Substantial changes in gene expression of Wnt, MAPK and TNFα pathways induced by TGF-β1 in cervical cancer cell lines
Substantial changes in gene expression of Wnt, MAPK and TNFα pathways induced by TGF-β1 in cervical cancer cell lines
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DOI：
10.1093/carcin/bgi110
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发表时间：
2005-09-01
期刊：
影响因子：
4.7
通讯作者：
Gorter, A
中科院分区：
文献类型：
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作者：
Kloth, JN;Fleuren, GJ;Gorter, A
转化生长因子-β1(TGF-β1)是一种强有力的上皮细胞增殖抑制因子。然而，在宫颈癌的发展过程中，伴随着转化生长因子-β1产生的增加伴随着对转化生长因子-β1生长限制作用的敏感性降低。转化生长因子-β1对免疫系统的细胞具有抗增殖作用，因此有利于肿瘤的进展。本研究的目的是在对转化生长因子-β1具有不同敏感性的宫颈癌细胞系中，研究转化生长因子-β1对细胞生长和死亡途径中基因表达谱的影响。为此，我们研究了转化生长因子-β1刺激的高(CC10B)、中(SIHA)和低(HeLa)对转化生长因子-β1抗增殖作用敏感的宫颈癌细胞株在0、6、12和24小时的基因表达变化。在我们的研究中，我们发现了与肿瘤坏死因子α(TNFα)、丝裂原活化蛋白激酶(MAPK)和无翼型(WNT)通路有关的新的靶基因，以响应转化生长因子-β1。在对转化生长因子-β1敏感和不敏感的细胞系中，观察到了涉及肿瘤坏死因子α、MAPK、Wnt和Smad通路的基因表达的显著差异。由于这些途径与细胞增殖和细胞死亡有关，因此这些途径可能在决定细胞对转化生长因子-β1诱导的细胞生长抑制的整体敏感性方面发挥作用。结果随后通过实时定量聚合酶链式反应进行了验证。通过酶联免疫吸附试验检测，细胞对转化生长因子-β1诱导的生长抑制的抵抗力增加与细胞系产生转化生长因子-β1的增加相关。转化生长因子-β1的产生不会抑制细胞的生长，因为用抗转化生长因子-β1抗体阻断转化生长因子-β1蛋白对细胞的增殖没有影响。肿瘤细胞分泌的转化生长因子-β1更可能有助于旁分泌刺激肿瘤的发展。
Transforming growth factor-beta 1 (TGF-beta 1) is a potent inhibitor of epithelial cell proliferation. During the development of cervical carcinoma however, an increase in production of TGF-beta 1 is accompanied by decreased sensitivity for the growth-limiting effect of TGF-beta 1. TGF-beta 1 has an anti-proliferative effect on cells of the immune system and thus can be advantageous for tumor progression. The aim of the present study was to determine the effect of TGF-beta 1 on mRNA expression profile of genes in pathways involved in cell growth and cell death, in cervical carcinoma cell lines with different sensitivity to TGF-beta 1. For this purpose, we have investigated changes in gene expression in TGF-beta 1 stimulated cervical cancer cell lines with high (CC10B), intermediate (SiHa) and low (HeLa) sensitivity to the anti-proliferative effect of TGF-beta 1, at timepoints 0, 6, 12 and 24 h. Microarray analysis, using Affymetrics focus arrays, representing 8973 genes, was used to measure gene expression. In our study novel target genes involved in tumor necrosis factor alpha (TNF alpha), mitogen-activated protein kinase (MAPK) and wingless type (Wnt) pathways in response to TGF-beta 1 were found. Substantial differences in gene expression between TGF-beta 1 sensitive and insensitive cell lines were observed involving genes in TNF alpha, MAPK, Wnt and Smad pathways. Since these pathways are implicated in cell proliferation and cell death, these pathways may play a role in determining the overall sensitivity of a cell to TGF-beta 1 induced cell growth inhibition. The results were subsequently validated by quantitative real-time PCR. Increased resistance to TGF-beta 1 induced cell growth inhibition was correlated with an elevated production of TGF-beta 1 by the cell lines, as measured by enzyme linked immunosorbent assay. TGF-beta 1 production did not inhibit cell growth, since blocking TGF-beta 1 protein by anti-TGF-beta had no effect on cell proliferation. TGF-beta 1 excretion by tumor cells more likely contributes to paracrine stimulation of tumor development.