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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。
Aims At the present study, we found that abnormal expression of heat shock protein 27 (HSP27) is involved in the invasion and metastasis of many kinds of tumor cells. However, its role in the invasion and metastasis of nasopharyngeal carcinoma (NPC) remains to be studied . Therefore, in this study, we investigated the expression of HSP27, β-catenin and Vimentin in NPC to investigate the effect of abnormal expression of HSP27 on the expression of Wnt / β-catenin signaling and epithelial epithelial to mesenchymal transition, EMT). Methods We collected 54 patients with undifferentiated non-keratinizing carcinoma of nasopharynx from 2013-01-01-2015-12-31 2013-01-01 2015-12-31 Pathology of Haikou People’s Hospital. Immunohistochemistry was used to detect the expression of HSP27, β-catenin and Vimentin expression. In order to maintain the coding amino acid sequence as a precondition, a codon-optimized HSP27 cDNA plasmid was constructed and transiently transfected into human nasopharyngeal carcinoma CNE2 cells. Quantitative reverse transcription PCR (qRT-PCR) and Western blotting were used to detect the transfection efficiency The transcription and expression of β-catenin and Vimentin in CNE2 cells were detected by qRT-PCR and Western blot. Results The high expression rate of HSP27 was 35.2% (19/54), while the low expression rate of HSP27 was 64.8% (35/54). The cytoplasmic, membrane and heterogeneous expression rates of β-catenin in NPC tissues were 42.6% (23/54), 27.8% (15/54) and 29.6% (16/54), respectively. The expressions of β-catenin There was significant difference in the expression intensity of HSP27 between the qualitative and cytoplasmic or cytomegalic cases; the expression rate of Vimentin in NPC was 63.0% (34/54), which had a low correlation with the expression intensity of HSP27, r = 0.324, P = 0.017. The down-regulation of Vimentin mRNA in CNE2 cells transfected with codon-optimized HSP27 cDNA plasmid was statistically significant (t = 6.461, P = 0.003), while there was no significant difference in down-regulation of β-catenin mRNA, t = 2.177, P = 0.161; At the same time, the protein expression of β-catenin and Vimentin showed a downward trend. Conclusion HSP27 can inhibit the expression of vimentin in NPC cells, which may be related to the inhibition of the expression of β-catenin and the activation of Wnt / β-catenin signaling pathway. HSP27 may play an important role in regulating the occurrence of EMT and invasion and metastasis in NPC.