Department of Nutritional Toxicology, Institute of Nutrition, Friedrich-Schiller-University Jena, Jena, Germany,
Genes Nutr. 2012 Apr;7(2):235-46. doi: 10.1007/s12263-011-0254-6. Epub 2011 Oct 19.
Due to protection of oncogenic proteins from degradation and enhancement of glycolytic phosphometabolites for synthetic processes, respectively, heat shock protein 90 (HSP90) and pyruvate kinase type M2 (PKM2) are important proteins for tumor growth. The present study was undertaken to investigate the susceptibility of both proteins and their encoding genes to the chemopreventive agent butyrate in human colon cells. Matched tissue of different transformation stages derived from 20 individual colon cancer patients was used for the experiments. The results of quantitative real-time PCR revealed a moderate increase of HSP90β and PKM2 mRNA in colon tumors (P < 0.01) compared to normal tissues without relation to clinical parameters. The expression pattern could be confirmed for PKM2 protein by Western blot but not for HSP90β. During culturing with butyrate, the amount of PKM2 transcripts decreased in all three tissue types with the strongest effects observed in tumors (median fold decrease 45%, P < 0.05). The protein data have not reflected this influence supposing a more gradual degradation rate due to a longer half-life of PKM2. In contrast, the mRNA expression of HSP90β in normal tissue was found 1.38-fold increased by butyrate (P < 0.05), but not the corresponding protein level. HSP90β expression in adenomas and tumors remained generally insensitive. Only in malignant tissue, however, a significant correlation was found between the individual effects observed on gene and protein expression level. In conclusion, the present study identified PKM2 as a potential direct target of butyrate in neoplastic colon tissue, whereas HSP90β is none of it.
由于热休克蛋白 90(HSP90)和丙酮酸激酶 M2(PKM2)分别能保护致癌蛋白不被降解并增强糖酵解磷酸代谢物以供合成代谢使用,它们都是肿瘤生长的重要蛋白。本研究旨在探讨这两种蛋白及其编码基因对化学预防剂丁酸盐的敏感性,所用细胞为人结肠细胞。实验中使用了来自 20 名个体结肠癌患者的不同转化阶段的配对组织。实时定量 PCR 的结果显示,与正常组织相比,结肠肿瘤中 HSP90β和 PKM2 mRNA 有适度增加(P<0.01),但与临床参数无关。Western blot 可证实 PKM2 蛋白的表达模式,但不能证实 HSP90β 的表达模式。在丁酸盐培养过程中,所有三种组织类型的 PKM2 转录物含量均减少,肿瘤中观察到的影响最强(中位数下降 45%,P<0.05)。由于 PKM2 的半衰期较长,其蛋白数据并未反映出这种影响,假设其降解率更为渐进。相比之下,丁酸盐使正常组织中 HSP90β 的 mRNA 表达增加了 1.38 倍(P<0.05),但蛋白水平没有增加。腺瘤和肿瘤中 HSP90β 的表达通常不敏感。然而,只有在恶性组织中,基因和蛋白表达水平上观察到的个体效应之间才存在显著相关性。总之,本研究鉴定出 PKM2 是肿瘤性结肠组织中丁酸盐的潜在直接靶标,而 HSP90β 则不是。