微小RNA-920靶向调控Yes相关蛋白1增强食管癌的化疗敏感

MicroRNA -920 enhances chemosensitivity in esophageal cancer by targeting yes -associated pro- tein 1

目的探讨Yes相关蛋白1（YAP1）在食管癌组织中的表达及其与化疗耐受的关系，以及微小RNA（miRNA，miR）-920在YAP1表达调控中的作用。方法免疫组织化学检测YAP1在30例食管癌组织及其正常组织中的表达，小干扰RNA（siRNA）干扰技术结合聚合酶链反应（PCR）及四唑氮化合物（MTS）实验在食管癌细胞株中检测YAPl表达与化疗敏感性的关系。流式细胞仪分选食管癌肿瘤干细胞群，结合PCR及WesternNot方法检测YAPl在肿瘤干细胞的表达。siRNA干扰技术结合干细胞微球体培养检测YAP1在肿瘤干细胞活性维持中的作用。运用生物信息学软件预测靶向YAP1的miRNAs，同时利用miRNA定量PCR确定候选miRNA的表达差异。转染miRNA模拟物结合Westernblot实验验证候选miRNA对YAP1靶向调节作用。最后利用四唑氮化合物（MTS）方法检测miR-920在食管癌化疗中的作用。结果YAP1在食管癌癌组织中表达（5．300±0．559）明显高于食管正常组织（2．200±0．327，P=0．001），干扰YAPl可增强食管癌化疗敏感性（DDP组：P=0.003，PTX组：P=0．000），YAP1在食管癌肿瘤干细胞群中表达（5．667±0．636）明显高于非干细胞群（1．267±0．176，P=0．002），miR-920在食管癌中表达降低与YAP1表达呈负相关。结论miR-920可靶向抑制YAP1表达，增强食管癌的化疗敏感性。

Objective To study the expression of protein yes- associated protein 1 （YAP1） in esophageal carcinoma and its relationship with chemotherapy tolerance and the role of microRNA （ miRNA, miR） - 920 in the regulation of YAP1 expression. Methods Immunohistochemical technique was used to detect the expression of YAP1 in 30 cases of esophageal cancer tissues and paired normal tissues. Small in- terfering RNA （siRNA） interference technology combined with real - time quantitative polymerase chain reaction （ Real - time PCR） and 3 - （ 4,5 - Dimethylthiazol - 2 - yl） - 5 - （ 3 - carboxymethoxyphenyl） - 2 - （4 - sulfophenyl） - 2H - tetrazolium （MTS） experiment was used to detect the expression of YAP1 and its correlation with chemosensitivity in esophageal cancer cell lines. The esophageal cancer stem ceils were sorted by flow cytometry, and the expression of YAP1 was examined by Real - time PCR and Western blot- ting. The siRNA interference technology in combination with stem cell microspheres culture method was used to test the role of YAP1 in maintaining activity of tumor stem celis. Bioinformatics software was used to predict miRNAs targeting YAP1, and miRNA quantitative FCR was used to determine the expression difference of candidate miRNA. Transfection of miRNA analog combined with Western blotting was carried out to verify the targeting regulatory effects of the candidate miRNA on YAP1. Finally, MTS was used to detect the role of miR - 920 in the chemotherapy of esophageal carcinoma. Results The expression of YAP1 in 30 cases of esophageal carcinoma tissues （5. 300 ± 0. 559） was obviously higher than normal tissues （2. 200 ±0. 327, P = 0. 001 ）, and interfering with YAP1 could increase the chemosensitivity of esophageal carcinoma （ DDP group, P = 0. 003, PTX group, P = 0. 000）. Moreover, the expression of YAP1 in the tumor stem cell populations （SP） of esophageal cancer （5.667±0. 636） was obviously higher than non - SP cell （ 1. 267 ± 0. 176, P = 0. 002）. In addition, the expression of miR - 920 in esophageal cancer was negatively correlated with the expression of YAP1. Conclusion MiR - 920 can inhibit the expression of YAP1 and increase the chemosensitivity of esophageal carcinoma.