Angiogenesis Research in Mouse Mammary Cancer Based on Contrast-enhanced Ultrasonography: Exploratory Study

Abstract

The objective of this study was to investigate the contrast-enhanced ultrasound (CEUS) characteristics of tumor angiogenesis in mouse mammary cancer. Twenty-four mice were examined with ultrasound and CEUS at 2-12 days after implantation. Four to five mice were assessed daily, and one to three mice were then sacrificed for histology. All of the histologic slides were reviewed and correlated with CEUS findings. A total of 46 cases of ultrasound examination had been performed in 24 mice. The mice were classified into three groups according to the tumor growth: group 1 (2~6 days after implantation, n = 20 cases), group 2 (7~9 days after implantation, n = 15 cases), and group 3 (10~12 days after implantation, n = 11 cases). In group 1, all tumors presented as a homogeneous hypoechoic mass with no color Doppler signals. However, three CEUS patterns were observed: 14 tumors presented as type I (peripheral ring enhancement with no enhancement within the tumor), 4 tumors presented as type II (peripheral ring enhancement with deep penetration), and 2 tumors presented as type III (homogeneous or heterogeneous enhancement in the entire tumor). In group 2, there was only difference in the echo (heterogeneous or not) and color Doppler signals (with or without) among the tumors in conventional ultrasound, but four CEUS patterns were observed and most presented as type III (53.3%, 8/15). In group 3, most tumors presented as a heterogeneous solid mass (81.8%, 9/11) with color signals (100%, 11/11), and almost all tumors presented as enhancement of type IV (peripheral ring enhancement with focal nodular enhancement) (90.9%, 10/11).The histologic results showed that the enhanced areas mainly corresponded to tumor cells, large tortuous vessels, and an inflammatory cell infiltrate. Nonenhanced areas corresponded to large areas of necrotic tissue or tumor cells, which arranged loosely with the small zone of necrosis. CEUS could image the progression of vessel formation. Moreover, most importantly, CEUS is able to identify angiogenesis before the change of tumor color Doppler, and presents different enhanced patterns at different tumor growth times, which corresponded to tumor histologic features.