摘要
目的探讨超声造影(CEUS)对裸鼠食管癌移植瘤血管生成情况的评估价值。方法选取18只裸鼠,于其背部皮下注射人食管癌细胞株建立食管癌移植瘤模型,分别于移植后4、6、8周均随机选取6只裸鼠,先应用二维超声观察移植瘤回声、形态和边界,再行CEUS检查并获取时间-强度曲线(TIC)。然后处死所有裸鼠,使用HE染色观察移植瘤细胞结构和组织形态,免疫组化观察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),比较移植后不同时间移植瘤CEUS表现、TIC定量参数及病理学检查结果的差异。Pearson相关分析法分析TIC定量参数与VEGF蛋白表达、MVD的相关性。结果所有裸鼠食管癌移植瘤模型均成功建立,未出现死亡。移植后4周,移植瘤二维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,CEUS表现为移植瘤内部呈均匀增强;移植后6周,移植瘤二维超声表现为肿块边界欠清晰,内部回声不均匀,CEUS表现为移植瘤内部呈不均匀高增强;移植后8周,移植瘤二维超声表现为肿块边界不清晰,内部回声不均匀,CEUS表现为移植瘤内部出现灌注缺损。移植后4、6、8周移植瘤峰值强度(PI)、曲线下流入面积(AWI)及曲线下流出面积(AWO)比较,差异均有统计学意义(均P<0.05);与移植后4周比较,移植后6、8周移植瘤PI、AWI和AWO均升高(均P<0.05);与移植后6周比较,移植后8周移植瘤PI和AWO均升高(均P<0.05)。HE染色显示,移植后4周移植瘤见角化珠且细胞间见细胞间桥;移植后6周移植瘤角化珠和细胞间桥均稍减少,毛细血管增多;移植后8周,移植瘤角化珠和细胞间桥均明显减少,毛细血管增多,可见病理性核裂变。免疫组化显示,移植后4、6、8周MVD和VEGF蛋白表达比较,差异均有统计学意义(均P<0.05);与移植后4周比较,移植后6、8周移植瘤MVD均升高,移植后8周移植瘤VEGF蛋白表达升高,差异均有统计学意义(均P<0.05)。相关性分析显示,移植瘤PI、AWI、AWO与MVD、VEGF蛋白表达均呈正相关(均P<0.001)。结论CEUS能够有效观察裸鼠食管癌移植瘤内部血流灌注情况,TIC定量参数PI、AWI、AWO随着移植瘤的生长而逐渐升高,其与移植瘤MVD、VEGF蛋白表达均呈正相关,可以较好地评估移植瘤血管生成情况。
Objective To explore the evaluation value of contrast-enhanced ultrasound(CEUS)for angiogenesis of transplanted tumor in nude mice with esophageal cancer.Methods A total of 18 nude mice were selected and given subcutaneous injection of human esophageal cancer cell lines to construct models of transplanted tumor.At 4,6 and 8 weeks after transplantation,6 nude mice were randomly selected.The echo,morphology and boundary of transplanted tumor were observed by two-dimensional ultrasound,then CEUS was performed to obtain the time-intensity curve(TIC).All mice were killed at 4,6 and 8 weeks after transplantation,The cell structure and tissue morphology of transplanted tumors were observed by HE staining,the vascular endothelial growth factor(VEGF)protein expression and microvascular density(MVD)were observed by immunohistochemistry.The differences in CEUS manifestations,TIC quantitative parameters expression and pathological results of transplanted tumors were compared at different time points after transplantation.The correlation between TIC quantitative parameters and VEGF protein expression,MVD were analyzed by Pearson correlation analysis.Results All models of nude mice with transplanted tumors were successfully prepared.At 4 weeks after transplantation,two-dimensional ultrasound showed that there were ellipsoid-like hypoechoic lumps with clear boundary and uneven internal echo,while CEUS showed that there was homogeneous enhancement in transplanted tumors.At 6 weeks after transplantation,two-dimensional ultrasound showed that there was unclear boundary of lumps and uneven internal echo,while CEUS showed that there was uneven and high enhancement of blood flow signals in transplanted tumors.At 8 weeks after transplantation,two-dimensional ultrasound showed that there was unclear boundary of lumps and uneven internal echo,while CEUS showed that there were defects of blood flow signal perfusion in transplanted tumors.There were significant difference in peak intensity(PI),area wash in(AWI)and area wash out(AWO)of transplanted tumors at 4,6 and 8 weeks after transplantation(all P<0.05).Compared with those at 4 weeks after transplantation,PI,AWI and AWO were increased at 6 and 8 weeks after transplantation(all P<0.05).Compared with those at 6 weeks after transplantation,PI and AWO were increased at 8 weeks after transplantation(both P<0.05).The results of HE staining showed that at 4 weeks after transplantation,there were keratin pearls in transplanted tumors and intercellular bridges in cells.At 6 weeks after transplantation,keratin pearls and intercellular bridges were slightly decreased,and capillaries were increased.At 8 weeks after transplantation,keratin pearls and intercellular bridges were significantly decreased,capillaries were increased and there was pathological nuclear fission.There were significant differences in MVD and VEGF protein expression at 4,6 and 8 weeks after transplantation(all P<0.05).Compared with those at 4 weeks after transplantation,MVD of transplanted tumors was increased at 6 and 8 weeks after transplantation,and VEGF protein expression was increased at 8 weeks after transplantation,the differences were statistically significant(all P<0.05).Correlation analysis showed that PI,AWI and AWO were positively correlated with VEGF protein expression and MVD(all P<0.001).Conclusion CEUS can accurately observe the blood perfusion in transplanted tumors in nude mice.The TIC quantitative parameters PI,AWI and AWO gradually increase with the growth of the transplanted tumors,which are positively correlated with MVD and VEGF protein expression of transplanted tumors.It can effectively evaluate the angiogenesis of transplanted tumors in nude mice with esophageal cancer.
作者
黄浩
谢斌
张玉敏
赵现伟
陈杰能
HUANG Hao;XIE Bin;ZHANG Yumin;ZHAO Xianwei;CHEN Jieneng(Department of Ultrasound Medicine,Huanggang Central Hospital,Hubei 438000,China)
出处
《临床超声医学杂志》
CSCD
2024年第3期184-189,共6页
Journal of Clinical Ultrasound in Medicine
关键词
超声检查
造影剂
时间-强度曲线
食管癌
移植瘤
血管生成
裸鼠
Ultrasonography
Contrast agent
Time-intensity curve
Esophageal cancer,transplanted tumor
Angiogenesis
Nude mice
