Szewczuk Wiktor, Szewczuk Oksana, Czajkowski Krzysztof, Gromadka Robert, Man Yan-Gao, Wałędziak Maciej, Semczuk Andrzej
Department of Pathology, Military Institute of Medicine, Warsaw, Poland.
Department of Gynecology and Gynecologic Oncology, Military Institute of Medicine, Warsaw, Poland.
J Cancer. 2024 May 30;15(13):4040-4046. doi: 10.7150/jca.94945. eCollection 2024.
Advanced-stage ovarian cancer (OC) is among the most fatal female genital tract neoplasms worldwide. Although different genetic mechanisms have been shown to be involved in ovarian carcinogenesis, the role of introns methylation is still unresolved. We performed methylation analysis of introns 1, 3, and 4 of the to identify patterns in primary stage III OCs, corresponding metastases, and healthy tissues. The study involved samples of paraffin-embedded tissues obtained from 80 patients with stage III OCs, who underwent surgery at the Department of Gynecology and Gynecologic Oncology of the Military Institute of Medicine in Warsaw, Poland. Altogether, 40 serous-type G2/3 OCs and 40 endometrioid-type G2/3 OCs were included. From the same patient, metastatic and normal tissues were simultaneously analyzed. As a control group, 80 tissue samples were collected from patients after bariatric operations. Human ovarian cancer A2780 cell line was also investigated. Total genomic DNA was isolated from paraffin-embedded tissue blocks and the methylation analysis was performed by bisulfite DNA conversion, DNA amplification with specific primers, cloning, and DNA sequencing. All of the samples of intron 1 of were un-methylated in OCs, metastatic tissues, and in healthy tissues from the same patient. Also, no methylation of intron 1 was detected in cells from the human A2780 ovarian cancer cell line and in all samples from control group. In all samples, introns 3 and 4 of the were methylated in primary tumors, metastatic tissue, and in healthy tissue from the same patient, in human A2780 ovarian cell line, and in DNA samples from healthy patients. None of the clinicopatholocal features was related to the introns methylation status. Our data on introns methylation sheds new light on the mechanism of p53 activity for a better understanding of cancer biology. The study suggests the existence of an additional regulation rule of activity that involves demethylation-methylation mechanisms. Methylation at introns 3 and 4 may also overall help in protecting against damage by viral restrictases or viral DNA integration.
晚期卵巢癌（OC）是全球最致命的女性生殖道肿瘤之一。尽管已显示不同的遗传机制参与卵巢癌的发生，但内含子甲基化的作用仍未明确。我们对[具体基因名称]的内含子1、3和4进行甲基化分析，以确定原发性III期OC、相应转移灶和健康组织中的模式。该研究涉及从80例III期OC患者获取的石蜡包埋组织样本，这些患者在波兰华沙军事医学研究所妇产科和妇科肿瘤学系接受了手术。总共纳入了40例浆液性G2/3 OC和40例子宫内膜样G2/3 OC。从同一患者同时分析转移组织和正常组织。作为对照组，从接受减肥手术的患者中收集了80个组织样本。还对人卵巢癌A2780细胞系进行了研究。从石蜡包埋组织块中分离出总基因组DNA，并通过亚硫酸氢盐DNA转化、用特异性引物进行DNA扩增、克隆和DNA测序进行甲基化分析。[具体基因名称]内含子1的所有样本在OC、转移组织以及同一患者的健康组织中均未甲基化。此外，在人A2780卵巢癌细胞系的细胞以及对照组的所有样本中均未检测到[具体基因名称]内含子1的甲基化。在所有样本中，[具体基因名称]的内含子3和4在原发性肿瘤、转移组织、同一患者的健康组织、人A2780卵巢细胞系以及健康患者的DNA样本中均发生甲基化。没有任何临床病理特征与[具体基因名称]内含子甲基化状态相关。我们关于[具体基因名称]内含子甲基化的数据为p53活性机制提供了新的线索，以更好地理解癌症生物学。该研究表明存在一种涉及去甲基化 - 甲基化机制的[具体基因名称]活性的额外调控规则。内含子3和4处的甲基化也可能总体上有助于保护[具体基因名称]免受病毒限制酶或病毒DNA整合的损伤。
