摘要
目的:探讨细胞活素肿瘤坏死因子-α(TNF-α)、SNAP(S-Nitroso-Nacetyl-penicillamine)对胰腺癌细胞产生血管内皮生长因子A、C(VEGF-A、C)的调节。方法:用Northern杂交和Western杂交法分析6种人胰腺癌细胞株中VEGF-A、C基因和蛋白的表达;以TNF-α或SNAP刺激其中两个细胞株后用逆转录-聚合酶链式反应技术(RT-PCR)分析其VEGF-A、C基因的表达。结果:Northern杂交法显示这6种胰腺癌细胞株均有4.1kbVEGF-A基因和2.4kbVEGF-C基因的表达;Western杂交法显示它们均有分子量为43kD的VEGF-A蛋白质和分子量为55kD的VEGF-C蛋白质的表达。RT-PCR分析法显示:TNF-α使细胞株COLO-357产生VEGF-A、VEGF-CmRNA分别减少约1~2.5倍、1~2倍,使细胞株CAPAN-1产生VEGF-A、VEGF-CmRNA分别减少约1倍、1.6~2.5倍;而SNAP刺激细胞株COLO-357产生VEGF-AmRNA增加约5倍,刺激细胞株CAPAN-1产生VEGF-AmRNA增加约4倍,但对这两种细胞株产生VEGF-CmRNA均无明显刺激作用。结论:细胞活素TNF-α和低氧通过调节血管内皮生长因子A、C的表达而影响胰腺癌细胞的生物学特性,抑制癌细胞的增殖,促进其凋亡、死亡或进展、恶化。
Objective: To explore cytokines tumor necrosis factor-alpha (TNF-α) and S-Nitroso-Nacetyl-penicillamine (SNAP, nitrix oxide donor, leads to hypoxia state in the culture medium.) regulate to expression of vascular endothelial growth factor-A, C (VEGF-A, C) in pancreatic cancer cells. Methods: The method of Northern blot and Western blot was respectively used to analyze the expression of gene and protein of VEGF-A, C in six human pancreatic cancer cells lines and the reversetranscription polymerase chain reaction (RT-PCR) method was used to analyze VEGF-A, C gene expression in two tested cell lines under the stimulation with TNF-α or SNAP. Results: Northern blot analysis revealed the presence of the 4.1-kb VEGF-A mRNA transcript and 2.4-kb VEGF-C mRNA transcript in all six tested cell lines; Western blot analysis revealed the presence of Mr 43-kDa VEGFA protein and Mr 55-kDa VEGF-C protein in all the cell lines. RT-PCR analysis showed that by stimulation with TNF-α, there were a 1- to 2.5-fold and 1- to 2-fold decrease in COLO-357 cell line, and a 1-fold and 1.6- to 2.5- fold decrease in CAPAN-1 cell line in the level of VEGF-A and of VEGF-C mRNA, respectively; The level of VEGF-A mRNA increased five times in COLO-357 cell line and four times in CAPAN-1 cell line by stimulation with SNAP, however, no effect was found in the two cell lines by stimulation with SNAP. Conclusions: Cytokines TNF-α and SNAP affect the biological character of pancreatic cancer cells by regulating the expression of VEGF-A, VEGF-C, and promote the apotosis and death or the growth and distant metastasis of pancreatic cancer cells.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2005年第15期852-855,共4页
Chinese Journal of Clinical Oncology
基金
教育部留学回国人员科研启动基金项目资助(编号:教外司留[2002]247号)
关键词
胰腺肿瘤
血管内皮生长因子A、C
肿瘤坏死因子
低氧
Pancreatic neoplasms Vascular endothelial growth factor-A, -C Tumor necrosis factor Hypoxia