danДоклады Национальной академии наук БеларусиDoklady of the National Academy of Sciences of Belarus1561-83232524-2431The Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1561-8323-2023-67-4-300-306dan-1141Research ArticleБИОЛОГИЯBIOLOGYОпределение статуса метилирования промоторных областей генов MARCH11, HOXA9, PTGDR и UNCX у пациентов с немелкоклеточным раком легкогоDetermining the methylation status of the promoter regions of MARCH11, HOXA9, PTGDR, and UNCX genes in patients with non-small cell lung cancerМихаленкоЕ. П.MikhalenkaA. P.

Михаленко Елена Петровна – кандидат биологических наук, старший научный сотрудник

ул. Академическая, 27, 220072, Минск

Mikhalenka Alena P. – Ph. D. (Biology), Leading Researcher

27, Akademicheskaya Str., 220072, Minsk

E.Michalenko@igc.byЩаюкА. Н.ShchayukA. N.

Щаюк Анна Николаевна – кандидат биологических наук, старший научный сотрудник

ул. Академическая, 27, 220072, Минск

Shchayuk Anna N. – Ph. D. (Biology), Senior Researcher

27, Akademicheskaya Str., 220072, Minsk

anna.shchayuk@tut.byНикитинскаяТ. В.NikitinskayaT. V.

Никитинская Татьяна Владимировна – научный сотрудник

ул. Академическая, 27, 220072, Минск

Nikitinskaya Tatsiana V. – Researcher. Institute of Genetics and Cytology

27, Akademicheskaya Str., 220072, Minsk

T.Nikitinskaya@igc.byПолюховичЮ. В.PaliukhovichYu. V.

Полюхович Юлия Владимировна – кандидат биологических наук, старший научный сотрудник

ул. Академическая, 27, 220072, Минск

Paliukhovich Yuliya V. – Ph. D. (Biology), Senior Researcher

27, Akademicheskaya Str., 220072, Minsk

yu.paliukhovich@igc.byКубракС. В.KubrakS. V.

Кубрак Светлана Владимировна – кандидат биологических наук, старший научный сотрудник

ул. Академическая, 27, 220072, Минск

Kubrak Sviatlana V. – Ph. D. (Biology), Leading Researcher

27, Akademicheskaya Str., 220072, Minsk

S.Kubrak@igc.byШепетькоМ. Н.ShapetskaM. N.

Шепетько Михаил Николаевич – кандидат медицинских наук, доцент

пр. Дзержинского, 83, 220116, Минск

Shapetska Michail N. – Ph. D. (Medicine), Assistant Professor

83, Dzerzhinsky Ave., 220116, Minsk

shepetjko@gmail.comКильчевскийА. В.KilchevskyA. V.

Кильчевский Александр Владимирович – академик, доктор биологических наук, профессор, главный научный сотрудник

ул. Академическая, 27, 220072, Минск

Kilchevsky Aleksandr V. – Academician, D. Sc. (Biology), Professor, Chief Researcher

27, Akademicheskaya Str., 220072, Minsk

kilchev@presidium.bas-net.byИнститут генетики и цитологии Национальной академии наук БеларусиInstitute of Genetics and Cytology of the National Academy of Sciences of BelarusБелорусский медицинский университетBelarusian Medical University202301092023674300306Copyright © Михаленко Е.П., Щаюк А.Н., Никитинская Т.В., Полюхович Ю.В., Кубрак С.В., Шепетько М.Н., Кильчевский А.В., 20232023Михаленко Е.П., Щаюк А.Н., Никитинская Т.В., Полюхович Ю.В., Кубрак С.В., Шепетько М.Н., Кильчевский А.В.Mikhalenka A.P., Shchayuk A.N., Nikitinskaya T.V., Paliukhovich Y.V., Kubrak S.V., Shapetska M.N., Kilchevsky A.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://doklady.belnauka.by/jour/article/view/1141

Целью данного исследования было определение статуса метилирования промоторных областей генов MARCH11, HOXA9, PTGDR и UNCX в опухолевой и неопухолевой ткани легкого у пациентов с немелкоклеточным раком легкого (НМРЛ). Относительный уровень метилирования промоторных областей генов MARCH11, HOXA9, PTGDR, UNCX определяли методом количественной метилспецифической ПЦР у 73 пациентов с НМРЛ. Количественная метилспецифическая реакция проводилась как для образцов опухолевой ткани, так и для образцов неопухолевой ткани одного и того же пациента. Для каждого из образцов ставили реакцию по изучаемым генам (MARCH11, UNCX, HOXA9, PTGDR) и по референсному гену бета-актина (β-actin). Установлены положительные уровни метилирования гена HOXA9 у 83,5 % пациентов, гена MARCH11 – у 80,8 % пациентов, гена PTGDR – у 68,4 % пациентов, гена UNCX – у 84,9 % пациентов. В исследуемой группе пациентов с НМРЛ выявлены достоверные различия относительных уровней метилирования промоторных регионов генов MARCH11, HOXA9, PTGDR и UNCX в опухолевой и неопухолевой ткани легкого. Эти данные позволяют предположить, что гиперметилирование генов MARCH11, HOXA9, PTGDR и UNCX может играть роль в опухолевой прогрессии НМРЛ.

The aim of this study was to determine the methylation status of the promoter regions of MARCH11, HOXA9, PTGDR, and UNCX genes in the tumor and non-tumor lung tissue in patients with non-small cell lung cancer (NSCLC). A relative level of methylation of the promoter regions of MARCH11, HOXA9, PTGDR, and UNCX genes was determined by the quantitative methylation-specific PCR in 73 patients with NSCLC. The quantitative methylation-specific reaction was performed both for tumor tissue samples and non-tumor tissue samples of the same patient. For each of the samples, a reaction was set both by the investigated genes (MARCH11, UNCX, HOXA9, and PTGDR) and by the reference beta-actin gene (β-actin). Positive levels of methylation of the HOXA9 gene were established for 83.5 % patients; the MARCH11 gene – for 80.8 % patients; the PTGDR gene – for 68.4 % patients; the UNCX gene – for 84.9 % patients. In the study group of patients with NSCLC, significant differences were found in the relative levels of methylation of the promoter regions of MARCH11, HOXA9, PTGDR, and UNCX genes in the tumor and non-tumor lung tissue. The data suggest that hypermethylation of MARCH11, HOXA9, PTGDR, and UNCX genes may play a role in NSCLC tumor progression.

эпигенетикаколичественная метилспецифическая реакцияотносительный уровень метилированиянемелкоклеточный рак легкогоопухолевая и неопухолевая тканьepigeneticsquantitative methyl-specific reactionrelative methylation levelnon-small cell lung cancertumor and non-tumor tissueReferencesZhang, L. Epigenetics in health and disease / L. Zhang, Q. Lu, C. Chang // Adv. Exp. Med. Biol. – 2020. – Vol. 1253. – P. 3–55. https://doi.org/10.1007/978-981-15-3449-2_1Zhang L., Lu Q., Chang C. Epigenetics in health and disease. Advances in Experimental Medicine and Biology, 2020, vol. 1253, pp. 3–55. https://doi.org/10.1007/978-981-15-3449-2_1Bump hunting to identify differentially methylated regions in epigenetic epidemiology studies / A. E. Jaffe [et al.] // Int. J. Epidemiol. – 2012. – Vol. 41, N 1. – P. 200–209. https://doi.org/10.1093/ije/dyr238Jaffe A. E., Murakami P., Lee H., Leek J. T., Fallin M. D., Feinberg A. P., Irizarry R. A. Bump hunting to identify differentially methylated regions in epigenetic epidemiology studies. International Journal of Epidemiology, 2012, vol. 41, no. 1, pp. 200–209. https://doi.org/10.1093/ije/dyr238A novel epigenetic signature for early diagnosis in lung cancer / A. Diaz-Lagares [et al.] // Clin. Cancer Res. – 2016. – Vol. 22, N 13. – P. 3361–3371. https://doi.org/10.1158/1078-0432.ccr-15-2346Diaz-Lagares A, Mendez-Gonzalez J., Hervas D., Saigi M., Pajares M. J., Garcia D., Crujerias A. B., Pio R., Montuenga L. M., Zulueta J., Nadal E., Rosell A., Esteller M., Sandoval J. A novel epigenetic signature for early diagnosis in lung cancer. Clinical Cancer Research, 2016, vol. 22, no. 13, pp. 3361–3371. https://doi.org/10.1158/1078-0432.ccr-15-2346Pan-cancer predictions of transcription factors mediating aberrant DNA methylation / D. Detilleux [et al.] // Epigenetics and Chromatin. – 2022. – Vol. 15, N 1. https://doi.org/10.1186/s13072-022-00443-wDetilleux D., Spill Y. G., Balaramane D., Weber M., Bardet A. F. Pan-cancer predictions of transcription factors mediating aberrant DNA methyl ation. Epigenetics and Chromatin, 2022, vol. 15, no. 1. https://doi.org/10.1186/s13072-022-00443-wFukushige, S. DNA methylation in cancer: a gene silencing mechanism and the clinical potential of its biomarkers / S. Fukushige, A. Horii // Tohoku J. Exp. Med. – 2013. – Vol. 229, N 3. – P. 173–185. https://doi.org/10.1620/tjem.229.173Fukushige S., Horii A. DNA methylation in cancer: a gene silencing mechanism and the clinical potential of its biomarkers. Tohoku Journal of Experimental Medicine, 2013, vol. 229, no. 3, pp. 173–185. https://doi.org/10.1620/tjem.229.173Differentially methylated regions within lung cancer risk loci are enriched in deregulated enhancers / M. Laplana [et al.] // Epigenetics. – 2021. – Vol. 17, N 2. – P. 117–132. https://doi.org/10.1080/15592294.2021.1878723Laplana M., Bieg M., Faltus C., Melnik S., Bogatyrova O., Gu Z., Muley T., Meister M., Dienemann H., Herpel E., Amos C. I., Schlesner M., Eils R., Plass C., Risch A. Differentially methylated regions within lung cancer risk loci are enriched in deregulated enhancers. Epigenetics, 2021, vol. 17, no. 2, pp. 117–132. https://doi.org/10.1080/15592294.2021.1878723Methylation profiling defines an extensive field defect in histologically normal prostate tissues associated with prostate cancer / B. Yang [et al.] // Neoplasia. – 2013. – Vol. 15, N 4. – P. 399–408. https://doi.org/10.1593/neo.13280Yang B., Bhusari S., Kueck J., Weeratunga P., Wagner J., Leverson G., Huang W., Jarrard D. F. Methylation profiling defines an extensive field defect in histologically normal prostate tissues associated with prostate cancer. Neoplasia, 2013, vol. 15, no. 4, pp. 399–408. https://doi.org/10.1593/neo.13280A Panel of Novel Detection and Prognostic Methylated DNA Markers in Primary Non–Small Cell Lung Cancer and Serum DNA / A. Ooki [et al.] // Clin. Cancer Res. – 2017. – Vol. 23, N 22. – P. 7141–7152. https://doi.org/10.1158/1078-0432.ccr-17-1222Ooki A., Maleki Z., Tsay J.-C. J., Goparaju Ch., Brait M., Turaga N., Nam H.-S., Rom W. N., Pass H. I., Sidransky D., Guerrero-Preston R., Hoque M. O. A Panel of Novel Detection and Prognostic Methylated DNA Markers in Primary Non– Small Cell Lung Cancer and Serum DNA. Clinical Cancer Research, 2017, vol. 23, no. 22, pp. 7141–7152. https://doi.org/10.1158/1078-0432.ccr-17-1222Epigenome-wide scan identifies differentially methylated regions for lung cancer using pre-diagnostic peripheral blood / Naisi Zhao [et al.] // Epigenetics. – 2022. – Vol. 17, N 4. – P. 460–472. https://doi.org/10.1080/15592294.2021.1923615Zhao N., Ruan M., Koestler D. C., Lu J., Marsit C. J., Kelsey K. T., Platz E. A., Michaud D. S. Epigenome-wide scan identifies differentially methylated regions for lung cancer using pre-diagnostic peripheral blood. Epigenetics, 2022, vol. 17, no. 4, pp. 460–472. https://doi.org/10.1080/15592294.2021.1923615Methylation-associated inactivation of JPH3 and its effect on prognosis and cell biological function in HCC / Yi Huang [et al.] // Mol. Med. Rep. – 2022. – Vol. 25, N 4. – Art. 124. https://doi.org/10.3892/mmr.2022.12640Huang Yi, Yu Z., Zheng M., Yang X., Huang H., Zhao L. Methylation-associated inactivation of JPH3 and its effect on prognosis and cell biological function in HCC. Molecular Medicine Reports, 2022, vol. 25, no. 4, art. 124. https://doi.org/10.3892/mmr.2022.12640DNA hypermethylation and decreased mRNA expression of MAL, PRIMA1, PTGDR and SFRP1 in colorectal adenoma and cancer / A. Kalmar [et al.] // BMC Cancer. – 2015. – Vol. 15, N 1. – Art. 736. https://doi.org/10.1186/s12885-015-1687-xKalmár A., Péterfia B., Hollósi P., Galamb O., Spisák S., Wichmann B., Bodor A., Tóth K., Patai Á. V., Valcz G., Nagy Z. B., Kubák V., Tulassay Z., Kovalszky I., Molnár B. DNA hypermethylation and decreased mRNA expression of MAL, PRIMA1, PTGDR and SFRP1 in colorectal adenoma and cancer. BioMed Central Cancer, 2015, vol. 15, no. 1, art. 736. https://doi.org/10.1186/s12885-015-1687-xPradhan, M. P. Systems biology approach to stage-wise characterization of epigenetic genes in lung adenocarcinoma / M. P. Pradhan, A. Desai, M. J. Palakal // BMC Syst. Biol. – 2013. – Vol. 7, N 1. – Art. 141. https://doi.org/10.1186/1752-0509-7-141Pradhan M. P., Desai A., Palakal M. J. Systems biology approach to stage-wise characterization of epigenetic genes in lung adenocarcinoma. BMC Systems Biology, 2013, vol. 7, no. 1, art. 141. https://doi.org/10.1186/1752-0509-7-141Epigenetically induced ectopic expression of UNCX impairs the proliferation and differentiation of myeloid cells / G. Daniele [et al.] // Haematologica. – 2017. – Vol. 102, N 7. – P. 1204–1214. https://doi.org/10.3324/haematol.2016.163022Daniele G., Simonetti G., Fusilli C., Iacobucci I., Lonoce A., Palazzo A., Lomiento M. [et al.]. Epigenetically induced ectopic expression of UNCX impairs the proliferation and differentiation of myeloid cells. Haematologica, 2017, vol. 102, no. 7, pp. 1204–1214. https://doi.org/10.3324/haematol.2016.163022The prognostic value of homeobox A9 (HOXA9) methylation in solid tumors: a systematic review and meta-analysis / H. Cai [et al.] // Transl. Cancer Res. – 2021. – Vol. 10, N 10. – P. 4347–4354. https://doi.org/10.21037/tcr-21-765Cai H., Ke Z. B., Dong R. N., Chen H., Lin F., Zheng W. C., Chen S. H., Zhu J. M., Chen S. M., Zheng Q. S., Wei Y., Xue X. Y., Xu N. The prognostic value of homeobox A9 (HOXA9) methylation in solid tumors: a systematic review and metaanalysis. Translation Cancer Research, 2021, vol. 10, no. 10, pp. 4347–4354. https://doi.org/10.21037/tcr-21-765Methylation of HOXA9 and ISL1 Predicts Patient Outcome in High-Grade Non-Invasive Bladder Cancer / M. O. Kitchen [et al.] // PLoS One. – 2015. – Vol. 10, N 9. – Art. e0137003. https://doi.org/10.1371/journal.pone.0137003Kitchen M. O., Bryan R. T., Haworth K. E., Emes R. D., Luscombe Ch., Gommersall L., Cheng K. K., Zeegers M. P., James N. D., Devall A. J., Fryer A. A., Farrell W. E. Methylation of HOXA9 and ISL1 Predicts Patient Outcome in HighGrade Non-Invasive Bladder Cancer. PLoS One, 2015, vol. 10, no. 9, art. e0137003. https://doi.org/10.1371/journal.pone.0137003

The authors declare that there are no conflicts of interest present.