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Title:Vpliv izbranih UV-filtrov na proliferacijsko aktivnost melanomskih celic ter na izražanje mRNA za transmembranski protein ABCB5
Authors:ID Prunk Zdravković, Tanja (Author)
ID Ferk, Polonca (Mentor) More about this mentor... New window
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Language:Slovenian
Work type:Doctoral dissertation
Organization:MF - Faculty of Medicine
Abstract:UVOD: Oktokrilen (OCT) in titanijev (IV) oksid (TiO2) sta ena izmed najpogosteje uporabljenih UV-filtrov v kremah za sončenje in drugih izdelkih za osebno nego. Trenutno še ni jasnih dokazov o tem, ali delujejo zaščitno proti melanomu ali lahko morda celo predstavjajo dejavnik tveganja pri razvoju te najnevarnejše in najbolj agresivne oblike kožnega raka. Zato je bil cilj te raziskave preučiti morebiten vpliv OCT in TiO2 (v nanodelcih in delcih mikronskih velikosti; nano-TiO2 in mikro-TiO2) na proliferacijsko aktivnost, citotoksičnost in izražanje mRNA za prenašalec ABCB5, ki ga melanomske celice izražajo. Transmembranski protein ABCB5 smo testirali zaradi njegove znane vloge v iniciaciji, lokalni invaziji in metastaziranju različnih vrst raka, vključno z melanomom. METODE: metastatsko melanomsko celično linijo WM-266-4 (ATCC) smo izpostavili izbranim koncentracijam OCT, nano-TiO2 in mikro-TiO2 v različnih časovnih intervalih (2 h, 8 h, 24 h, 48 h, 120 h, 144 h). Proliferacijsko aktivnost smo merili s testom MTT in citotoksičnost s testom LDH. Izražanje tarčnega gena (ABCB5) smo detektirali s kvantitativnim PCR v realnem času (RT-qPCR) z uporabo sond TaqMan®. REZULTATI: Naši rezultati nakazujejo na celokupno zmanjšano proliferativno aktivnost, citotoksičen vpliv ter povečano izražanje mRNA za prenašalec ABCB5 (p < 0,05) pri metastatskih melanomskih celicah po izpostavitvi OCT in nano-TiO2. Z daljšim časom izpostavljenosti in z večjo koncentracijo filtra v primerjavi s kontrolnimi celicami so rezultati bolj značilni. Zaradi tega domnevamo, da bi preživele celice lahko bile mnogo bolj invazivne in agresivne, kar lahko pojasni tudi mikroskopsko ugotovljen celični kanibalizem. Mikro-TiO2 pri isti celični liniji ima prav tako citotoksičen vpliv, vendar z daljšim časom izpostavljenosti v preživelih celicah pride do zmanjšanega izražanja mRNA za prenašalec ABCB5. ZAKLJUČEK: Naši rezultati nakazujejo, da za paciente z diagnosticiranim metastatskim melanomom ni priporočljivo uporabljati izdelkov, ki vsebujejo OCT ali nano-TiO2, saj to lahko pospeši napredovanje bolezni, okrepi kemorezistenco in vodi do ponovitve bolezni. V nasprotju s tema filtroma, pa bi uporaba izdelkov z mikro-TiO2 pri teh pacientih lahko bila varna. Nadaljnje raziskave bodo morale okrepiti rezultate, pridobljene s to študijo, s preučevanjem zdrave celične kulture človeških melanocitov ter primarne melanomske človeške celične linije. Ta doktorska disertacija odpira novo obetavno raziskovalno področje, z namenom bolje osvetliti etiologijo in prispevati k boljši preventivi melanoma.
Keywords:oktokrilen, titanijev (IV) oksid nano in mikro velikosti, melanom, protein ABCB5
Place of publishing:Maribor
Year of publishing:2018
PID:20.500.12556/DKUM-69983 New window
COBISS.SI-ID:298372096 New window
NUK URN:URN:SI:UM:DK:EOMWXCB8
Publication date in DKUM:26.02.2019
Views:1942
Downloads:0
Metadata:XML DC-XML DC-RDF
Categories:MF
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Licences

License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:22.03.2018

Secondary language

Language:English
Title:The influence of selected UV-filters on the proliferation activity of melanoma cells and on their ABCB5 transmembrane protein mRNA expression
Abstract:INTRODUCTION: Octocrylene (OCT) and Titanium (IV) Oxide (TiO2) are one of the most widespread UV-filters used in sunscreens and personal care products. There is no clear evidence on whether they are protective against or may be a contributing factor in melanoma development. Hence the aim of this study was to investigate any potential influence of OCT and TiO2 (in nanoparticles and microparticles; nano-TiO2 and micro-TiO2) on proliferative activity, cytotoxicity and ABCB5 mRNA expression in melanoma cells. The ABCB5 transmembrane protein was tested due to its well-known role in the initiation, invasion and metastatic spread of various cancers, including melanoma. METHODS: Metastatic melanoma cell line WM-266-4 (ATCC) was incubated with selected concentrations of OCT and TiO2 for different time intervals. The MTT and LDH assays to measure the cells' proliferative activity and cytotoxicity were used respectively. Target gene (ABCB5) expression was detected by quantitative real-time PCR (qRT-PCR), using TaqMan® chemistry. RESULTS: Our results suggest decreased metastatic melanoma cells’ proliferative activity, increased cytotoxicity and increased ABCB5 mRNA expression (p < 0.05) with longer time of exposure to and higher concentrations of OCT and nano-TiO2 as compared to control cells. Accordingly, we suspect that the surviving cells are more invasive and aggressive, which might explain their microscopically observed cannibalistic activity. Micro-TiO2 in the same cell line also has a cytotoxic effect, but with a longer exposure time in the surviving cells, the expression of mRNA for the ABCB5 protein is decreased. CONCLUSIONS: Our results suggest that it is not advisable to use products containing OCT or nano-TiO2 for patients with diagnosed metastatic melanoma, as this can accelerate disease progression, strengthen chemoresistence and lead to a recurrence of the disease. In contrast to these filters, the use of micro-TiO2 products in these patients could be safe. Further research will need to strengthen the results obtained by this study by studying the healthy cell culture of human melanocytes and the primary melanoma human cell line. With this doctoral thesis, we elucidate a new promising field for further research to contribute to etiology and prevention of melanoma.
Keywords:octocrylene, titanium (IV) oxide nanoparticles and microparticles, melanoma, ABCB5 protein


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