Title Citotoksično djelovanje različitih varijeteta Raphanus sativus na stanične linije humanih karcinoma mjereno MTT metodom
Title (english) The cytotoxic effect of different varieties of Raphanus sativus on different human cancer cell lines measured by MTT method
Author Anamarija Bilić
Mentor Vedrana Čikeš Čulić (mentor)
Committee member Vedrana Čikeš Čulić (predsjednik povjerenstva)
Committee member Marina Degoricija (član povjerenstva)
Committee member Ana Šešelja Perišin (član povjerenstva)
Granter University of Split School of Medicine (Medical chemistry and biochemistry) Split
Defense date and country 2021-10-14, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Pharmacy Medical Biochemistry
Abstract Cilj istraživanja:
Cilj ovog istraživanja je ispitati potencijalno citotoksično djelovanje ekstrakata biljnih vrsta Raphanus sativus (L.) i Brassica rapa (L.) na humane karcinomske stanice dojke MDA-MB-231 te mokraćnog mjehura T24. Pretpostavka je da će se broj karcinomskih stanica nakon izlaganja ekstraktima smanjiti u odnosu na kontrolnu skupinu.
Materijali i metode:
Citotoksičnost na staničnim linijama karcinoma dojke i mokraćnog mjehura ispitivala se MTT metodom. Njome se određuje postotak metabolički aktivnih stanica nakon tretiranja s mikrovalnim ekstraktima biljnih vrsta Raphanus sativus (L.) i Brassica rapa (L.) Uzorci biljki pripravljeni su u različitim koncentracijama, a učinak se određivao nakon 4, 24, 48 i 72 sata. Djelotvornost ekstrakata određivana je spektrofotometrijski mjerenjem apsorbancije pri 570 nm. Uspoređivanjem apsorbancije nastalog formazana kod stanica tretiranih uzorcima biljki i apsorbancije kod kontrole (netretirnih stanica), dobili smo postotak preživljenja karcinomskih stanica.
Rezultati:
Rezultati su prikazani grafički u odnosu vremena inkubacije i postotka metabolički aktivnih stanica.
Kod stanica karcinoma dojke, većina uzoraka je najveći citotoksični učinak imala pri koncentracijama od 50 µg/mL, 75 µg/mL i 100 µg/mL nakon 72 h inkubacije. Najznačajniji učinak postignut je kod uzorka crne rotkvice, dobivenog mikrovalnom ekstrakcijom, pri koncentraciji od 100 µg/mL, gdje se nakon 72 h broj metabolički aktivnih stanica smanjio na 48%.
Kod stanica karcinoma mokraćnog mjehura, većina uzoraka je najveći citotoksični učinak imala pri višim koncentracijama nakon 48 i 72 h inkubacije.
Najznačajniji učinak postignut je kod uzorka bijele repe, dobivenog mikrovalnom ekstrakcijom, pri koncentraciji od 50 µg/mL, gdje se nakon 72 h broj metabolički aktivnih stanica smanjio na 16%.
Zaključak:
In vitro izlaganje stanica karcinoma dojke MDA-MB-231 i karcinoma mokraćnog mjehura T24 uzorcima crne rotkvice i bijele repe, dobivenih mikrovalnom ekstrakcijom, dovodi do smanjenja preživljenja tih stanica. Ispitivani ekstrakti pokazuju citotoksični učinak ovisan o vremenu inkubacije i koncentraciji. Djelovanje ekstrakata nije uvijek razmjerno povećanju vremena inkubacije i koncentracije te u pojedinim slučajevima dolazi do oporavka stanica. Najveći citotoksični učinak na staničnu liniju MDA-MB-231 ostvario je mikrovalni ekstrakt crne rotkvice nakon 72 h inkubacije, a na staničnu liniju T24 ekstrakt bijele repe nakon 72 h inkubacije. Citotoksični učinak ekstrakata Raphanus sativus (L.) i Brassica rapa (L.), što je ujedno i hipoteza našeg ispitivanja, je potvrđen. Idući korak bio bi potvrđivanje tih učinaka in vivo ispitivanjem na modelima karcinoma dojke i mokraćnog mjehura kod životinja.
Abstract (english) The aim of the research:
The aim of this research is to examine the potential cytotoxic effects of plant species extracts of Raphanus sativus (L.) and Brassica rapa (L.) on human carcinoma cells of breast MDA-MB-231 and bladder T24. It is assumed that the number of cancer cells will decrease after exposure to the extracts in comparison to the control group.
Materials and methods:
The cytotoxicity on breast and bladder cancer cell lines was tested by the MTT method. It is used to determine the percentage of metabolically active cells after exposure to microwave extracts of plant species Raphanus sativus (L.) and Brassica rapa (L.). Plant samples were prepared in different concentrations and the effect is seen after 4, 24, 48 and 72 hours. The efficiency of the extracts was observed spectrophotometrically by measuring the absorbance at 570 nm. By comparing the absorbance of the formed formazan in cells treated with plant samples and the absorbance of the control (non-treated cells), we obtained the survival rate of cancer cells.
Results:
The results are graphically presented in relation to the incubation time and the percentage of metabolically active cells.
In breast cancer cells, most samples had the greatest cytotoxic effect at concentrations of 50 μg/mL, 75 μg/mL and 100 μg/mL after 72 h of incubation. The most significant effect was achieved with the black radish sample, obtained by microwave extraction, at a concentration of 100 µg/mL, where after 72 h the number of metabolically active cells decreased to 48%.
In bladder cancer cells, most samples had the greatest cytotoxic effect at higher concentrations after 48 and 72 h of incubation.
The most significant effect was achieved with the white turnip sample, obtained by microwave extraction, at a concentration of 50 µg/mL, where after 72 h the number of metabolically active cells decreased to 16%.
Conclusions:
In vitro exposure of breast cancer cells MDA-MB-231 and bladder cancer T24 to samples of black radish and white turnip, obtained by microwave extraction, leads to decrease in the survival of these cells. The tested extracts show a cytotoxic effect depended on the incubation time and concentration. Effect of extracts is not always proportional to the increase in incubation time and concentration and in some cases cell recovery occurs. The greatest cytotoxic effect on the MDA-MB-231 cell line was achieved by black radish microwave extract after 72 h of incubation and on T24 cell line by white turnip extract after 72 h of incubation. The cytotoxic effect of the extracts of Raphanus sativus (L.) and Brassica rapa (L.), which was also the hypothesis of our research, was confirmed. The next step is the confirmation of these effects by in vivo studies on the models of animal breast and bladder cancers.
Keywords
Rotkvica
Bijela repa
Ljekovito bilje-toksičnost
Tumorske stanične linije
Novotvorine dojki
Novotvorine mokraćnog mjehura
Testovi probira antitumorskih lijekova
Keywords (english)
Raphanus
Brassica rapa
Plants Medicinal-toxicity
Cell Line Tumor
Breast Neoplasms
Urinary Bladder Neoplasms
Drug Screening Assays Antitumor
Language croatian
URN:NBN urn:nbn:hr:171:960897
Project Number: IP-2016-06-1316 Title: Biljke kao izvor bioaktivnih sumporovih spojeva te njihova sposobnost hiperakumulacije metala Title: Plants as a source of bioactive sulphur compounds and their ability to hyperaccumulate metals Acronym: BioSMe Leader: Ivica Blažević Jurisdiction: Croatia Funder: HRZZ Funding stream: IP
Study programme Title: Pharmacy Study programme type: university Study level: integrated undergraduate and graduate Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
Type of resource Text
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Created on 2021-11-05 10:04:33