摘要
目的目前乳腺癌发病率已居女性恶性肿瘤之首,因此需要潜在的生物标记物来预测及预防乳腺癌。文中旨在通过建立乳腺癌组织miRNA表达谱,探讨差异miRNA及其靶基因表达在乳腺癌中的表达及临床意义。方法收集2015年1月至2018年12月贵州医科大学附属医院乳腺外科137例手术切除的乳腺癌组织及癌旁组织及20例乳腺纤维腺瘤组织。将标本组织分为乳腺癌组、癌旁组、乳腺纤维腺瘤组,淋巴结转移组。利用高通量测序技术检测乳腺癌组织及其配对癌旁组织的miRNA,Real-time PCR验证表达差异最为显著的3个miRNA在不同组中表达情况,采用生物信息学分析技术预测其靶基因,且观察靶基因的miRNA及蛋白在不同乳腺疾病中的表达情况。结果高通量测序共筛选出157个表达上调和162个表达下调的miRNA,其中上调最显著为hsa-miR-532-3p和hsa-miR-1260b,下调最显著为hsa-let-7c-5p,生物信息学分析其靶基因均为EZH2。与癌旁组比较,乳腺癌组、淋巴结转移组hsa-miR-532-3p、hsa-miR-1260b表达明显上调(P<0.05);乳腺癌组、淋巴结转移组hsa-let-7c-5p明显下调(P<0.05)。乳腺纤维腺瘤组、乳腺癌组、淋巴结转移组EZH2 miRNA表达量(1.24±0.01,4.02±0.01,15.97±0.01)较癌旁组(1.00±0.00)明显升高(P<0.05);EZH2 蛋白表达量亦明显升高(P<0.05)。结论 hsa-miR-532-3p、hsa-miR-1260b及hsa-let-7c-5p与乳腺癌的发生密切相关,其可能通过诱导EZH2的转录表达,进而促进乳腺癌发生发;hsa-miR-532-3p、hsa-miR-1260b、hsa-let-7c-5p及EZH2可能成为乳腺癌潜在的肿瘤生物标志物。
Objective Recently, the incidence of breast cancer has been the highest among female malignant tumors. Therefore, potential biomarkers are urgently needed to predict and prevent breast cancer. This study was aimed to explore the expression and clinical significance of differential miRNAs and their target genes in breast cancer by establishing miRNA expression profile in breast cancer tissues. Methods From January 2015 to December 2018, a total of 137 cases of breast cancer tissues with paired paracancerous tissues and 20 cases of breast fibroadenoma tissues were collected from the department of breast surgery, affiliated hospital of Guizhou Medical University. The tissues were divided into breast cancer group, paracancerous group, breast fibroadenoma group and lymph node metastasis group. High-throughput sequencing technique was used to detect the miRNAs in breast cancer tissues and paired paracancerous tissues. Real-time PCR verified the expressions of the three miRNAs with the most significant expression differences in different groups. Finally, bioinformatics analysis was used to predict the target genes and investigate the expression of miRNAs and proteins of target genes in different breast diseases. Results A total of 157 upregulated and 162 downregulated miRNAs were screened by high-throughput sequencing. Mir-hsa-miR-532-3p and hsa-miR-1260b were the most significant in upregulated miRNAs while has-let-7c-5p was the most significant in downregulated miRNAs. Meanwhile, bioinformatics analysis showed that their target gene was EZH2. Compared with para-cancerous group, expressions of hsa-miR-532-3p and hsa-miR-1260b were significantly upregulated while hsa-let-7c-5p was significantly down-regulated in breast cancer group and lymph node metastasis group (all P<0.05). The miRNA expressions of target gene EZH2 in breast cancer group, breast fibroadenoma group and lymph node metastasis group (1.24±0.01, 4.02±0.01, 15.97±0.01, respectively) were upregulated when compared with the para-cancerous group (1.00±0.00), and the similar conclusion could be drawn in EZH2 protein expression. Conclusion Hsa-miR-532-3p, hsa-miR-1260b and hsa-let-7c-5p were closely related to breast cancer, which may promote the occurrence and development of breast cancer by inducing the transcriptional expression of EZH2. HsamiR-532-3p, hsa-miR-1260b, hsa-let-7c-5p and EZH2 may be potential tumor biomarkers of breast cancer.
作者
李朝夕
黄健
张姝
何芸
陆景润
刘敏
莫非
罗扬勇
刘晋廷
黄建军
LI Zhao-xi;HUANG Jian;ZHANG Shu;HE Yun;LU Jing-run;LIU Min;MO Fei;LUO Yang-yong;LIU Jin-ting;HUANG Jian-jun(Guizhou Medical University,Guiyang 550001,Guizhou,China;Clinical Laboratory Teaching and Researching Department,Army Medical University,Chongqing 400038,China;Clinical Laboratory Center,Affiliated Hospital of Guizhou Medical University,Guiyang 550001,Guizhou,China;Department of Breast Surgery, Affiliated Hospital of Guizhou Medical University,Guiyang 550001,Guizhou,China)
出处
《医学研究生学报》
CAS
北大核心
2019年第8期833-839,共7页
Journal of Medical Postgraduates
基金
中国博士后科学基金(2018M643862)
贵州省科技厅科学技术基金(黔科合J字[2013]2047号)
