超声造影时间-强度曲线评价裸鼠食管癌移植瘤血管生成情况的研究

目的 探讨超声造影(CEUS)时间-强度曲线(TIC)对裸鼠食管癌移植瘤血管生成情况的评价效果。方法 选取18只裸鼠并于背部皮下注射人食管癌细胞株建立食管癌移植瘤模型,分别于移植瘤移植后4周、6周、8周时,选取6只裸鼠,使用CEUS检查选择感兴趣区行TIC分析,比较移植后不同时间点食管癌移植瘤CEUS TIC定量参数和影像学特点,苏木素－伊红(HE)染色观察移植瘤病理变化,免疫组化分析移植瘤组织血管内皮生长因子(VEGF)蛋白表达和微血管密度(MVD),Pearson分析CEUS TIC定量参数与移植瘤MVD、VEGF相关性。结果 与移植后4周时比较,移植后6周、8周的峰值强度(PI)、曲线下流入面积(AWI)和曲线下流出面积(AWO)均升高(P<0.05);与移植后6周时比较,移植后8周的PI和AWO均升高(P<0.05)。移植后4周时食管癌移植瘤的常规二维灰阶超声表现为类椭圆形低回声肿块、边界清晰、内部回声不均匀,且多普勒血流显像显示裸鼠移植瘤体内和周边见少许血流信号;移植后6周时食管癌移植瘤内部显示灌注不均匀增强,移植后8周时食管癌移植瘤内部出现灌注缺损。HE染色结果显示,在移植4周时移植瘤有角化珠且细胞间有细胞间桥,随着移植后时间延长角化珠和细胞间桥逐渐减少。与移植后4周时比较,移植后6周、8周的MVD均升高(P<0.05),移植后8周的VEGF蛋白表达升高(P<0.05)。Pearson分析显示裸鼠人食管癌移植瘤VEGF蛋白表达、MVD均与PI、AWI、AWO呈正相关(P<0.001)。结论 CEUS技术TIC分析可以有效评价裸鼠人食管癌移植瘤血管生成情况。

Evaluation on angiogenesis of esophageal transplanted tumor in nude mice by contrast-enhanced ultrasound time-intensity curves

Objective To explore the evaluation effect of contrast-enhanced ultrasound (CEUS) time-intensity curves (TIC) on angiogenesis of esophageal transplanted tumor in nude mice. Methods A total of 18 nude mice were selected and given subcutaneous injection of human esophageal cancer cell lines to construct models of esophageal transplanted tumor. At 4, 6 and 8 weeks after transplantation, 6 nude mice were selected for TIC analysis in region of interest (ROI) by CEUS. Quantitative parameters of CEUS TIC and imaging characteristics in esophageal transplanted tumor were compared after transplantation. The pathological changes of transplanted tumors were observed by hematoxylin-eosin (HE) staining, the expression of vascular endothelial growth factor (VEGF) protein and microvascular density (MVD) were analyzed by immunohistochemistry. The correlation between quantitative parameters of CEUS TIC and MVD, VEGF was analyzed by Pearson correlation analysis. Results Compared with those at 4 weeks after transplantation, peak intensity (PI), area wash in (AWI) and area wash out (AWO) were increased at 6 and 8 weeks after transplantation (P<0.05). Compared with those at 6 weeks after transplantation, PI and AWO were increased at 8 weeks after transplantation (P<0.05). At 4 weeks after transplantation, routine two-dimensional gray scale ultrasound showed that there were ellipsoid-like hypoechoic lumps with clear boundary and uneven internal echo. Color Doppler flow imaging showed that there were a few of blood flow signals in and around transplanted tumors of nude mice. At 6 weeks after transplantation, there was uneven and enhanced perfusion in transplanted tumors, and there were perfusion defects at 8 weeks after transplantation. The results of HE staining showed that at 4 weeks of transplantation, there was keratin pearl in transplanted tumors and intercellular bridge in cells. With time, keratin pearl and intercellular bridge were gradually decreased. Compared with that at 4 weeks after transplantation, MVD was increased at 6 and 8 weeks after transplantation (P<0.05), and expression of VEGF protein was increased at 8 weeks after transplantation (P<0.05). Pearson analysis showed that VEGF protein and MVD were positively correlated with PI, AWI and AWO (P<0.001). Conclusion CEUS TIC analysis can effectively evaluate the angiogenesis of human esophageal transplanted tumors in nude mice.