肝癌患者血清外泌体内miR-665的表达及临床意义

The expression of serum exosomal miR-665 and its correlation with clinical parameters in hepatocellular carcinoma

目的探讨肝癌患者血清来源的外泌体内miR-665与肝癌临床病理指标及预后的关系。方法采用试剂盒方法提取患者血清内的外泌体并用透射电镜观察验证。然后利用RT-PCR方法检测30例肝癌患者和10例体检者血清来源的外泌体内miR-665的表达水平,进行对照研究,探讨临床病理指标和预后的关系。结果血清提取的外泌体呈囊泡状,直径约30～150 nm。肝癌患者血清来源的外泌体miR-665较正常体检者表达水平明显升高,中位表达水平是对照组的8.3倍。外泌体miR-665高表达与患者肿瘤大小(>5 cm)(P=0.0042)、有包膜浸润(P=0.0197)以及临床TNM分期(Ⅲ～Ⅳ期)(P=0.0276)成明显相关,与患者年龄、性别、HBsAg以及AFP水平未见明显相关性(P> 0.05),外泌体miR-665高表达组患者生存期较低表达组更短(P=0.036)。结论肝癌患者血清中外泌体miR-665水平升高与肝癌进展相关,测定血清外泌体内miR-665的水平可能有助于肝癌的临床诊断和预后判断。

Objective To investigate the relationship of serum exosomal miR-665 expression and clinical pathological parameters and prognosis in hepatocellular carcinoma （HCC） patients. Methods Exosomes were isolated from patients serum by Exoquick TM Kit and proved by transmission elctron microscope （TEM）. Then, exosomal miR-665 levels were detected by RT-PCR method. The relationship between exosomal miR-665 expression and clinical pathological parameters and prognosis in HCC patients were analyzed. Results Exosomes were small membranous vesicles （50-150 nm）, exosomal miR-665 levels in HCC patients were signifcantly higher than that in healthy subjects （P〈0.05）. The exosomal miR-665 levels in HCC patients with larger tumor size （〉5 cm）, local invasion of tumor and advanced clinical stage （stage III/IV） were signifcantly higher than those in control normal subjects （P=0.0042, 0.0197, 0.0276, respectively）. The exosomal miR-665 levels were not signifcantly related to age, gender, HBsAg and AFP level in HCC patients （P〉0.05）. In addition, the survival time in HCC patients with exosomal miR-665 high expression （n=17） was signifcantly shorter than those patients with low expression （n=13） （P〈0.05）. Conclusion Increase of serum exosomal miR-665 levels in HCC patients is related to clinical stage and poor prognosis, which may be a potential biomarker of HCC diagnosis and prognosis.