Izražanje genov mitomiR in Mct1 ter vpliv na metabolizem celic mišjega mamarnega adenokarcinoma

Radanović, Nada (author), Čemažar, Maja (mentor), Renčelj, Andrej (co-mentor)

MitomiR so molekule mikroRNA (miRNA), ki so prisotne predvsem v mitohondrijih. Novejše študije so dokazale, da mitomiR uravnavajo več onkogenih signalnih poti in ciljajo na ključne prenašalce in encime v celični presnovi. Poleg tega mitomiR nadzorujejo presnovo rakavih celic preko uravnavanja izražanja informacijske ribonukleinske kisline (mRNA). Monokarboksilatni transporter 1 (MCT1) je pomemben transportni protein, ki je udeležen pri transportu monokarboksilatov, predvsem L-laktata, ketonskih telesc in piruvata preko membrane. Študije kažejo, da je izražanje gena Mct1 v nekaterih vrstah raka, kot je rak dojke, močno povišano. Namen našega dela je bil primerjati stopnjo izražanja miR-181c-5p, miR-124-3p in gena Mct1 med kontrolno skupino celic TS/A mišjega mamarnega adenokarcinoma ter skupinami celic, ki so bile tretirane z anti-miR. Anti-miR so kemično modificirane molekule RNA, zasnovane za specifično vezavo na molekule miRNA in njihovo utišanje. Poleg tega smo želeli ugotoviti razlike v vrednosti intracelularnega pH (pHi), aktivnosti citokrom c oksidaze ter rasti in preživetju med kontrolnimi celicami TS/A in celicami, tretiranimi z anti-miR. Izražanje izbranih mitomiR in gena Mct1 smo kvantificirali s pomočjo kvantitativne verižne reakcije polimeraze v realnem času (qRT-PCR). Določili smo razlike v vrednosti pHi in aktivnosti citokrom c oksidaze v celicah TS/A. Preživetje in rast celic TS/A smo spremljali s pomočjo testa PrestoBlueTM. Ugotovili smo, da celice TS/A mišjega mamarnega adenokarcinoma izražajo miR-181c-5p, miR-124-3p in gen Mct1, vendar nismo dokazali, da se po tretiranju izbranih mitomiR spremeni izražanje gena Mct1. Dokazali smo, da se z tretiranjem izbranih mitomiR spremenita vrednost pHi in aktivnost citokrom c oksidaze, vendar rast in preživetje celic TS/A ostajata nespremenjena.

• Language: Slovenian
• Keywords: biologija, adenokarcinom, metabolizem, mitomiR, monokarboksilatni transporter 1
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [N. Radanović]
• Year: 2023
• PID: 20.500.12556/RUL-152018
• UDC: 618.19-006(043.2)
• COBISS.SI-ID: 170632451
• Publication date in RUL: 28.10.2023

Secondary language

• Language: English
• Title: The expression level of selected mitomiR and Mct1 and effect on the metabolism of mouse mammary adenocarcinoma cells

Abstract

MitomiRs are microRNA (miRNA) molecules primarily present in mitochondria. Recent studies have shown that mitomiRs regulate multiple oncogenic signaling pathways and target key transporters and enzymes in cellular metabolism. In addition, mitomiRs control the metabolism of cancer cells by regulating the expression of messenger ribonucleic acid (mRNA). Monocarboxylate transporter 1 (MCT1) is an important transport protein involved in the transport of monocarboxylates, especially L-lactate, ketone bodies and pyruvate across the membrane. Studies have shown that the expression of the Mct1 gene is greatly increased in some types of cancer, such as breast cancer. The aim of our study was to compare the expression levels of miR-181c-5p, miR-124-3p and the Mct1 gene between the control group of TS/A mouse mammary adenocarcinoma cells and groups of cells treated with anti-miRs. Anti-miRs are chemically modified RNA molecules designed for specific binding to miRNA molecules and their silencing. Additionaly, we wanted to determine differences in intracellular pH (pHi) values, cytochrome c oxidase activity, as well as cell growth and survival between control cells and cells treated with anti-miRs. The expression of selected mitomiRs and the Mct1 gene was quantified using quantitative real-time polymerase chain reaction (qRT-PCR). We determined differences in pHi values and cytochrome c oxidase activity in TS/A cells. The survival and growth of TS/A cells were monitored using the PrestoBlueTM assay. We have demonstrated that TS/A mouse mammary adenocarcinoma cells express miR-181c-5p, miR-124-3p and Mct1 gene, but we did not demonstrate changes in the expression of the Mct1 gene after treating the selected mitomiRs. We have shown that treating of selected mitomiRs changes the values of pHi and cytochrome c oxidase activity, but does not change the growth and survival of TS/A cells.

• Keywords: biology, adenocarcinoma, metabolism, mitomiRs, monocarboxylate transporter 1