miR-196b-5p在食管鳞癌癌组织中的表达及作用机制

Expression of miR-196b-5p in esophageal squamous cell carcinoma and its clinical significance

目的探讨miR-196b-5p在食管鳞癌癌组织中的表达情况,评估miR-196b-5p作为食管鳞癌潜在生物标志物的可能性,并进一步分析其作用机制。方法收集2015至2019年浙江省台州医院收治的病理学检查确诊为食管鳞癌的24例患者的癌组织及相应癌旁(距离肿瘤≥2 cm)正常组织标本。采用qRT-PCR法检测miR-196b-5p表达水平。ROC曲线评估miR-196b-5p诊断食管鳞癌的效能,并收集癌症基因组图谱(TCGA)和基因表达汇编(GEO)数据进行验证。进一步比较不同临床特征食管鳞癌患者癌组织中miR-196b-5p表达水平。基于TCGA数据库,通过京都基因与基因组百科全书(KEGG)富集方法初步分析miR-196b-5p的相关信号通路。结果qRT-PCR检测结果、TCGA数据库和GEO数据库中食管鳞癌患者癌组织miR-196b-5p表达水平均明显高于癌旁正常组织(均P<0.01)。miR-196b-5p水平诊断食管鳞癌的灵敏度为0.79,特异度为0.92,最佳截断值为1.542,AUC为0.891(95%CI:0.767~0.962)(P<0.01);根据TCGA数据计算,灵敏度为0.86,特异度为0.85,最佳截断值为5.310,AUC为0.832(95%CI:0.748~0.897)(P<0.01);根据GEO数据计算,灵敏度为0.89,特异度为0.96,最佳截断值为0.012,AUC为0.946(95%CI:0.900~0.975)(P<0.01),T3~4期食管鳞癌患者癌组织中miR-196b-5p表达水平高于T1~2期食管鳞癌患者(P<0.05)。食管鳞癌癌组织miR-196b-5p低表达(≤1.898)患者3年生存率为55.3%,明显高于miR-196b-5p高表达(>1.898)患者的0.0%(P<0.05)。KEGG富集结果提示差异基因显著富集于有丝分裂原活化蛋白激酶(MAPK)、Wnt、磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)等多条通路(均P<0.05)。结论食管鳞癌癌组织中miR-196b-5p表达水平明显高于癌旁正常组织,或可作为食管鳞癌筛选的潜在生物标志物。miR-196b-5p可能通过参与调控多条信号通路,在食管鳞癌中起致癌作用。

Objective To study the expression of miR-196b-5p in esophageal squamous cell carcinoma tissues and its clinical significance.Methods Twenty-four cancer specimens and 24 corresponding paracancerous normal tissues samples were taken from patients with esophageal squamous cell carcinoma from 2015 to 2019.The expression level of miR-196b-5p in cancer tissues and adjacent normal tissues was detected by qRT-PCR.ROC curve was used to analyze the sensitivity and specificity of miR-196b-5p for diagnosis of esophageal cancer.TCGA and GEO database data were collected for validation.The association of miR-196b-5p expression with clinicopathological features of cancer were analyzed.Based on TCGA database,the signal pathway of miR-196b-5p was analyzed by KEGG enrichment.Results The qRT-PCR testing and the TCGA and GEO data analysis showed that the expression level of miR-196b-5p in esophageal squamous cell carcinoma was significantly higher than that in normal tissues(all P<0.01).The sensitivity of miR-196b-5p in the diagnosis of esophageal squamous cell carcinoma was 0.79,specificity was 0.92,best cut-off value was 1.542,AUC was 0.891(95%CI:0.767-0.962)(P<0.01).According to TCGA data,the sensitivity was 0.86,specificity was 0.85,best cut-off value was 5.310,AUC was 0.832(95%CI:0.748-0.897)(P<0.01).According to GEO data,the sensitivity was 0.89,specificity was 0.96,best cut-off value was 0.012,AUC was 0.946(95%CI:0.900-0.975)(P<0.01).The expression level of miR-196b-5p in cancer tissues of patients with stage T3-4 was higher than that with stage T1-2(P<0.05).The 3-year survival rate of patients with low expression of miR-196b-5p was significantly higher than that with low expression of miR-196b-5p(P<0.05).KEGG enrichment results showed that the differential genes were significantly enriched in MAPK,Wnt,PI3K/Akt and other pathways(all P<0.05).Conclusion The expression level of miR-196b-5p is up-regulated in esophageal squamous cell carcinoma tissue,which may be used as a potential biomarker for screening esophageal squamous cell carcinoma.miR-196b-5p may play a carcinogenic role in esophageal squamous cell carcinoma by regulating multiple signaling pathways.