Balb/c裸鼠LNCaP细胞前列腺癌原位移植模型的建立及超声造影在肿瘤血管生成中的作用  

Establishment of LNCaP cell orthotopic transplantation model of prostate cancer in Balb/c nude mice and the role of contrast-enhanced ultrasound in tumor angiogenesis

作　　者：王雨豪 叶成坤 刘卫勇 Wang Yuhao;Ye Chengkun;Liu Weiyong(Wannan Medical College,Wuhu 241002,Anhui,China;Department of Neurosurgery,The First Affiliated Hospital of USTC,Hefei 230001,Anhui,China;Department of Ultrasound,The First Affiliated Hospital of USTC,Hefei 230001,Anhui,China)

机构地区：[1]皖南医学院,安徽芜湖241002 [2]中国科学技术大学附属第一医院神经外科,安徽合肥230001 [3]中国科学技术大学附属第一医院超声科,安徽合肥230001

出　　处：《右江民族医学院学报》2022年第5期644-649,680,共7页Journal of Youjiang Medical University for Nationalities

摘　　要：目的构建Balb/c裸鼠LNCaP细胞前列腺癌原位移植模型并应用超声造影(CEUS)评价肿瘤血管生成的效果。方法将LNCaP细胞接种到Balb/c裸鼠前列腺背叶中,构建Balb/c裸鼠LNCaP细胞前列腺癌原位移植模型;通过超声成像监测肿瘤的产生及体积变化情况,应用CEUS检查肿瘤内血流灌注情况并勾画感兴趣区域(ROI)生成时间强度曲线(TIC),通过仪器自动生成并记录造影剂进入肿瘤的到达时间(Atm)、到达峰值时间(TtoPk)、造影曲线下面积(AuC)等超声造影参数;通过病理学检查分析雄激素受体(AR)的表达,免疫组织化学染色检测肿瘤组织中血小板内皮细胞粘附分子(CD31)和血管内皮生长因子A(VEGF-A)的表达,利用Image J软件对免疫组织化学结果进行分析并计算CD31与VEGF-A抗体染色的平均光密度值(AOD);最后应用Prism 9.0统计软件对CEUS参数Atm、TtoPk、AuC与AOD进行Pearson相关性分析。结果肿瘤构建第50天,原位肿瘤成瘤率为71.43%(15/21),原位肿瘤平均体积为(288.81±76.49)mm 3;通过超声造影曲线分析显示Atm为(1.59±0.65)s,TtoPk为(11.59±3.68)s,AuC为(1933.91±454.38)dB;解剖发现肿瘤呈椭圆形,肿瘤组织的免疫组织化学染色结果显示:肿瘤细胞中AR染色显示强阳性,AR定位于细胞核内;免疫组织化学染色发现CD31阳性、VEGF-A阳性,Image J软件分析显示CD31的AOD=0.39±0.06、VEGF-A的AOD=0.45±0.05,Prism统计分析显示VEGF-A与Atm、Ttopk、AuC之间相关性为r=-0.934(P<0.001)、r=-0.878(P<0.001)、r=0.868(P<0.001),CD31与Atm、Ttopk、AuC之间相关性为r=-0.678(P<0.001)、r=-0.360(P=0.018)、r=0.310(P=0.031)。结论本研究通过细胞悬液注射法成功建立了LNCaP细胞原位移植模型,CEUS可以有效地评价原位前列腺肿瘤中血管的生成情况。Objective To establish a LNCaP cell orthotopic transplantation model of prostate cancer in Balb/c nude mice,and to evaluate the tumor angiogenesis by contrast-enhanced ultrasound(CEUS).Methods LNCaP cells were inoculated into the dorsal lobe of the prostate of Balb/c nude mice to construct a LNCaP cell orthotopic transplantation model of prostate cancer.Ultrasound imaging was used to monitor the generation and volume changes of tumors.CEUS was used to examine the blood perfusion in tumors and delineate the region of interest(ROI)to generate time intensity curve(TIC).CEUS parameters such as arrival time(Atm),time to peak(TtoPk)and area under the contrast-enhanced curve(AuC)were automatically generated and recorded by the instrument.The expression of androgen receptor(AR)was analyzed by pathological examination,and the expression of platelet endothelial cell adhesion molecule(CD31)and vascular endothelial growth factor A(VEGF-A)in tumor tissues was detected by immunohistochemistry.Image J software was used to analyze the results of immunohistochemistry and calculate the average optical density value(AOD)of CD31 and VEGF-A antibody staining.Finally,Pearson correlation analysis was performed on the CEUS parameters Atm,TtoPk,AuC and AOD with statistical software Prism 9.0.Results On the 50th day of tumor construction,the orthotopic tumor formation rate was 71.43%(15/21)and the average orthotopic tumor volume was(288.81±76.49)mm^(3).CEUS curve analysis showed that Atm was(1.59±0.65)s,TtoPk was(11.59±3.68)s,AuC was(1933.91±454.38)dB.The anatomy revealed that the tumor was oval in shape.Immunohistochemistry of the tumor tissue showed that AR staining was strongly positive in the tumor cells,and AR was localized in the nucleus.Immunohistochemistry staining showed that CD31 and VEGF-A were positive,and image J software analysis showed that the AOD of CD31 was 0.39±0.06,the AOD of VEGF-A was 0.45±0.05.Prism statistical analysis showed that the correlations between VEGF-A and Atm,Ttopk,AuC were r=-0.934(P<0.001),r=-0.8

关 键 词：前列腺肿瘤 原位移植模型 超声造影 血管生成 

分 类 号：R737.25[医药卫生—肿瘤]