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EP4 upregulation of Ras signaling and feedback regulation of Ras in human colon tissues and cancer cells

机译:人结肠组织和癌细胞中Ras信号的EP 4 上调和Ras的反馈调节

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Previous studies indicate that COX-2 and prostaglandin E2 (PGE2) receptor subtypes are involved in intestinal carcinogenesis and activation of downstream pathways. In this report, we try to understand the association of PGE2 receptor and K-ras cellular mechanism during the development of colorectal cancer. We collected 21 colorectal cancer patients and compared the protein expression of tumor tissues and normal mucosa tissues by using immunoblot. Furthermore, we transferred empty vector and pcDNA-K-ras into Ras-HT29 colon cancer cells. Result showed that phosphorylation of Akt and EP1/EP4 were over-expressed in the colorectal tumor tissue. K-ras induces HT29 cells proliferation through the expressions of COX-2, EP1/EP4, pAkt, GSK3β and increases Tcf transcriptional factor activation. Additionally, Ras protein was suppressed when treated with EP4 inhibitor in Ras-HT29 cell. In cell cycle assay, K-ras mutation causing cell cycle S phase was prolonged with an increase in the G2/M phase ratio. In conclusion, we suggested that Ras overexpression leads to cell proliferation through activating Ras/PI3K/GSK3β/EP4 PGE2 receptor signals and caused a feedback regulation of Ras by EP4 in colorectal tumor progression.
机译:先前的研究表明,COX-2和前列腺素E 2 (PGE 2 )受体亚型与肠癌的发生和下游通路的激活有关。在这篇报告中,我们试图了解大肠癌发展过程中PGE 2 受体与K-ras细胞机制的关系。我们收集了21名结直肠癌患者,并通过免疫印迹比较了肿瘤组织和正常黏膜组织的蛋白表达。此外,我们将空载体和pcDNA-K-ras转移到Ras-HT29结肠癌细胞中。结果表明,Akt和EP 1 / EP 4 的磷酸化在结直肠肿瘤组织中过度表达。 K-ras通过COX-2,EP1 / EP4,pAkt,GSK3β的表达诱导HT29细胞增殖,并增加Tcf转录因子的激活。另外,用EP 4 抑制剂处理Ras-HT29细胞后,Ras蛋白被抑制。在细胞周期测定中,引起K-ras突变的细胞周期S期随着G2 / M期比率的增加而延长。总之,我们认为Ras的过表达通过激活Ras / PI3K /GSK3β/ EP 4 PGE 2 受体信号导致细胞增殖,并引起EP4对Ras的反馈调节。大肠肿瘤进展。

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