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FOXC1-Mediated Effects of miR-204-5p on Melanoma Cell Proliferation / I. Y. Dubovtseva, M. B. Aksenenko, E. D. Nikolaeva [et al.] // Mol. Biol. - 2021. - Vol. 55, Is. 4. - P610-617, DOI 10.1134/S0026893321020199. - Cited References:24 . - ISSN 0026-8933. - ISSN 1608-3245

РУБ Biochemistry & Molecular Biology
Рубрики:
FOXC1
Кл.слова (ненормированные):
FOXC1 -- miR-204-5p -- melanoma -- BRO -- SK-MEL-2 -- siRNA -- miRNA -- dormant cancer -- cells
Аннотация: MicroRNAs epigenetically regulate physiological and pathological processes. Previously, we found that miR-204-5p is expressed at low levels in melanoma cells, and an increase in its level leads to a change in proliferation, migration, and invasion of these cancer cells. Now, using bioinformatics analysis, it has been shown that the target of miR-204-5p is FOXC1 transcription factor, which is implicated in carcinogenesis. Using the luciferase reporter assay, it was found that miR-204-5p suppresses expression of the FOXC1 gene by binding to its 3' non-coding region. Transfection of small interfering RNA (siRNA) targeting FOXC1 into melanoma cells caused a decrease in miR-204-5p levels, which is consistent with the generally accepted concept of feedback regulation of miRNA expression by target genes. According to the results of the MTT test and fluorescence microscopy, the proliferation level of melanoma cells under the influence of siRNA to FOXC1 decreased 72 h after transfection. Changes in the ratio of cells by cell cycle phase were analyzed using flow cytometry. Regulatory relationships between FOXC1 and miR-204-5p, and an inhibitory effect of FOXC1 knockdown on melanoma cell proliferation were revealed. Based on the results, it can be assumed that miR-204-5p regulates proliferation of melanoma cells by affecting FOXC1 expression.

WOS
Держатели документа:
Voino Yasenetsky Krasnoyarsk State Med Univ, Minist Hlth Russian Federat, Krasnoyarsk 660022, Russia.
RAS, Biophys Inst, Siberian Branch, Div Fed Res Ctr,Krasnoyarsk Sci Ctr, Krasnoyarsk 660022, Russia.
Russian Acad Sci, Siberian Branch, Res Inst Med Problems North, Krasnoyarsk 660022, Russia.

Доп.точки доступа:
Dubovtseva, I. Yu; Aksenenko, M. B.; Nikolaeva, E. D.; Averchuk, A. S.; Moshev, A., V; Savchenko, A. A.; Markova, S., V; Ruksha, T. G.

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FOXC1-Mediated Effects of miR-204-5p on Melanoma Cell Proliferation / I. Y. Dubovtseva, M. B. Aksenenko, E. D. Nikolaeva [и др.] // Mol Biol (Mosk). - 2021. - Vol. 55, Is. 4. - С. 667-675, DOI 10.31857/S0026898421030058 . - ISSN 0026-8984
Кл.слова (ненормированные):
BRO -- dormant cancer cells -- FOXC1 -- melanoma -- miR-204-5p -- miRNA -- siRNA -- SK-MEL-2 -- forkhead transcription factor -- FOXC1 protein, human -- microRNA -- MIRN204 microRNA, human -- cell motion -- cell proliferation -- genetics -- human -- melanoma -- tumor cell line -- Cell Line, Tumor -- Cell Movement -- Cell Proliferation -- Forkhead Transcription Factors -- Humans -- Melanoma -- MicroRNAs
Аннотация: MicroRNAs epigenetically regulate physiological and pathological processes. Previously, we found that miR-204-5p is expressed at low levels in melanoma cells, and an increase in its level leads to a change in proliferation, migration, and invasion of these cancer cells. Now, using bioinformatics analysis, it has been shown that the target of miR-204-5p is FOXC1 transcription factor, which is implicated in carcinogenesis. Using the luciferase reporter assay, it was found that miR-204-5p suppresses expression of the FOXC1 gene by binding to its 3' non-coding region. Transfection of small interfering RNA (siRNA) targeting FOXC1 into melanoma cells caused a decrease in miR-204-5p levels, which is consistent with the generally accepted concept of feedback regulation of miRNA expression by target genes. According to the results of the MTT test and fluorescence microscopy, the proliferation level of melanoma cells under the influence of siRNA to FOXC1 decreased 72 h after transfection. Changes in the ratio of cells by cell cycle phase were analyzed using flow cytometry. Regulatory relationships between FOXC1 and miR-204-5p, and an inhibitory effect of FOXC1 knockdown on melanoma cell proliferation were revealed. Based on the results, it can be assumed that miR-204-5p regulates proliferation of melanoma cells by affecting FOXC1 expression.

Scopus
Держатели документа:
Voino-Yasenetsky Krasnoyarsk State Medical University, Ministry of Health of the Russian Federation, Krasnoyarsk, 660022, Russian Federation
Siberian Branch of the Russian Academy of Sciences, Research Institute for Medical Problems in the North, Krasnoyarsk, 660022, Russian Federation
Biophysics Institute of the Siberian Branch of the RAS - Division of Federal Research Center "Krasnoyarsk Scientific Center of the Siberian Branch of the RAS", Krasnoyarsk, 660022, Russian Federation

Доп.точки доступа:
Dubovtseva, I. Y.; Aksenenko, M. B.; Nikolaeva, E. D.; Averchuk, A. S.; Moshev, A. V.; Savchenko, A. A.; Markova, S. V.; Ruksha, T. G.

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Large-scale, high-throughput validation of short hairpin RNA sequences for RNA interference [Text] / L. H. Lamarcq [et al.] // J. Biomol. Screen. - 2006. - Vol. 11, Is. 3. - P236-246, DOI 10.1177/1087057105284342. - Cited References: 50 . - ISSN 1087-0571

РУБ Biochemical Research Methods + Biotechnology & Applied Microbiology + Chemistry, Analytical
Рубрики:
ENZYME FRAGMENT COMPLEMENTATION
   ORFEOME VERSION 1.1
   SMALL NUCLEAR-RNA
   MAMMALIAN-CELLS
   GENE-EXPRESSION
   SIRNA SEQUENCES
   POLYMERASE-III
   SELECTION
   TRANSCRIPTION
   MICROARRAYS
Кл.слова (ненормированные):
RNAi -- high-throughput screening -- target validation -- shRNA -- reporter assay
Аннотация: (shRNAs) is described. Using this approach, 464 shRNAs auainst 116 different genes were screened for knockdown efficacy, enabling rapid identification of effective shRNAS against 74 genes. Statistical analysis of the effects of various criteria on the activity of the shRNAs confirmed that some of the rules thought to govern small interfering RNA (siRNA) activity also apply to shRNAs. These include moderate GC content, absence of internal hairpins, and asymmetric thermal stability. However, the authors did not find strong support for position-specific rules. In addition, analysis of the data suggests that not all genes are equally susceptible to RNA interference (RNAi).

Держатели документа:
Clontech Labs, Mountain View, CA 94043 USA
BD Biosci Pharmingen, La Jolla, CA USA
Bayer Healthcare AG, Pharma Res, Mol Screening Technol, Wuppertal, Germany
Russian Acad Sci, Inst Biophys, Photobiol Lab, Siberian Branch, Krasnoyarsk, Russia
ИБФ СО РАН : 660036, Красноярск, Академгородок, д. 50, стр. 50

Доп.точки доступа:
Lamarcq, L.H.; Scherer, B.J.; Phelan, M.L.; Kalnine, N.N.; Nguyen, Y.H.; Kabakova, T...; Chen, X.Y.; Tan, M...; Chang, C...; Berlon, C...; Campos-Gonzalez, R...; Gao, G.J.; Golz, S...; Vysotski, E.S.; Farmer, A.A.

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