In vitro ispitivanje utjecaja S. enterica LT21 na metabolizam i ekspresiju proteina HEp2 stanica

master's thesis

• Title (english): In vitro investigation of influence of S. enterica LT21 on metabolism and protein expression of HEp2 cells
• Author: Jelena Miđić
• Mentor: Ksenija Durgo (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Biology and Microbial Genetics); Zagreb
• Defense date and country: 2014-09-19, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

S. enterica stvara mnogobrojne probleme kad dođe do kontaminacije ljudskog organizma ovom bakterijom. L. plantarum ima mnogobrojne pozitivne učinke na ljudsko zdravlje. U ovom radu ispitan je utjecaj enterobakterije S. enterica LT21 na humanu staničnu liniju karcinoma grkljana (HEp2). Ispitano je svojstvo adhezije bakterija S. enterica LT21 kao samostalne kulture i u kombinaciji s L. plantarum 16.4 na HEp2 stanice. Također, istražena je metabolička aktivnost bakterije S. enterica LT21 nakon adhezije na HEp2 stanice kao i sadržaj proteina u HEp2 stanicama nakon tretmana s bakterijom S. enterica LT21. S. enterica LT21 se bolje veže nego L. plantarum 16.4 kad su bakterije pomiješane u omjerima 1:1 i 10:1. Analizom organskih kiselina detektirane su octena i mliječna kiselina u uzorku HEp2 stanica inkubiranih sa S. enterica LT21, te mliječna kiselina u uzorku HEp2 stanica. Proteomska ispitivanja provedena su pomoću analize tandemne spektrometrije masa (MS/MS). S. enterica LT21 utječe na ekspresiju proteina HEp2 stanica. Nakon tretiranja HEp2 stanica sa S. enterica LT21 dodatno je eksprimiran protein troponin T tipa 1, a reprimirana je ekspresija četiri proteina: tioredoksin, galektin-1 u kompleksu s mliječnom kiselinom, destrin i S-adenozilhomocistein hidrolaza. Neki od reprimiranih proteina bitni su za preživljavanje tumorskih stanica.

Abstract (english)

S. enterica is a pathogen that causes health problems. On the other hand, L. plantarum has numerous positive effects on human health. The aim of this study was to investigate the influence of enterobacteria S. enterica LT21 on human laryngeal carcinoma cell line (HEp2). The ability of the adhesion of S. enterica LT21 as an independent culture and in combination with L. plantarum 16.4 to HEp2 cells was tested as well as metabolic activity of S. enterica LT21 after adhesion to HEp2 cells. The protein content in HEp2 cells after the incubation with S. enterica LT21 was invastigated and it was compared with the protein content in the untreated HEp2 cells. Adhesion results show better binding of S. enterica LT21 than L. plantarum 16.4 when bacteria are mixed in a 1:1 and 10:1 ratio. Acetic and lactic acid in the sample of HEp2 cells incubated with S. enterica LT21 were detected, as well as lactic acid in a sample of HEp2 cells. Proteomic studies were performed using tandem mass spectrometry analysis (MS/MS). S. enterica LT21 affects the expression of proteins of HEp2 cells. HEp2 cells after incubation with S. enterica LT21 additionally expressed protein troponin T type 1, and repressed the expression of four proteins: thioredoxin, galectin-1 in complex with lactic acid, destrin and S-adenosylhomocystein hydrolase. Some of repressed proteins are essential for the survival of tumor cells.

• Keywords: S. enterica LT21; HEp2; adhezija; proteomika; metabolizam
• Keywords (english): S. enterica LT21; HEp2; adhesion; proteomics; metabolism
• Language: croatian
• URN:NBN: urn:nbn:hr:159:916824
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-05-02 12:35:05