超声靶向微泡破坏介导的整合素亚基α3转染对乳腺癌生长和上皮间质转化的影响  

Influence of the growth and epithelial-mesenchymal transformation of breast cancer mediated by integrin subunitα3 transfection with ultrasound targeted microbubble destruction

作　　者：钱昭高 李在丽 QIAN Zhaogao;LI Zaii(Department of Ultrasound,the People’s Hospital of Pingyang,Zhejiang 325400,China)

机构地区：[1]平阳县人民医院超声科,浙江省温州市325400 [2]温州医科大学附属瑞安医院超声科

出　　处：《临床超声医学杂志》2024年第7期529-536,共8页Journal of Clinical Ultrasound in Medicine

基　　金：2022年度温州市基础性公益科研项目(2022Y0156)。

摘　　要：目的探讨超声靶向微泡破坏(UTMD)介导的整合素亚基α3(ITGA3)过表达质粒转染对乳腺癌生长和上皮间质转化(EMT)的影响。方法基于GEPIA平台分析TCGA数据库中乳腺癌样本的ITGA3表达情况,绘制生存曲线评估ITGA3表达水平对乳腺癌患者生存的影响。将乳腺癌BT-549细胞分为LIP-NC组(脂质体介导的阴性对照质粒转染细胞)、LIP-ITGA3组(脂质体介导的ITGA3过表达质粒转染细胞)、UTMD-NC组(UTMD介导的阴性对照质粒转染细胞)和UTMD-ITGA3组(UTMD介导的ITGA3过表达质粒转染细胞),UTMD-NC组和UTMD-ITGA3组细胞在转染过程中均接受超声辐照(频率1 MHz,声强0.75 W/cm2,时间45 s)。于扫描电子显微镜下观察LIP-NC组、UTMD-NC组细胞形态;分别应用MTT法、划痕实验、Transwell实验检测各组细胞活力、迁移能力、侵袭能力;应用Western blot法分析BT-549细胞中ITGA3和EMT、STAT3通路相关基因蛋白的表达情况。通过裸鼠皮下肿瘤细胞注射法建立裸鼠乳腺癌异种移植瘤模型,移植35 d后处死裸鼠剥离异种移植瘤并称重;应用免疫组化检测各组异种移植瘤组织中ITGA3、N-cadherin和PCNA的表达情况。结果生物信息学分析显示,乳腺癌样本中ITGA3蛋白相对表达量明显低于正常乳腺样本,差异有统计学意义(P<0.05);ITGA3低表达与乳腺癌患者不良预后相关(HR=0.81,P=0.00072)。体外实验结果显示,扫描电子显微镜下LIP-NC组细胞呈球形,细胞膜表面无明显的凹坑或孔隙;UTMD-NC组细胞形态未见改变,但细胞膜表面可见粗糙区域和小凹坑。UTMD-ITGA3组、LIP-ITGA3组细胞活力、迁移能力、侵袭能力均分别较各自对照组(UTMD-NC组、LIP-NC组)降低,差异均有统计学意义(均P<0.05);且UTMD-ITGA3组细胞活力、迁移能力、侵袭能力均较LIP-ITGA3组更低(均P<0.05)。LIP-ITGA3组ITGA3蛋白相对表达量较LIP-NC组增高,UTMD-ITGA3组ITGA3蛋白相对表达量较UTMD-NC组、LIP-ITGA3组均增高,差异均有统计Objective To investigate the influence on the growth and epithelial-mesenchymal transformation(EMT)of breast cancer by integrin subunitα3(ITGA3)transfection mediated with ultrasound targeted microbubble destruction(UTMD).Methods The expression of ITGA3 in breast cancer samples from the TCGA database was analyzed based on the GEPIA platform,and survival curves were drawn to evaluate the impact of ITGA3 expression on the survive of breast cancer patients.Breast cancer BT-549 cells were divided into LIP-NC group(liposome mediated negative control plasmid),LIP-ITGA3 group(liposome mediated ITGA3 overexpression plasmid transfected cells),UTMD-NC group(UTMD-mediated negative control plasmid transfected cells)and UTMD-ITGA3 group(UTMD-mediated ITGA3 overexpressed plasmid transfected cells).Cells in UTMD-NC and UTMD-ITGA3 groups were accepted ultrasonic irradiation(frequency 1 MHz,sound intensity 0.75 W/cm2,duration 45 s)during transfection.Cell morphology of UTMD-NC and UTMD-ITGA3 groups was observed by scanning electron microscopy.Cell viability was detected by MTT assay.Cell migration ability was detected by scratch assay,and cell invasion ability was detected by Transwell assay.Western blot analysis was performed to quantify the expression of ITGA3 and EMT-and STAT3 pathway-related proteins in BT-549 cells.A xenograft tumor model of breast cancer was established by subcutaneously injecting tumor cells into nude mice.After 35 d of transplantation,the nude mice were sacrificed and the xenograft tumors were harvested and weighed.Immunohistochemistry was used to detect the expression of ITGA3,N-cadherin and PCNA of xenograft tumor tissues in each group.Results Bioinformatics analysis showed that the expression of ITGA3 in breast cancer samples was significantly lower than that in normal breast samples(P<0.05).The low expression of ITGA3 was associated with the poor prognosis in breast cancer patients(HR=0.81,P=0.00072).In vitro experiment showed that cells in the LIP-NC group were spherical under scanning electron micro

关 键 词：超声靶向微泡破坏 乳腺癌 脂质体 整合素亚基α3 

分 类 号：R445.1[医药卫生—影像医学与核医学] R737.9[医药卫生—诊断学]