miR-25对食管癌EC109细胞侵袭和迁移能力的 影响及临床意义

目的 探索 miR-25 对食管癌侵袭和迁移能力的影响以及作为食管癌诊断生物标志物的潜力。方法 （1）荧光定量PCR（qPCR）检测54例早期食管癌患者癌组织和癌旁组织中miR-25表达水平。（2）人食管癌细胞EC109 分为miR-25 mimic组、NC mimic组、miR-25 inhibitor组和NC inhibitor组。转染相应序列后，qPCR检测miR-25转染 效率。Transwell实验检测过表达或敲低miR-25对EC109细胞侵袭和迁移的影响。Targetscan数据库预测miR-25的 靶基因，选定靶基因盐诱导激酶1（SIK1）基因。Western blot 和双荧光素酶报告实验鉴定miR-25与SIK1的靶向关 系。（3）EC109 细胞分为 pcDNA 3.1 组、SIK1 过表达组和 miR-25+SIK1 过表达组。Transwell 实验检测 miR-25 靶向 SIK1对EC109细胞侵袭和迁移能力的影响。（4）提取食管癌患者（54例）和健康对照者（54例）的血浆外泌体，比较2 组血浆外泌体中miR-25的相对表达量，分析食管癌患者癌组织和血浆外泌体中miR-25的相关性。结果 （1）食管 癌组织中 miR-25 相对表达水平高于癌旁组织。（2）过表达 miR-25 后，EC109 细胞侵袭和迁移能力增强，而敲低 miR-25 表达后，细胞侵袭和迁移能力下降。Western blot 结果显示，过表达 miR-25 后，SIK1 蛋白表达下降；反之， 敲低miR-25后SIK1蛋白表达升高。（3）Transwell实验显示，与pcDNA 3.1组相比，SIK1过表达组EC109细胞侵袭和 迁移的细胞数量减少，而miR-25和SIK1联合作用后，EC109侵袭和迁移的细胞数量较SIK1过表达组增多。（4）食管 癌血浆外泌体样本中miR-25的表达量高于健康对照，且miR-25在食管癌组织中的表达与血浆外泌体中的相对表达 量呈正相关。结论 miR-25可能通过靶向调控SIK1来促进食管癌细胞的侵袭和迁移，且miR-25有作为食管癌早 期诊断生物标志物的潜力。

Effects of miR-25 on invasion and migration of esophageal carcinoma EC109 cells and its clinical significance

Objective To explore the effect of miR-25 on invasion and migration of esophageal cancer and its potential ability as a biomarker for the diagnosis of esophageal cancer. Methods (1) The expression levels of miR-25 in cancer tissues and adjacent tissues of 54 patients with esophageal cancer were detected by fluorescence quantitative PCR (qPCR). (2) Human esophageal cancer cells EC109 were divided into the miR-25 mimic group, the NC mimic group, the miR-25 inhibitor group and the NC inhibitor group. After transfection of the corresponding sequences, miR-25 overexpression and interference efficiency were detected by qPCR. Transwell assay was used to detect the effect of overexpression or silencing of miR-25 on the invasion and migration of EC109 cells. Targetscan database predicted the target gene of miR-25 and selected the target gene salt-induced kinase 1 (SIK1). Western blot assay and double luciferase report experiment identified the targeting relationship between silencing and overexpression of miR-25 and SIK1. (3) EC109 cells were divided into the pcDNA 3.1 group, the SIK1 overexpression group and the miR-25 + SIK1 overexpression group. Transwell assay was used to detect the effect of miR-25 targeting SIK1 on the invasion and migration of EC109 cells. (4) The plasma exosomes of patients with esophageal cancer (54 cases) and healthy controls (54 cases) were extracted, the relative expression of miR-25 in plasma exosomes was compared between the two groups. The correlation between miR-25 in tissues and plasma exosomes of patients with esophageal cancer was analyzed. Results (1) The relative expression level of miR-25 was higher in esophageal carcinoma tissues than that in adjacent tissues. (2) After overexpression of miR-25, the invasion and migration ability of EC109 cells increased, while after silencing miR-25 expression, the invasion and migration ability of EC109 cells decreased. The Targetscan database predicted that SIK1 was the target gene of miR-25. Western blot assay showed that the expression of SIK1 protein decreased after miR-25 overexpression. On the contrary, the protein expression of SIK1 increased after knocking down the expression of miR-25. (3) Transwell experiment showed that compared with the pcDNA 3.1 group, the number of invasion and migration of EC109 cells decreased in SIK1 overexpression group, while the number of invasion and migration of EC109 cells increased in the miR-25 and SIK1 overexpression group compared with the SIK1 overexpression group. (4) The expression of miR-25 in plasma exosomes of esophageal cancer was higher than that in healthy controls, and the expression of miR-25 in esophageal cancer was positively correlated with that in plasma exosomes. Conclusion miR-25 may promote the invasion and migration of esophageal cancer through targeting the regulation of SIK1, and miR-25 has the potential as a biomarker for the early detection of esophageal cancer.