Zhang Jiali, Peng Bin
Key Lab for Oral Biomedical Engineering of Ministry of Education, School and Hospital of Stomatology, Wuhan University, China.
BMC Cancer. 2007 Jun 1;7:95. doi: 10.1186/1471-2407-7-95.
Adenoid cystic carcinoma is a high malignant carcinoma characterized by intensive local invasion and high incidence of distant metastasis. Although many reports have demonstrated that angiogenesis has played an important role in tumor metastasis, the relationship between metastasis characters and angiogenesis ability in high and low metastasis cell lines of Adenoid cystic carcinoma has rarely been reported. The present study aimed to compare the angiogenesis ability of ACC-M (high metastasis) and ACC-2 (low metastasis) cell lines in vitro. Furthermore, the activity of nuclear factor kappaappa B and the expression of vascular endothelial growth factor (VEGF) in ACC-2 and ACC-M were also detected.
Electrophoretic mobility shift assay was used to detect nuclear factor kappaappa B activity. Semi-quantitative RT-PCR was used to quantify the mRNA level of VEGF. Immuofluorescence double staining and semi-quantitative confocal laser scanning analysis was carried out to detect nuclear factor kappaappa B nuclear localization and staining intensity of VEGF. The angiogenesis ability of ACC-M and ACC-2 was compared by an in vitro three-dimensional angiogenic model assay. The vector transfection assay was performed to transfect the PCMV-IkappaBalphaM vector into ACCs cell lines expressing the phosphorylation defective IkappaBalphaM.
Nuclear factor kappaappa B activity and the rate of nuclear factor kappaappa B nuclear localization in ACC-M was significantly higher than that in ACC-2. Moreover, ACC-M exhibited higher mRNA and protein levels of vascular endothelial growth factor than ACC-2. VEGF mRNA expression was effectively decreased by inhibition of nuclear factor kappaappa B activity. Furthermore, ACC-M could remarkably stimulate the migration and tube formation of endothelial cells and induce The umbilical vein endothelial cells sprouting into the gel matrix.
These results implicated that ACCs cells with higher metastasis feature might present greater angiogenesis ability.
腺样囊性癌是一种恶性程度高的癌症，其特点是局部侵袭性强，远处转移发生率高。尽管许多报道表明血管生成在肿瘤转移中起重要作用，但腺样囊性癌高低转移细胞系中转移特征与血管生成能力之间的关系鲜有报道。本研究旨在比较腺样囊性癌高转移细胞系ACC-M和低转移细胞系ACC-2的体外血管生成能力。此外，还检测了ACC-2和ACC-M中核因子κB的活性及血管内皮生长因子（VEGF）的表达。
采用电泳迁移率变动分析检测核因子κB活性。采用半定量逆转录-聚合酶链反应定量VEGF的mRNA水平。进行免疫荧光双染色和半定量共聚焦激光扫描分析，以检测核因子κB的核定位及VEGF的染色强度。通过体外三维血管生成模型试验比较ACC-M和ACC-2的血管生成能力。进行载体转染试验，将PCMV-IκBαM载体转染至表达磷酸化缺陷型IκBαM的腺样囊性癌细胞系中。
ACC-M中核因子κB活性及核因子κB核定位率显著高于ACC-2。此外，ACC-M的血管内皮生长因子mRNA和蛋白水平高于ACC-2。抑制核因子κB活性可有效降低VEGF mRNA表达。此外，ACC-M可显著刺激内皮细胞的迁移和管腔形成，并诱导脐静脉内皮细胞向凝胶基质中发芽。
这些结果表明，具有较高转移特征的腺样囊性癌细胞可能具有更强的血管生成能力。
