己糖激酶2在犬乳腺肿瘤中的表达及预后研究

doi:10.11843/j.issn.0366-6964.2023.03.041

摘要/Abstract

摘要： 本研究旨在探究己糖激酶2(hexokinase 2, HK2)在犬乳腺肿瘤(canine mammary tumor, CMT)中的表达情况及与患犬预后的相关性。将临床收集的121例犬乳腺肿块样本制作石蜡切片，经组织病理学分析判读后随机抽取恶性乳腺肿瘤与正常乳腺组织样本各3份进行高通量转录组测序，筛选差异表达基因。使用实时荧光定量PCR(quantitative real time PCR，qPCR)和免疫组织化学法检测HK2在犬全身组织及乳腺肿瘤组织中的表达情况，对术犬长期随访调查并分析HK2与乳腺肿瘤患犬临床特征及预后的相关性。结果表明：1)本研究中收集到的121例犬乳腺肿块临床样本中良性乳腺肿瘤33例，恶性乳腺肿瘤78例；2)经高通量转录组测序筛选出HK2、MEST、GDF5等36个显著差异表达基因，其中,HK2经qPCR验证以及基于其在细胞代谢中的关键作用且在人类某些肿瘤中异常表达，故将HK2作为研究对象；3)HK2 mRNA与蛋白表达水平在犬正常乳腺组织中均表现为低表达，而CMT中HK2 mRNA与蛋白表达水平均显著升高(P＜0.01)；4)同时HK2的表达水平与肿瘤组织类型、肿瘤大小、脉管浸润、Ki-67指数以及预后显著相关(P＜0.01)。HK2在CMT组织中呈高表达，并与肿瘤的临床特征及预后显著相关。HK2具有作为CMT的诊断与预后评估因子的潜力。

关键词: 犬乳腺肿瘤, 己糖激酶2, 糖代谢, 高通量测序

Abstract: This experiment was conducted to study the expression of hexokinase 2 (HK2) in canine mammary tumor (CMT) and its correlation with the prognosis of dogs. Canine mammary masses from 121 clinical samples were made into paraffin sections. After histopathological analysis and interpretation, 3 samples of malignant mammary tumors and 3 samples of normal mammary gland tissues were randomly selected for high-throughput transcriptome sequencing to screen differentially expressed genes. Quantitative real time PCR (qPCR) and immunohistochemistry were applied to detect the expression of HK2 in the overall body tissues and mammary tumors, and long-term follow-up of dogs was conducted to investigate and analyze the correlation between HK2 and the clinical characteristics and prognosis of dogs with mammary tumor. The results showed that: 1) There were 33 benign and 78 malignant mammary tumors in 121 clinical samples of canine mammary masses collected in this study; 2) Thirty-six significantly and differentially expressed genes such as HK2, MEST, GDF5 and etc, were screened out by high-throughput transcriptome sequencing. Based on the verification by qPCR and the key role played in cell metabolism, and abnormal expression in some human tumors. HK2 was designated as the research object; 3) The expression levels of HK2 mRNA and protein were low in normal canine mammary gland tissues, but significantly elevated in CMT (P<0.01); 4) The expression level of HK2 was significantly correlated with tumor tissue type, tumor size, vascular invasion, Ki-67 index and prognosis (P<0.01). HK2 is highly expressed in CMT tissues, and is significantly related to the clinical characteristics and prognosis of CMT. HK2 can be potentially regarded as a diagnostic and prognostic factor of CMT.

Key words: canine mammary tumor, hexokinase 2, glucose metabolism, high throughput sequencing

中图分类号:

胥辉豪, 刘江渝, 李启卷, 郑小波, 林珈好, 金艺鹏, 林德贵. 己糖激酶2在犬乳腺肿瘤中的表达及预后研究[J]. 畜牧兽医学报, 2023, 54(3): 1310-1324.

XU Huihao, LIU Jiangyu, LI Qijuan, ZHENG Xiaobo, LIN Jiahao, JIN Yipeng, LIN Degui. Expression and Prognosis of Hexokinase 2 in Canine Mammary Tumors[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1310-1324.

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