胃癌患者血清miR-181d与PDCD4的表达及意义

安娟; 潘元明; 康倩; 王昕; 孙伟; 晏宇鹏; 郑伟

doi:10.3971/j.issn.1000-8578.2019.18.0486

胃癌患者血清miR-181d与PDCD4的表达及意义

Expression and Clinical Significance of miR-181d in Serum and PDCD4 in Gastric Cancer Patients

摘要

摘要:
目的分析胃癌患者血清miR-181d和PDCD4的表达及调控机制。
方法利用Real-time PCR检测6株胃黏膜细胞系中miR-181d和PDCD4的表达，并通过Western blot检测PDCD4的表达；采用Real-time PCR检测58例胃癌患者和50例健康体检者血清miR-181d的表达，并通过ELISA方法检测PDCD4的表达；采用化学法合成干扰/过表达miR-181d及构建PDCD4 3' -UTR荧光报告素载体检测miR-181d对PDCD4的调控。
结果 Real-time PCR结果显示miR-181d和PDCD4在胃癌细胞系中存在差异表达；miR-181d在胃癌患者血清中的表达高于健康体检者（P < 0.001），PDCD4表达则降低（P < 0.05）, 通过临床相关性分析表明，胃癌血清miR-181d与PDCD4呈显著负相关（R²=-0.44），并在健康体检者中miR-181d与PDCD4蛋白呈负相关（R²=-0.426）；结合临床资料分析，血清miR-181d的表达与吸烟喝酒史呈正相关（P=0.015, P=0.034），而靶基因PDCD4仅与饮酒史相关（P < 0.001）；ROC分析显示血清miR-181d与PDCD4用于体外诊断区分肿瘤和健康体检者具有良好的特异性与敏感度。过表达miR-181d于AGS细胞后PDCD4表达下降；反义miR-181d转染BGC823细胞后PDCD4表达增加。双荧光报告系统发现miR-181d可作用于PDCD4 3' -UTR区。
结论在胃癌患者血清中miR-181d的表达较高，而PDCD4的表达较低，并且miR-181d可能是通过作用于PDCD4 3' -UTR区发挥作用。

Abstract:
Objective To investigate miR-181d and PDCD4 expression in the serum of gastric cancer(GC) patients and regulatory mechanism.
Methods The expression of miR-181d and PDCD4 were detected in 6 gastric cell lines using Real-time PCR and Western blot. Real-time PCR was used to detect miR-181d expression in serum of 58 GC patients and 50 healthy volunteers, and the protein level of PDCD4 was detected by ELISA assay. Furthermore, the experiment of siRNA or mimics was used to evaluate the role of miR-181d on the regulation of PDCD4 in GC cells, and we constructed the PDCD4 3'-UTR luciferase report vector to analyze the association between miR-181d and PDCD4.
Results miR-181d and PDCD4 were differentially expressed in GC cell lines; miR-181d in the serum of GC patients was higher than that in normal group (P < 0.001), while the expression of PDCD4 protein in GC patients was decreased (P < 0.05). It was found that there was a significant inverse correlation between miR-181d and PDCD4 expression in GC patients (R²=-0.44) and also in normal group(R²=-0.426). ROC curve analysis showed that miR-181d and PDCD4 had good specificity and sensitivity for the diagnosis of GC and normal population in vitro. Moreover, miR-181d expression was associated with smoking and drinking history, respectively (P=0.015, P=0.034). The expression of PDCD4 was only associated with drinking history(P < 0.001). Overexpression of miR-181d in AGS cells decreased the expression of PDCD4, and the expression of PDCD4 was increased after transfection with antisense miR-181d in BGC823 cells. Luciferase reporter system results suggested that miR-181d could bind to the 3'-UTR region of PDCD4.
Conclusion Elevated miR-181d expression and downregulated PDCD4 expression are detected in the serum of GC patients; miR-181d may mediate PDCD4 expression in the development of gastric cancer.

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