Urīna ekzosomu RNS sastāva īpatnības urīnpūšļa vēža pacientiem.
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Date
2021
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Rīgas Stradiņa universitāte
Rīga Stradiņš University
Rīga Stradiņš University
Abstract
Maģistra darba ietvaros analizētas zemas pakāpes urīnpūšļa vēža pacientu urīna ekzosomu sastāva īpatnības. Literatūras apskatā apkopota informācija par urīnpūšļa vēzi un ar to saistītu epidemioloģisku situāciju Latvijā. Mūsdienas lietojamo urīnpūšļa vēza diagnosticēšanas metožu dārgums, invazivitāte un nepietiekami liela jutība rada nepieciešamību pēc jaunas urīnpūšļa vēža diagnosticēšanas un uzraudzības metodes. Par šīs problēmas potenciālo risinājumu tiek uzskatīta ekzosomu kravas specifisko biomarķieru identificēšana, tāpēc šī darba mērķis ir noteikt ekzosomu garo nekodējošo ribonukleīnskābju (lncRNS) sastāva atšķirības starp urīnpūšļa vēža pacientiem un cilvēkiem bez onkoloģiskām saslimšanām. Pētījumam ir Ētikas komitejas atļauja. Pacientu iesaistīšana pētījumā notika Paula Stradiņa Klīniskās universitātes slimnīcas Uroloģijas klīnikā, bet iegūtā bioloģiskā materiāla analizēšana veikta Rīgas Stradiņa universitātes Onkoloģijas institūtā. Tika izveidota pētījuma grupa, kurā tika iekļauti desmit zemas vēža diferenciācijas pakāpes pacienti un gadījumu kontroles grupa, kas sastāv no sešiem pētījuma dalībniekiem. Ar Qiagen polimerāzes ķēdes reakcijas platēm, kas paredz lncRNS identificēšanu un iesaistīto signālceļu atpazīšanu, tika analizētas no urīna ekzosomām izdalītas lncRNS. Iegūto rezultātu statistiska apstrāde tika veikta, izmantojot GeneGlobe Data Analysis Center sistēmu no Qiagen mājaslapas.
Pētījums atklāja, ka salīdzinot pārstāvēto abās grupas lncRNS ekspresiju ar izvēlēto nozīmīguma līmeni (p-vērtība ir 0,05), vēža grupā ir novērota 102 lncRNS samazināta ekspresija, kas ir statistiski ticama (pie kam ar vēzi ir asociētas sekojošas lncRNS: CCAT1, CCAT2, GACAT1, HULC, LUCAT1, MALAT1, PCAT1, PTCSC1, PTCSC3, TUSC7, PCGEM1, UCA1, HEIH, BCAR4, BLACAT1, CAHM, CRNDE, DLEU2, H19, PCA3, PRNCR1, PVT1, RMST) un triju lncRNS paaugstināta ekspresija (LINC00853, TINCR, HAND2-AS1), kas nav statistiski ticama. Par veiktā darba rezultātu ticamību liecina kvalitātes kontroles rezultāti, t.i. negatīva negatīvas genomiskās DNS kontaminācijas kontrole un pozitīva reversās transkripcijas reakcijas pozitīvā kontrole.
Literatūras apskatā apkopota informācija par laika periodu no 1992. līdz 2021. gadam, izmantojot zinātniskās datu bāzes, zinātniskās grāmatas un interneta resursus. Maģistra darbs uzrakstīts latviešu valodā, tā apjoms ir 58 lpp. Darbs satur 16 attēlus, 11 tabulas un tajā izmantots 117 literatūras avots.
The composition peculiarities of urine exosomes of patients with low-grade bladder cancer are analyzed within the scope of the Master Thesis. The literature review summarizes information on bladder cancer and related epidemiological situation in Latvia. Expensiveness, invasiveness, and insufficiently high sensitivity of modern bladder cancer diagnosing methods are creating a necessity in a new bladder cancer diagnosing and monitoring method. The identification of specific biomarkers of exosome cargo is considered as a potential this problem solution, so the aim of this work is to identify differences in the composition of exosomal long non-coding ribonucleic acids (lncRNAs) between bladder cancer patients and people without oncological diseases. The study has the permission of the Ethics Committee. Patient involvement in the study took place in the Urology Clinic of Pauls Stradins Clinical University Hospital, but obtained biological material analysis – in Pauls Stradins Clinical University Institute of Oncology. A research group was created and consisted of ten low-grade patients and a case control group – consisting of six study participants. LncRNAs isolated from urine exosomes were analyzed with Qiagen polymerase chain reaction arrays, which were meant for the identification of lncRNAs and the recognition of the signal pathways involved. The statistical processing of the results was carried out using the GeneGlobe Data Analysis Center system from the Qiagen website. The study found that comparing the lncRNAs expression represented in both groups with the selected level of significance (p-value 0.05), downregulation of 102 lncRNAs was observed in the cancer group, which is statistically significant (following lncRNAs were associated with cancer: CCAT1, CCAT2, GACAT1, HULC, LUCAT1, MALAT1, PCAT1, PTCSC1, PTCSC3, TUSC7, PCGEM1, UCA1, HEIH, BCAR4, BLACAT1, CAHM, CRNDE, DLEU2, H19, PCA3, PRNCR1, PVT1, RMST) and three lncRNAs were upregulated (LINC00853, TINCR, HAND2-AS1), which is not statistically significant. Reliability of this study results is evidenced by the quality control test results: negative genomic DNA contamination controls were negative and positive reverse transcription controls were positive. The literature review summarizes references for the period from 1992 to 2021, including information from scientific databases, scientific monographies and internet resources. Master’s Thesis is written in Latvian, consists of 58 pages. Work contains 16 pictures, 11 tables and 117 references.
The composition peculiarities of urine exosomes of patients with low-grade bladder cancer are analyzed within the scope of the Master Thesis. The literature review summarizes information on bladder cancer and related epidemiological situation in Latvia. Expensiveness, invasiveness, and insufficiently high sensitivity of modern bladder cancer diagnosing methods are creating a necessity in a new bladder cancer diagnosing and monitoring method. The identification of specific biomarkers of exosome cargo is considered as a potential this problem solution, so the aim of this work is to identify differences in the composition of exosomal long non-coding ribonucleic acids (lncRNAs) between bladder cancer patients and people without oncological diseases. The study has the permission of the Ethics Committee. Patient involvement in the study took place in the Urology Clinic of Pauls Stradins Clinical University Hospital, but obtained biological material analysis – in Pauls Stradins Clinical University Institute of Oncology. A research group was created and consisted of ten low-grade patients and a case control group – consisting of six study participants. LncRNAs isolated from urine exosomes were analyzed with Qiagen polymerase chain reaction arrays, which were meant for the identification of lncRNAs and the recognition of the signal pathways involved. The statistical processing of the results was carried out using the GeneGlobe Data Analysis Center system from the Qiagen website. The study found that comparing the lncRNAs expression represented in both groups with the selected level of significance (p-value 0.05), downregulation of 102 lncRNAs was observed in the cancer group, which is statistically significant (following lncRNAs were associated with cancer: CCAT1, CCAT2, GACAT1, HULC, LUCAT1, MALAT1, PCAT1, PTCSC1, PTCSC3, TUSC7, PCGEM1, UCA1, HEIH, BCAR4, BLACAT1, CAHM, CRNDE, DLEU2, H19, PCA3, PRNCR1, PVT1, RMST) and three lncRNAs were upregulated (LINC00853, TINCR, HAND2-AS1), which is not statistically significant. Reliability of this study results is evidenced by the quality control test results: negative genomic DNA contamination controls were negative and positive reverse transcription controls were positive. The literature review summarizes references for the period from 1992 to 2021, including information from scientific databases, scientific monographies and internet resources. Master’s Thesis is written in Latvian, consists of 58 pages. Work contains 16 pictures, 11 tables and 117 references.
Description
Biomedicīna
Biomedicine
Dzīvās dabas zinātnes
Natural Sciences
Biomedicine
Dzīvās dabas zinātnes
Natural Sciences
Keywords
URĪNPŪŠĻA VĒZIS, URĪNS, EKZOSOMAS, GARAS NEKODĒJOŠAS RIBONUKLEĪNSKĀBES, BLADDER CANCER, URINE, EXOSOMES, LONG NON-CODING RIBONUCLEIC ACID