Vascular endothelial growth factor in salivary pleomorphic adenomas: one of the reasons for their poorly vascularized stroma
Vascular endothelial growth factor in salivary pleomorphic adenomas: one of the reasons for their poorly vascularized stroma
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DOI：
10.1007/s00428-005-1219-1
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发表时间：
2005-06-01
期刊：
影响因子：
3.5
通讯作者：
Saku, T
中科院分区：
文献类型：
--
作者：
Swelam, W;Ida-Yonemochi, H;Saku, T
为了更好地了解多形性腺瘤间质血管化不良的背景，我们试图确定多形性腺瘤中血管和缺氧条件相关分子的表达。手术标本和涎腺多形性腺瘤原代培养的肿瘤细胞用于CD 31、血管内皮生长因子（VEGF）及其受体Flk-1和Flt-1的免疫组化，以及缺氧标志物，如缺氧诱导因子-1 α（HIF-1 α）和乳酸脱氢酶-1（LDH）。同时，通过逆转录聚合酶链反应（RT-PCR）和PCR产物的直接测序分析手术标本中VEGF基因的选择性剪接模式和HIF-1 α基因的表达水平。除了与CD 31+血管内皮细胞共免疫定位外，VEGF及其受体在正常导管上皮细胞和肌上皮细胞以及导管结构和粘液软骨样基质中的肿瘤细胞中也有表达。HIF-1 α和LDH的免疫定位在VEGF阳性区域得到证实。在培养的多形性腺瘤细胞中证实了VEGF和其他免疫荧光信号。RT-PCR结果显示VEGF基因至少有4种剪接方式，其中VEGF（121）剪接方式增强最明显，多形性腺瘤中HIF-1 α表达水平较高。结果表明，多形性腺瘤细胞产生VEGF在几个功能形式为自己的增殖或分化，和VEGF的表达是由缺氧的情况下，血管化不良的多形性腺瘤控制。
To better understand the poorly vascularized background of the stroma of pleomorphic adenomas, we attempted to determine the expression of molecules related to blood vessels and hypoxic conditions in pleomorphic adenoma. Surgical specimens and tumor cells in primary culture of salivary pleomorphic adenomas were used for immunohistochemistry for CD31, vascular endothelial growth factor (VEGF) and its receptors Flk-1 and Flt-1, as well as for hypoxia markers, such as hypoxia-inducible factor-1 alpha (HIF-1 alpha) and lactate dehydrogenase-1 (LDH). At the same time, alternative splicing modes of the VEGF gene and expression levels of the HIF-1 alpha gene were analyzed in surgical specimens by means of reverse-transcription polymerase chain reaction (RT-PCR) and direct sequencing of the PCR products. In addition to co-immunolocalization with CD31+ vascular endothelial cells, VEGF and its receptors were demonstrated in normal duct epithelial and myoepithelial cells as well as in tumor cells in ductal structures and in myxochondroid stromata. Immunolocalizations for HIF-1 alpha and LDH were confirmed in the VEGF-positive area. Immunofluorescence signals for VEGF and others were confirmed in pleomorphic adenoma cells in culture. RT-PCR results showed that there were at least four splicing modes of the VEGF gene, among which VEGF(121) was most enhanced, and higher HIF-1 alpha levels in pleomorphic adenomas. The results suggest that pleomorphic adenoma cells produce VEGF in several functional forms for their own proliferation or differentiation, and that the VEGF expression is controlled by hypoxic circumstances of poorly vascularized pleomorphic adenomas.