteome modulation of the exposed keratinocytes, involving dysregulation of proteins (such as interleukin enhancer-binding factor 2 and ATP-dependent RNA helicase) and pathways (such as cytokine, NF-kB, Hedgehog, and RUNX signaling) associated with their specific wound healing and anti-inflammatory effects. This study indicates the different potential of selected lactobacilli, suggesting that they may be successfully used in the future together with conventional therapies to bring relief from skin disorders. Abbreviations LAB Lactic acid bacteria TNF-α Tumor necrosis factor-alpha IFN-γ Interferon-gamma PBS Phosphate buffered saline DMEM Dulbecco's modified Eagle's medium MCP-1 Monocyte chemoattractant protein-1 RANTES Regulated on activation, normal T cell expressed and secreted LC-MS/MS Liquid chromatography-tandem mass spectrometry SWATH Sequential window acquisition of all theoretical mass spectra The skin, considered the largest organ of the body, is involved in a variety of functions and acts primarily as a protective barrier preventing the entry of potential pathogens. In particular, skin homeostasis is regulated by microorganisms, the so called skin microbiota, which act on keratinocytes and on their cytokine release, ensuring

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Section: Methodsmentioning

confidence: 99%

Exploring the wound healing, anti-inflammatory, anti-pathogenic and proteomic effects of lactic acid bacteria on keratinocytes

Brandi

Cheri

Manfredi

et al. 2020

Sci Rep

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“…rhamnosus MD 14 was administered, leading to an almost normalization of the colon histology [84]. Similarly, other studies have found lactic bacteria to affect the expression of p53 (refer to Table 1 for specification of bacteria), either mediated through the synthesis of EPS or via non‐specified mechanisms [85–88]. In addition, in vitro studies have shown that n‐butyrate, a common food derivative metabolized by anaerobic bacteria, such as lactobacilli, can induce apoptosis in CRC cell lines by stimulating p21 [10].…”

Section: Carcinoma (Egfr Mtor and P53)mentioning

confidence: 98%

The gut microbiota can orchestrate the signaling pathways in colorectal cancer

Bennedsen

Furbo

Bjarnsholt

et al. 2022

APMIS

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Current evidence suggests that bacteria contribute to the development of certain cancers, such as colorectal cancer (CRC), partly by stimulating chronic inflammation. However, little is known about the bacterial impact on molecular pathways in CRC. Recent studies have demonstrated how specific bacteria can influence the major CRC‐related pathways, i.e., Wnt, PI3K‐Akt, MAPK, TGF‐β, EGFR, mTOR, and p53. In order to advance the current understanding and facilitate the choice of pathways to investigate, we have systematically collected and summarized the current knowledge within bacterial altered major pathways in CRC. Several pro‐tumorigenic and anti‐tumorigenic bacterial species and their respective metabolites interfere with the major signaling pathways addressed in this review. Not surprisingly, some of these studies investigated known CRC drivers, such as Escherichia coli, Fusobacterium nucleatum, and Bacteroides fragilis. Interestingly, some metabolites produced by bacterial species typically considered pathogenic, e.g., Vibrio cholera, displayed anti‐tumorigenic activities, emphasizing the caution needed when classifying healthy and unhealthy microorganisms. The results collectively emphasize the complexity of the relationship between the microbiota and the tumorigenesis of CRC, and future studies should verify these findings in more realistic models, such as organoids, which constitute a promising platform. Moreover, future trials should investigate the clinical potential of preventive modulation of the gut microbiota regarding CRC development.

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“…For proteomics analyses, the data indicated in this paper are extrapolated from an ongoing study on the proteomics and lipidomics analysis of CSCs. In particular, the reported proteome modulations were obtained by a label free approach, as previously described [22]. Briefly, total proteins from Panc1 parental cells and CSCs grown for 2, 4, and 8 weeks were extracted and analyzed by LC-MS/MS using the SWATH method and the differentially expressed proteins were selected according to a statistically significant p-value <0.05 and a fold change threshold of 1.5.…”

Section: Omic Analysesmentioning

confidence: 99%

Progressively De-Differentiated Pancreatic Cancer Cells Shift from Glycolysis to Oxidative Metabolism and Gain a Quiescent Stem State

Ambrosini

Pozza

Fanelli

et al. 2020

Cells

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Pancreatic ductal adenocarcinoma (PDAC) is typically characterized by high chemoresistance and metastatic spread, features mainly attributable to cancer stem cells (CSCs). It is of central interest the characterization of CSCs and, in particular, the study of their metabolic features in order to selectively identify their peculiarities for an efficient therapeutic approach. In this study, CSCs have been obtained by culturing different PDAC cell lines with a specific growth medium. Cells were characterized for the typical stem/mesenchymal properties at short-, medium-, and long-term culture. Metabolomics, proteomics, analysis of oxygen consumption rate in live cells, and the effect of the inhibition of lactate transporter on cell proliferation have been performed to delineate the metabolism of CSCs. We show that gradually de-differentiated pancreatic cancer cells progressively increase the expression of both stem and epithelial-to-mesenchymal transition markers, shift their metabolism from a glycolytic to an oxidative one, and lastly gain a quiescent state. These quiescent stem cells are characterized by high chemo-resistance, clonogenic ability, and metastatic potential. Re-differentiation reverts these features, re-activating their proliferative capacity and glycolytic metabolism, which generally correlates with high aggressiveness. These observations add an important piece of knowledge to the comprehension of the biology of CSCs, whose metabolic plasticity could be exploited for the generation of promising and selective therapeutic approaches for PDAC patients.

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

Cited by 14 publications

References 40 publications

Exploring the wound healing, anti-inflammatory, anti-pathogenic and proteomic effects of lactic acid bacteria on keratinocytes

Exploring the wound healing, anti-inflammatory, anti-pathogenic and proteomic effects of lactic acid bacteria on keratinocytes

The gut microbiota can orchestrate the signaling pathways in colorectal cancer

Progressively De-Differentiated Pancreatic Cancer Cells Shift from Glycolysis to Oxidative Metabolism and Gain a Quiescent Stem State

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