摘要
目的研究DACH1在肺癌组织及配对癌旁组织中的表达情况,以及对人肺腺癌细胞增殖、侵袭和凋亡的作用。方法收集46例经病理学诊断为肺癌患者的组织标本,选取肿瘤组织并以配对癌旁组织为对照,应用实时荧光定量PCR检测肿瘤组织与配对癌旁组织标本中DACH1mRNA表达情况;采用免疫组织化学法检测两组标本中DACH1蛋白的表达情况。使用siRNA干扰技术抑制人肺腺癌A549细胞系中DACH1的表达,采用MTT法观察其对肿瘤细胞增殖的影响,Transwell小室法观察肿瘤细胞侵袭的变化,采用流式细胞术检测对细胞系诱导凋亡的作用。结果实时荧光定量PCR测得DACH1mRNA在肺癌组织中表达下降,免疫组织化学检测结果显示肺癌组织中DACH1蛋白表达明显降低,差异有统计学意义。siRNA干扰A549细胞DACH1的表达后,MTT法显示,si-DACH1组细胞增殖能力明显增强;Transwell小室法显示si-DACH1组细胞侵袭能力明显增强,流式细胞术显示,si-DACH1组细胞自发凋亡率明显降低。结论 DACH1在肺腺癌组织中表达减少,在肺腺癌中起到抑癌基因的作用,并有望成为肺癌治疗的新靶点。
Objective To examine the expression of DACH1 mRNA and protein in lung tumor tissues and their adjacent normal tissues and the role of DACH1 in the proliferation,invasion and apoptosis of lung adenocarcinoma cells.Methods The expression of DACH1 mRNA was detected by real-time fluorescent quantitative PCR in lung adenocarcinomas（n=46）and the matched adjacent normal tissues（n=46）.Immunohistochemistry was used to detect the expression of DACH1 protein.Small-interfering（si）RNA was used to inhibit the expression of DACH1 in A549cells.After down-regulation of DACH1 with siRNA,the proliferation of A549 cells was evaluated by MTT assay,the invasive ability of cells by using Transwell chamber assay,and the cell apoptosis by flow cytometry.ResultsReal-time fluorescent quantitative PCR and immunohistochemistry showed that the levels of DACH1 mRNA and protein were significantly decreased in lung adenocarcinomas as compared with those in normal tissues.Down-regulation of DACH1 by siRNA resulted in a significant increase in the proliferation and invasion of A549 cells.Flow cytometry revealed that the spontaneous apoptosis was significantly decreased in A549 cells with down-regulation of DACH1.Conclusion The expression of DACH1 was profoundly decreased in lung adenocarcinomas,indicating that DACH1 may serve as a tumor suppressor gene in lung adenocarcinomas and it is expected to become a new therapeutic target for lung cancer.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2016年第4期361-365,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
湖北省自然科学基金资助项目(No.2013CF088)
关键词
肺腺癌
DACH1
增殖
侵袭
凋亡
lung adenocarcinoma
DACH1
proliferation
invasion
apoptosis
