Wang Na, Xu Xinzhi, Zhong Yizhe, Wan Yujie, Hong Ruixia, Wang Qizhi, Tang Jia, Gong Jiaqi, Zhou Hang, Li Fang
School of Medicine, Chongqing University Cancer Hospital, Chongqing University, Chongqing, China.
Department of Ultrasound, Chongqing University Cancer Hospital, Chongqing, China.
BMC Cancer. 2025 Apr 24;25(1):769. doi: 10.1186/s12885-025-14143-7.
Prostate cancer is one of the most prevalent malignancies in men. Once prostate cancer advances to castration-resistant prostate cancer (CRPC), the 5-year survival rate can decrease to as low as 14 months. However, the current primary diagnostic method, PSA testing, is associated with a lengthy detection cycle, limited accuracy, and delays in identifying disease progression. Consequently, there is an urgent need to develop an imaging technique that enables early and accurate diagnosis of CRPC.
First, immunofluorescence was used to verify that the expression of NRP2 on endothelial cells of neovasculature increased with the progression of prostate cancer. Next, NRP2-modified microbubbles (MBs) were prepared, and their specific targeting ability to endothelial cells was validated through parallel plate flow experiments. Subsequently, co-culture systems of prostate cancer cells and endothelial cells were established. Based on this, the proangiogenic effect of prostate cancer was systematically explored, and the differential expression of NRP2 was analyzed. A combination of immunofluorescence localization, flow cytometry, western blotting, and angiogenesis assays was used. Finally, in a subcutaneous tumor-bearing mouse model, ultrasound molecular imaging (USMI) was implemented, and the ultrasound contrast intensity of attached MBs was monitored and quantitatively analyzed.
This study confirmed the clear colocalization of NRP2 with CD31 in prostate cancer tissues. Secondly, MBs exhibited specific binding ability under dynamic conditions to microvascular endothelial cells (HMEC-1). Subsequently, with the progression of CRPC, the expression of NRP2 on HMEC-1 cells gradually increased, accompanied by a significant enhancement in their angiogenic capacity. Lastly, compared with control mice, the USMI signals in tumor-bearing mice from the hormone-sensitive prostate cancer (HSPC), non metastatic, castration-resistant prostate cancer (nmCRPC), and metastatic, castration-resistant prostate cancer (mCRPC) groups were significantly increased. This finding provides a potential new pathway for clinical diagnosis of the development of CRPC.
Regarding the progression of prostate cancer, the expression of NRP2 on neovascular endothelial cells gradually increases, potentially serving as a molecular target for early diagnosis of CRPC. The attached MBs intensity has significant differences in prostate cancer models at different stages. These findings suggest that ultrasound contrast imaging based on MBs could be a novel strategy for the early diagnosis of CRPC.
前列腺癌是男性中最常见的恶性肿瘤之一。一旦前列腺癌发展为去势抵抗性前列腺癌（CRPC），5年生存率可能降至低至14个月。然而，当前的主要诊断方法，即前列腺特异性抗原（PSA）检测，存在检测周期长、准确性有限以及识别疾病进展延迟等问题。因此，迫切需要开发一种能够早期、准确诊断CRPC的成像技术。
首先，采用免疫荧光法验证新生血管内皮细胞上神经纤毛蛋白2（NRP2）的表达随前列腺癌进展而增加。接下来，制备了NRP2修饰的微泡（MBs），并通过平行板流动实验验证了其对内皮细胞的特异性靶向能力。随后，建立了前列腺癌细胞与内皮细胞的共培养体系。在此基础上，系统地探究了前列腺癌的促血管生成作用，并分析了NRP2的差异表达。采用免疫荧光定位、流式细胞术、蛋白质免疫印迹和血管生成分析等方法相结合。最后，在皮下荷瘤小鼠模型中实施超声分子成像（USMI），监测并定量分析附着的MBs的超声造影强度。
本研究证实了NRP2与CD31在前列腺癌组织中明显共定位。其次，MBs在动态条件下对微血管内皮细胞（HMEC-1）表现出特异性结合能力。随后，随着CRPC的进展，HMEC-1细胞上NRP2的表达逐渐增加，同时其血管生成能力显著增强。最后，与对照小鼠相比，激素敏感性前列腺癌（HSPC）、非转移性去势抵抗性前列腺癌（nmCRPC）和转移性去势抵抗性前列腺癌（mCRPC）组荷瘤小鼠的USMI信号显著增加。这一发现为CRPC发展的临床诊断提供了一条潜在的新途径。
关于前列腺癌的进展，新生血管内皮细胞上NRP2的表达逐渐增加，有可能作为CRPC早期诊断的分子靶点。附着的MBs强度在不同阶段的前列腺癌模型中存在显著差异。这些发现表明基于MBs的超声造影成像可能是CRPC早期诊断的一种新策略。
