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启动子甲基化及E-钙黏蛋白基因与蛋白表达水平在喉癌中的诊断价值

Diagnostic impact of promoter methylation and E-cadherin gene and protein expression levels in laryngeal carcinoma.

作者信息

Starska Katarzyna, Forma Ewa, Lewy-Trenda Iwona, Papież Paweł, Woś Jan, Bryś Magdalena

机构信息

Department of Otholaryngology and Laryngological Oncology, Medical University of Lodz, Poland.

Department of Cytobiochemistry, University of Lodz, Lodz, Poland.

出版信息

Contemp Oncol (Pozn). 2013;17(3):263-71. doi: 10.5114/wo.2013.35284. Epub 2013 Jun 28.

Abstract

AIM OF THE STUDY

Inactivation of the tumor suppressor E-cadherin (CDH1) and its decreased expression is an important occurrence during carcinogenesis. Nevertheless, the relationship of CDH1 expression and the promoter methylation with laryngeal cancer cell aggressiveness is still unclear. The purpose of this study was to elucidate the gene and protein E-cadherin expression and the DNA methylation levels and to describe the correlations with morphological features in squamous cell laryngeal cancer.

MATERIAL AND METHODS

The authors studied E-cadherin and the DNA methylation level in 86 cases to gain a further understanding of the clinicopathologic significance of analyzed parameters. The pathological evaluation included pTNM classification and the tumor front grading (TFG) criteria. Quantitative analysis of the amplified product in real time (qRT-PCR) for estimation of CDH1 mRNA was used. The methylation status was investigated by using methyl-specific polymerase chain reaction (MSP). The level of CDH1 protein expression by Western blot was determined.

RESULTS

Downregulation of E-cadherin was found to be related to promoter methylation (p < 0.001). In tumors with the highest invasiveness according to TFG criteria the lowest E-cadherin gene and protein level in the study group was observed (p = 0.046 and p = 0.0002, respectively). In SCLC with muscle and cartilage invasion and disperse infiltration the lowest CDH1 gene and protein expression was noted (p = 0.0003 and p = 0.003 for deep invasion, p = 0.033 and p = 0.003 for multifocal infiltration, respectively).

CONCLUSIONS

The current findings suggest an association of E-cadherin tumor expression with progression of laryngeal cancer. CDH1 gene level may be an auxiliary molecular marker for advanced cases of laryngeal carcinoma; however, further studies are necessary.

摘要

研究目的

肿瘤抑制因子E-钙黏蛋白(CDH1)的失活及其表达降低是致癌过程中的一个重要事件。然而,CDH1表达及启动子甲基化与喉癌细胞侵袭性之间的关系仍不清楚。本研究的目的是阐明E-钙黏蛋白的基因和蛋白表达、DNA甲基化水平,并描述其与喉鳞状细胞癌形态学特征的相关性。

材料与方法

作者研究了86例患者的E-钙黏蛋白和DNA甲基化水平,以进一步了解分析参数的临床病理意义。病理评估包括pTNM分类和肿瘤前沿分级(TFG)标准。采用实时定量分析扩增产物(qRT-PCR)来估计CDH1 mRNA。通过甲基特异性聚合酶链反应(MSP)研究甲基化状态。用蛋白质印迹法测定CDH1蛋白表达水平。

结果

发现E-钙黏蛋白的下调与启动子甲基化有关(p < 0.001)。在根据TFG标准具有最高侵袭性的肿瘤中,研究组观察到最低的E-钙黏蛋白基因和蛋白水平(分别为p = 0.046和p = 0.0002)。在伴有肌肉和软骨侵犯及弥漫性浸润的小细胞肺癌中,观察到最低的CDH1基因和蛋白表达(深部侵犯时p = 0.0003和p = 0.003,多灶性浸润时p = 0.033和p = 0.003)。

结论

目前的研究结果表明E-钙黏蛋白在肿瘤中的表达与喉癌进展有关。CDH1基因水平可能是晚期喉癌的辅助分子标志物;然而,还需要进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db18/3934075/717ff3b0406d/WO-17-20860-g001.jpg

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