Lan C Y, Ling B, Guo W W, Yin W, Zhong X G, Han Y M, Dong X F
Department of Ultrasound, the People's Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China.
Department of Pathology, the People's Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China.
Zhonghua Zhong Liu Za Zhi. 2018 Feb 23;40(2):105-109. doi: 10.3760/cma.j.issn.0253-3766.2018.02.005.
To detect the possible molecular mechanisms of the formation of vessels that encapsulated tumor clusters (VETC) and identify the relationship between vimentin protein expression in endothelial cells and contrast-enhanced ultrasound characters in VETC (+ ) hepatocellular carcinoma (HCC). A total of 64 paraffin embedded HCC tissue samples were collected, all of which the tumor diameters were between 2 cm and 5 cm measured by the preoperative ultrasound. Immunohistochemistry staining for CD34 was used to detect the formation of VETC and the expressions of angiopoietin-2 (Ang-2) and vimentin were also determined. Human umbilical vein endothelial cells (HUVECs) were treated with 150 ng/ml recombinant human Ang-2 protein (rhAng-2) at various times and the protein expression of vimentin was detected by western blot assay. The contrast-enhanced ultrasound characters were also analyzed in both VETC (+ ) and VETC (-) HCC. Tumor clusters encapsulated by vessels to form cobweb-like networks, which were identified as VETC phenotype, were observed in 27 HCC tissues (42.18%). In VETC (+ ) HCC tissues, Ang-2 was overexpressed in tumor cells and endothelial cells while vimentin was only upregulated in endothelial cells. With the treatment of 150 ng/ml rhAng-2 protein, the expression of vimentin in HUVECs was 0.878±0.102 and 0.918±0.092 at 12 h and 36 h, significantly upregulated when compared to the 0.322±0.061 at 6 h (<0.01). In contrast-enhanced ultrasound, a crack and tendon-like filling character was observed in VETC (+ ) HCC during the arterial-phase, while the large scale and diffuse-like filling character was observed in VETC (-) HCC. The filling time of unit diameter in VETC (+ ) HCC was (3.95±0.22)s, significantly longer than (2.28±0.27)s of VETC (-) HCC (<0.01). The overexpressions of Ang-2 and vimentin are positively correlated with the formation of VETC and considered as potential therapeutic targets of VETC (+ ) HCC. The crack and tendon-like filling characters in arterial-phase of contrast-enhanced ultrasound indicates the VETC (+ ) HCC.
为探究包裹肿瘤细胞簇的血管（VETC）形成的可能分子机制，并确定内皮细胞中波形蛋白的蛋白表达与VETC（+）肝细胞癌（HCC）中超声造影特征之间的关系。共收集64例石蜡包埋的HCC组织样本，所有样本术前超声测量肿瘤直径均在2 cm至5 cm之间。采用CD34免疫组化染色检测VETC的形成，并检测血管生成素-2（Ang-2）和波形蛋白的表达。用150 ng/ml重组人Ang-2蛋白（rhAng-2）在不同时间处理人脐静脉内皮细胞（HUVECs），通过蛋白质印迹法检测波形蛋白的蛋白表达。对VETC（+）和VETC（-）HCC的超声造影特征也进行了分析。在27例HCC组织（42.18%）中观察到血管包裹肿瘤细胞簇形成蜘蛛网样网络，被确定为VETC表型。在VETC（+）HCC组织中，Ang-2在肿瘤细胞和内皮细胞中均过度表达，而波形蛋白仅在内皮细胞中上调。用150 ng/ml rhAng-2蛋白处理后，HUVECs中波形蛋白在12 h和36 h时的表达分别为0.878±0.102和0.918±0.092，与6 h时的0.322±0.061相比显著上调（<0.01）。在超声造影中，VETC（+）HCC在动脉期观察到裂缝状和条索状填充特征，而VETC（-）HCC观察到大片状和弥漫状填充特征。VETC（+）HCC单位直径的填充时间为（3.95±0.22）s，显著长于VETC（-）HCC的（2.28±0.27）s（<0.01）。Ang-2和波形蛋白的过度表达与VETC的形成呈正相关，被认为是VETC（+）HCC的潜在治疗靶点。超声造影动脉期的裂缝状和条索状填充特征提示VETC（+）HCC。
