klf6基因在舌癌细胞诱导分化过程中的作用分析-role of kl f6 gene in differentiation of tongue cancer cells.docxVIP

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klf6基因在舌癌细胞诱导分化过程中的作用分析-role of kl f6 gene in differentiation of tongue cancer cells.docx

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klf6基因在舌癌细胞诱导分化过程中的作用分析-role of kl f6 gene in differentiation of tongue cancer cells

PAGE PAGE VI III III KLF6、p53、c-Jun 及 Cyclin D1 蛋白表达的变化，显示 KLF6 蛋白和 p53 在 HMBA 处理后表达显著上调（p0.05），而 c-Jun 和 Cyclin D1 则表达显著下调（p0.05）。结论：HMBA 对人舌癌细胞 TCA 8113 具有抑制增殖、促进分化和诱导凋亡的作用；探明了 HMBA 对人舌癌细胞诱导分化处理可上调 KLF6 mRNA，增加 KLF6 蛋白的表达，上调 p53，下调 c-Jun 和 Cyclin D1，其作用机理一方面可能通过 p53 上调 p21 的表达和活性，另一方面可能通过 KLF6 解除 c-Jun 对 p21 的抑制作用，下调 Cyclin D1，抑制 Cdk4/6 和 Cyclin D1 复合物活性，将人舌癌细胞阻滞于 GO/G1 期，诱导人舌癌细胞向正常细胞分化，恢复正常细胞的表型和功能，并促进细胞的凋亡。关键词 KLF6；HMBA；诱导分化；TCA 8113 RESEARCH ON THE MECHANISMS BASED ON KLF6 GENE OF INDUCING TCA 8113 CELLS TO DIFFERENCIATION ABSTRACT Objective To understand the role and mechanisms based on KLF6 gene of inducing TCA 8113 cells to differenciation at cellular and molecular level, and to discuss the possibility of KLF6 as a new specific target anti- cancerization of tongue cells. Methods The growth and cell morphology of TCA8113 cells were observed by MTT and under microscope .The expression of KLF6 mRNA were detect by RT-PCR. The expression of KLF6 protein,p53, c-Jun and CyclinD1 were detect by immunohistochemistry. Results After inducing by HMBA,the number of adherent cells decreased obviously along with the concentration of HMBA. The growth inhibition on TCA 8113 cells was in a concentration-time effect relationship by MTT after treating by HMBA. That means the effect of inhibition increased gradually along with the concentration of inducing agent increases, and the time extended. There were some reversal features of developing to normal cells morphologically after induced by HMBA, such as:cell morphology convergence, cell volume increased obviously, and spread out, the proportion of nucleus and cytoplasm decreased, smaller nucleus, the shape of nuclear in most cells became oval, the number of nucleolus reduced obviously,cytoplasm was abundant, the color of total cells uniformed by HE stain and so on. The cell cycle was analysed by flow cytometry. The rate of G1 phase cells was 52.0% before treating by HMBA. The rate of S phase cells was 34.0%, and 14.0