HER2阳性乳腺癌中p95HER2表达与细胞增殖的相关性研究  

Correlation between p95HER2 expression and cell proliferation in HER2-positive breast cancer

作　　者：吴旸 张薇[2] 董丽平 徐迪[2] 邓飞 唐金海[2] WU Yang;ZHANG Wei;DONG Liping;XU Di;DENG Fei;TANG Jinhai(Biobank,the First Af liated Hospital of Nanjing Medical University,Nanjing 210029,China;Department of General Surgery,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China;Department of General Surgery,Nanjing Pukou People’s Hospital,Nanjing 210029,China)

机构地区：[1]南京医科大学第一附属医院生物样本库,江苏南京210029 [2]南京医科大学第一附属医院普外科,江苏南京210029 [3]南京市浦口人民医院普外科,江苏南京210029

出　　处：《中国肿瘤外科杂志》2023年第5期423-430,443,共9页Chinese Journal of Surgical Oncology

基　　金：国家自然科学基金青年项目(81901883)。

摘　　要：目的通过检测并分析HER2阳性乳腺癌患者肿瘤组织中p95HER2的表达情况,探索其与相关临床及病理特征之间的关系,建立p95HER2过表达细胞验证其促进细胞增殖的生物学功能,为临床HER2阳性乳腺治疗提供实验基础。方法收集2020年12月至2021年12月就诊于江苏省人民医院普外科的47例HER2阳性乳腺癌患者的肿瘤组织及其临床资料。通过免疫组化法检测肿瘤组织中p95HER2蛋白的表达水平,比较p95HER2阳性组及阴性组的临床病理及检验资料的组间差异。通过p95HER2慢病毒质粒表达载体转染BT474细胞,构建RT-qPCR和Western blot方法验证的p95HER2蛋白稳定表达的BT474-p95细胞株。以BT474-p95细胞株为模型,经过CCK8和克隆形成试验验证考察p95HER2蛋白表达对细胞曲妥珠单抗的敏感性及其对细胞增殖的影响。结果47例HER2阳性乳腺癌患者肿瘤组织中,p95HER2的阳性率为44.7%(21/47)。p95HER2蛋白的表达情况与患者肿瘤的Ki67指标相关,差异有统计学意义(P<0.05);而p95HER2表达情况与患者的不同年龄、肿瘤大小、是否伴腋窝淋巴结转移、病理分级、雌激素受体状态比较,差异无统计学意义(P>0.05)。在血液学分析中,p95HER2蛋白的表达与患者的血清ALP、TG相关,且差异有统计学意义(P<0.05)。经过p95HER2慢病毒质粒表达载体转染的BT474细胞,可以稳定表达p95HER2蛋白,BT474-p95细胞在3.5μg/μL曲妥珠单抗药物的作用下,较BT474-NC细胞在增殖上差异有统计学意义,克隆形成也显示出相同的趋势(P<0.01)。结论在HER2阳性乳腺癌且p95HER2蛋白表达的患者肿瘤组织中,Ki67增殖指数上升,肿瘤细胞增殖能力提高,提示p95HER2的表达提高了肿瘤细胞对曲妥珠单抗治疗的内在抗性。同时,p95HER2基因不因其表达在细胞外蛋白的截短而丢失HER2蛋白促肿瘤细胞增殖的生物学功能。Objective By detecting and analyzing the expression of p95HER2 in tumor issues of HER2-positive breast cancer patients,to explore the relationship between p95HER2 and related clinical and pathological features,build p95HER2-overexpressed cells to verify its biological function in promoting cell proliferation,and provide an experimental basis for clinical HER2-positive breast therapy.Methods The tumor tissues and clinical data of 47 patients with HER2-positive breast cancer admitted to the general surgery department of Jiangsu Provincial People's Hospital from December 2020 to December 2021 were collected.The expression level of p95HER2 protein in tumor tissues was detected by immunohistochemistry and the differences between p95HER2-positive and p95HER2-negative groups were compared.BT474 cells were transfected with the p95HER2 lentiviral plasmid vector and BT474-p95 cell lines with stable expression of p95HER2 protein were constructed,which were verified by RT-qPCR and Western blot.Using the obtained BT474-p95 cell line as a model,the sensitivity of p95HER2 expression to trastuzumab and its effect on cell proliferation was investigated by CCK8 and clonal formation tests.Results In the 47 HER2-positive breast cancer patients,the positive rate of p95HER2 was 44.7%(21/47).The expression of the p95HER2 protein was correlated with the Ki67 index of the patient's tumor,and the difference was statistically significant(P<0.05).However,there was no statistically significant difference in the expression of p95HER2 compared to the patient's age,tumor size,presence of axillary lymph node metastasis,pathological grade,and estrogen receptor status(P>0.05).In hematological analysis,the expression of p95HER2 protein was correlated with patients'serum ALP and TG,and the difference was statistically significant(P<0.05).BT474 cells were transfected with the p95HER2 lentivirus plasmid vectors and BT474-p95 cells were obtained which can stably express the p95HER2 protein.When treated with 3.5μg/μL trastuzumab,cell proliferation of

关 键 词：HER2 p95HER2 HER2阳性乳腺癌 曲妥珠单抗 细胞增殖 

分 类 号：R737.9[医药卫生—肿瘤]