鼻咽癌组织HSP27与β-catenin及Vimentin表达相关性研究

发布时间：2018-02-26 17:13

本文关键词： 鼻咽癌热休克蛋白 β-连接素波形蛋白　出处：《中华肿瘤防治杂志》2017年03期 　论文类型：期刊论文

【摘要】：目的研究发现,异常表达的热休克蛋白27(heat shock protein 27,HSP27)与多种肿瘤细胞的侵袭转移相关,但其在鼻咽癌(nasopharyngeal carcinoma,NPC)侵袭转移过程中的作用机制有待研究。因此,本实验通过检测NPC组织HSP27、β-连接素(β-catenin)及波形蛋白(Vimentin)的表达,探讨NPC组织中HSP27异常表达对Wnt/β-catenin信号通路及上皮间质转化(epithelial to mesenchymal transition,EMT)的影响。方法收集2013-01-01-2015-12-31海口市人民医院病理科病理确诊,为鼻咽未分化非角化性癌患者54例,免疫组化检测NPC组织中HSP27、β-catenin和Vimentin的表达。以维持编码氨基酸序列不变为前提,构建经密码子优化的HSP27cDNA质粒,瞬时转染人鼻咽癌CNE2细胞,荧光定量PCR(quantitative reverse transcription PCR,qRT-PCR)和蛋白质印迹法检测转染效果,并运用qRT-PCR和蛋白质印迹法检测转染后CNE2细胞β-catenin、Vimentin的转录和表达水平。结果HSP27高表达率为35.2%(19/54),HSP27中-低表达率为64.8%(35/54)。β-catenin在NPC组织中的胞质表达、膜表达和异质性表达率分别为42.6%(23/54)、27.8%(15/54)和29.6%(16/54);β-catenin异质性表达模式与胞质或胞膜模式病例的HSP27表达强度差异有统计学意义;Vimentin在NPC组织中的表达率为63.0%(34/54),与HSP27的表达强度存在低度相关,r=0.324,P=0.017。CNE2细胞在转染密码子优化的HSP27cDNA质粒后,Vimentin mRNA的下调差异有统计学意义,t=6.461,P=0.003;而β-catenin mRNA的下调差异无统计学意义,t=2.177,P=0.161;同时β-catenin和Vimentin的蛋白表达均呈下降趋势。结论 HSP27具有抑制NPC细胞Vimentin基因表达的功能,该作用可能与抑制β-catenin表达,从而阻抑Wnt/β-catenin信号通路活化有关。HSP27可能在调节NPC发生EMT和侵袭转移的过程起到重要作用。
[Abstract]:Objective to find that the abnormal expression of heat shock protein 27 (HSP27) is associated with the invasion and metastasis of various tumor cells, but its role in the invasion and metastasis of nasopharyngeal carcinoma (NPC) remains to be studied. By detecting the expression of HSP27, 尾 -catenin and vimentin Vimentin in NPC tissues, we investigated the effect of abnormal expression of HSP27 on Wnt/ 尾 -catenin signaling pathway and epithelial stromal transition to mesenchymal transition.Methods pathological diagnosis was collected from Haikou people's Hospital from January to 31, 2015-31, 2013-01-2015-12-31. The expression of HSP27, 尾 -catenin and Vimentin in NPC tissues was detected by immunohistochemistry in 54 patients with undifferentiated nonkeratinized nasopharyngeal carcinoma. In order to keep the coding amino acid sequence unchanged, a codon optimized HSP27cDNA plasmid was constructed and transiently transfected into CNE2 cells of nasopharyngeal carcinoma (NPC). Fluorescence quantitative PCR(quantitative reverse transcription PCR qRT-PCR and Western blot were used to detect the transfection effect. QRT-PCR and Western blot were used to detect the transcription and expression level of 尾 -catenin vimentin in transfected CNE2 cells. Results the high expression rate of HSP27 was 35.2g / 54 / 54 HSP27. The expression rate of 尾 -catenin in the cytoplasm of NPC was 64.8%. The HSP27 expression intensity of 尾 -catenin heterogenous expression pattern was significantly different from that of cytoplasm or cell membrane pattern in NPC tissues. The expression rate of vimentin in NPC tissues was 63.034 / 54, which was lower than that of HSP27. The down-regulation of Vimentin mRNA in codon optimized HSP27cDNA plasmid was significantly lower than that in 尾 -catenin mRNA, but the down-regulation of 尾 -catenin mRNA was not significant. The protein expression of 尾 -catenin and Vimentin showed a decreasing trend. Inhibition of Vimentin gene expression in NPC cells. This effect may be related to the inhibition of 尾 -catenin expression, thus inhibiting the activation of Wnt/ 尾 -catenin signaling pathway. HSP27 may play an important role in the regulation of EMT and invasion and metastasis in NPC.
【作者单位】：海口市人民医院病理科;海南医学院病理教研室;
【分类号】：R739.63

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