鉴定和验证 DNA 甲基化标志物，以预测乳腺癌的腋窝淋巴结转移。
Identification and validation of DNA methylation markers to predict axillary lymph node metastasis of breast cancer.
机构信息
Department of Breast Surgery, Shenzhen Maternity & Child Healthcare Hospital of Southern Medical University, Shenzhen, Guangdong, China.
出版信息
PLoS One. 2022 Dec 1;17(12):e0278270. doi: 10.1371/journal.pone.0278270. eCollection 2022.
BACKGROUND
Axillary lymph node metastasis (ALNM) is one of the most important prognostic factors for breast cancer patients, and DNA methylation is involved in ALNM of breast cancer. However, the methylation profile of breast cancer ALNM remains unknown.
METHODS
Breast cancer tissues were collected from patients with and without ALNM. We investigated the genome-wide DNA methylation profile in breast cancer with and without ALNM using reduced representation bisulfite sequencing (RRBS). Then, differentially methylated regions (DMRs) were verified by targeted bisulfite sequencing.
RESULTS
A total of 21491 DMRs were identified between the lymph node positive group and negative group. Compared to the LN-negative breast cancer, LN-positive breast cancer had 10,920 hypermethylated DMRs and 10,571 hypomethylated DMRs. Then, 10 DMRs in the gene promoter region were detected by targeted bisulfite sequencing, these gene included HOXA5, PTOV1-AS1, RHOF, PAX6, GSTP1, RASGRF2, AKR1B1, BNIP3, CRMP1, ING5. Compared with negative lymph node, the promoter methylation levels of RASGRF2, AKR1B1 and CRMP1 increased in positive lymph node, while the promoter methylation level of RHOF decreased in positive lymph node. In addition, Cancer Genome Atlas (TCGA) data showed that RASGRF2, AKR1B1 and CRMP1 were low expressed in breast Cancer tissues, while RHOF was high expressed in breast Cancer tissues. Furthermore, in addition to highly methylated AKR1B1, RASGRF2 and CRMP1 gene promoters, BNIP3, GSTP1, HOXA5 and PAX6 gene promoters were also methylated in ER-positive and HER2-negative breast cancer with ALNM.
CONCLUSIONS
When compared to negative lymph node breast cancer, the positive lymph node breast cancer has a differential methylation status. Promoter methylation of RASGRF2, AKR1B1, CRMP1 and RHOF in lymph node positive breast cancer tissues was significantly different from that in lymph node negative breast cancer tissues. AKR1B1, RASGRF2, CRMP1, BNIP3, GSTP1, HOXA5 and PAX6 genes were methylated in ER-positive and HER2-negative breast cancer with ALNM. The study provides an important biological base for understanding breast cancer with ALNM and developing therapeutic targets for breast cancer with ALNM.
背景
腋窝淋巴结转移（ALNM）是乳腺癌患者最重要的预后因素之一，而 DNA 甲基化参与了乳腺癌的 ALNM。然而，乳腺癌 ALNM 的甲基化谱尚不清楚。
方法
收集有和无 ALNM 的乳腺癌患者的组织。我们使用简化重亚硫酸盐测序（RRBS）检测有和无 ALNM 的乳腺癌的全基因组 DNA 甲基化谱。然后，通过靶向重亚硫酸盐测序验证差异甲基化区域（DMRs）。
结果
在淋巴结阳性组和阴性组之间共鉴定出 21491 个 DMR。与 LN-阴性乳腺癌相比，LN-阳性乳腺癌有 10920 个高甲基化 DMR 和 10571 个低甲基化 DMR。然后，通过靶向重亚硫酸盐测序检测到 10 个基因启动子区域的 DMR，这些基因包括 HOXA5、PTOV1-AS1、RHOF、PAX6、GSTP1、RASGRF2、AKR1B1、BNIP3、CRMP1 和 ING5。与阴性淋巴结相比，RASGRF2、AKR1B1 和 CRMP1 的启动子甲基化水平在阳性淋巴结中升高，而 RHOF 的启动子甲基化水平在阳性淋巴结中降低。此外，癌症基因组图谱（TCGA）数据显示，RASGRF2、AKR1B1 和 CRMP1 在乳腺癌组织中低表达，而 RHOF 在乳腺癌组织中高表达。此外，除了高甲基化的 AKR1B1、RASGRF2 和 CRMP1 基因启动子外，ER 阳性和 HER2 阴性伴 ALNM 的乳腺癌中 BNIP3、GSTP1、HOXA5 和 PAX6 基因启动子也发生了甲基化。
结论
与阴性淋巴结乳腺癌相比，阳性淋巴结乳腺癌存在差异甲基化状态。与阴性淋巴结乳腺癌相比，阳性淋巴结乳腺癌组织中 RASGRF2、AKR1B1、CRMP1 和 RHOF 的启动子甲基化差异显著。AKR1B1、RASGRF2、CRMP1、BNIP3、GSTP1、HOXA5 和 PAX6 基因在 ER 阳性和 HER2 阴性伴 ALNM 的乳腺癌中发生甲基化。该研究为理解伴有 ALNM 的乳腺癌提供了重要的生物学基础，并为伴有 ALNM 的乳腺癌治疗靶点的开发提供了依据。
Zotero 插件