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A bioluminescent assay for detecting melanocortin-1 receptor (MC1R) gene polymorphisms R160W, R151C, and D294H / E. E. Bashmakova [et al.] // Mol. Biol. - 2015. - Vol. 49, Is. 6. - P852-857, DOI 10.1134/S0026893315050039 . - ISSN 0026-8933
Кл.слова (ненормированные):
bioluminescent assay -- MC1R receptor -- melanoma -- single nucleotide polymorphisms
Аннотация: Several polymorphisms in the melanocortin-1 receptor gene (MC1R) have been associated with melanoma risk. In particular, rs1805007, rs1805008, and rs1805009 mutations, which result in R151C, R160W, and D294H amino acid substitutions, respectively, and are associated with the phenotype of red-hair mutations, have also been connected with melanoma and nonmelanoma skin cancer risks. This work describes a method of detecting these polymorphisms using primer extension with subsequent dual bioluminescent assay. Model plasmids carrying polymorphic MC1R fragments, as well as several clinical DNA samples, were tested using the proposed technique. The results agreed well with those obtained by Sanger sequencing. © 2015, Pleiades Publishing, Inc.

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Держатели документа:
Siberian Federal University, Krasnoyarsk, Russian Federation
Blokhin Cancer Research Center, Moscow, Russian Federation
Institute of Biophysics, Siberian Branch of the Russian Academy of Sciences, Krasnoyarsk, Russian Federation
Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russian Federation
Voino-Yasenetskii Krasnoyarsk State Medical University, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Bashmakova, E. E.; Krasitskaya, V. V.; Bondar, A. A.; Kozlova, A. V.; Ruksha, T. G.; Frank, L. A.
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Bioluminescent SNP genotyping technique: Development and application for detection of melanocortin 1 receptor gene polymorphisms / E. E. Bashmakova [et al.] // Talanta. - 2018. - Vol. 189. - P111-115, DOI 10.1016/j.talanta.2018.06.057 . - ISSN 0039-9140
Кл.слова (ненормированные):
Ca2+-regulated photoprotein obelin -- Genotyping -- Melanocortin 1 receptor gene -- Single nucleotide polymorphisms (SNP) -- Bioluminescence -- Clinical research -- Curricula -- Diagnosis -- Genes -- Oncology -- Biomedical research -- Clinical characteristics -- Development and applications -- Genotyping -- Healthy individuals -- Photoproteins -- Receptor genes -- Single-nucleotide polymorphisms -- Dermatology
Аннотация: SNP genotyping based on the reaction of specific primer extension with the following bioluminescent detection of its products was shown to be potentially applicable for biomedical exploration. The paper describes its elaboration and first application in extensive biomedical research concerning MC1R gene variants’ frequency and associations with clinical characteristics in melanoma patients of Eastern Siberia (Krasnoyarsk region, Russia). Polymorphisms rs 1805007 (R151C), rs 1805008 (R160W), and rs 1805009 (D294H) were detected in 174 DNA samples from patients with histologically proved diagnosis of cutaneous melanoma and in 200 samples from healthy individuals. All the results on bioluminescent SNP genotyping were confirmed by Sanger sequencing. Some features characteristic of the population were found, i.e. melanoma is mostly associated with R160W or R151C while variant D294H is extremely rare; simultaneous carriage of any two investigated variants is also strongly associated with melanoma; R151C is associated with ulceration and consequently the disease course is more aggressive, etc. The design of the technique allows fast evaluation of any known diagnostically important SNP frequencies and associations across population. © 2018 Elsevier B.V.

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Держатели документа:
Siberian Federal University, Svobodny pr. 79, Krasnoyarsk, Russian Federation
Institute of Biophysics SB RAS, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Akademgorodok 50/50, Krasnoyarsk, Russian Federation
Blokhin Cancer Research Center, Moscow, Russian Academy of Medical Sciences, Kashirskoye Shosse 24, Moscow, Russian Federation
Institute of Chemical Biology and Fundamental Medicine, SB RAS, Novosibirsk Lavrentiev Avenue 8, Novosibirsk, Russian Federation
State Medical University named after V.F. Voyno-Yasenetsky, Partizana Zheleznyaka St. 1, Krasnoyarsk, Russian Federation
Regional Clinical Oncology Center named after A.I. Kryzhanovsky, 1 Smolenskaya Str.16, Krasnoyarsk, Russian Federation

Доп.точки доступа:
Bashmakova, E. E.; Krasitskaya, V. V.; Bondar, A. A.; Eremina, E. N.; Slepov, E. V.; Zukov, R. A.; Frank, L. A.

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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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N-extended photoprotein obelin to competitively detect small protein tumor markers / E. E. Bashmakova, N. S. Panamarev, A. N. Kudryavtsev, L. A. Frank // Biochem. Biophys. Res. Commun. - 2022. - Vol. 598. - P69-73, DOI 10.1016/j.bbrc.2022.02.011. - Cited References:15. - The work was partially supported by a grant of the President of the Russian Federation for young scientists, the candidates of sciences (project MK-772.2020.4, study of a hybrid protein with melanoma-inhibiting activity) and Government of Krasnoyarsk Territory, Krasnoyarsk Regional Science Foundation (project No 2021012006966, study of a hybrid with protein survivin). . - ISSN 0006-291X. - ISSN 1090-2104

РУБ Biochemistry & Molecular Biology + Biophysics
Рубрики:
MELANOMA INHIBITORY-ACTIVITY
SURVIVIN
Кл.слова (ненормированные):
Photoprotein obelin -- Genetic fusion -- Tumor marker -- Competitive assay
Аннотация: Two variants of Ca2+-regulated photoprotein obelin, extended from the N-terminus with small tumor markers-melanoma inhibitory activity protein (MIA) and survivin, one of the protein inhibitors of apoptosis, were designed, obtained and studied. Both domains in the obtained hybrid proteins exhibit the properties of the initial molecules: the main features of Ca2+-triggered bioluminescence are close to those of obelin, and the tumor markers' domains are recognized and bound by the corresponding antibodies. The obtained hybrids compete with the corresponding tumor markers for binding with antibodies, immobilized on the surface and their use has been shown to be promising as bioluminescent labels in a one-stage solid-phase competitive immunoassay. (c) 2022 Published by Elsevier Inc.

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Держатели документа:
Fed Res Ctr Krasnoyarsk Sci Ctr SB RAS, Inst Biophys SB RAS, Krasnoyarsk 660036, Russia.
Siberian Fed Univ, Krasnoyarsk 660041, Russia.

Доп.точки доступа:
Bashmakova, Eugenia E.; Panamarev, Nikita S.; Kudryavtsev, Alexander N.; Frank, Ludmila A.; Kudryavtsev, Alexander; Russian Federation for young scientists [MK-772.2020.4]; Government of Krasnoyarsk Territory, Krasnoyarsk Regional Science Foundation [2021012006966]

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