A.A. Osipova1, D.A. Ryabchikov2, S.V. Chulkova3, A.M. Burdennyy4, V.I. Loginov5, N.E. Kushlinskii6

1,6 Russian University of Medicine (Moscow, Russia)

2,3,6 N.N. Blokhin National Medical Research Center of Oncology of the Ministry of Health of Russia (Moscow, Russia)

3 I.I. Pirogov Russian National Research Medical University of the Russian Ministry of Health (Moscow, Russia)

4,5 Research Institute of General Pathology and Pathophysiology (Moscow, Russia)

5 Research Centre for Medical Genetics, (Moscow, Russia)

1 osipova_ronc@mail.ru

lncRNAs are transcripts that play a regulatory role in many biological processes, and their expression profile is associated with cell proliferation, migration, and invasion. Methylation of lncRNA genes can lead to disruption of their functions.

The purpose of the work is to study the methylation status of the lncRNA genes MEG3, SEMA3B-AS1 in luminal breast cancer and assess their clinical significance.

The study included 367 breast cancer patients. The predominant stage was T1 – 142 (38.7%) and T2 – 204 (55.6%) stages, N0 and N1 status, respectively, 198 (54.0%) and 139 (37.9%). Luminal A was detected in 28.6%, B Her2neu“-” – in 30.3%, B Her2neu“+” – in 27.2%, Erb-B2, an overexpressing subtype, was detected in 6.3% and 7.6 % triple-negative subtype. In 77.1% of cases - invasive ductal, in 12.8% - lobular, mainly G2 - 62.9%. We studied the methylation of CpG regions of the promoter regions of the lncRNA genes MEG3 and SEMA3B-AS1 in paired images of tumor and histologically unchanged mammary gland tissue using methyl-specific polymerase chain reaction (MS-PCR). Amplifier T100 (“Bio-Rad”, USA). Primers were selected using the Lasergene 17.1 software package from DNASTAR (USA). Statistical data processing was carried out using SPSS, v. 23. It was found that the frequency of methylation of the MEG3 and SEMA3B-AS1 genes in tumor tissue was significantly higher than in histologically unchanged tissue. Methylation of the studied genes is associated with age (R = -0.24 for MEG3 and R = -0.36 for SEMA3B-AS1) and increases with increasing stage of the tumor process. A correlation was established between methylation of the MEG3 and SEMA3B-AS1 genes with tumor stage (R=0.46 and R=0.33, respectively, p<0.05), T index (R=0.43 and R=0.36, respectively, p<0.05). As the tumor stage increases (I – II – III), the frequency of gene methylation increases. In tumor tissue, MEG3 and SEMA3B-AS1 were comethylated (R=0.79, p<0.05). Methylation of MEG3, SEMA3B-AS1 is associated with a decrease in 5- and 10-year overall survival (OS) and progression-free time (PFS) in luminal breast cancer stages I-II.

In case of breast cancer in tumor tissue, the frequency of methylation of lncRNA genes MEG3, SEMA3B-AS1 exceeds that in histologically unchanged tissue, which confirms the suppressor activity of these genes. The relationship between the stage, size of the primary tumor, OS, and PFS with the high frequency of methylation of the MEG3 and SEMA3B-AS1 genes in tumor tissue allows us to consider them as prognostic markers.

Osipova A.A., Ryabchikov D.A., Chulkova S.V., Burdennyy A.M., Loginov V.I., Kushlinskii N.E. Clinical significance of methylation of lncRNA genes MEG3, SEMA3B-AS1 in tumors of patients with luminal breast cancer. Technologies of Living Systems. 2024. V. 21. № 3. Р. 74-83. DOI: https://doi.org/10.18127/j20700997-202403-08 (In Russian).

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