miR31对人皮肤鳞状细胞癌生长的影响及作用机制的研究.pdfVIP

中国肿瘤临床2014年第41卷第9期 ChinJClinOncol2014,Vol.41,No.9 555 miR-31对人皮肤鳞状细胞癌生长的影响及作用 机制的研究 陈 凯 常东方 段绍坤 李渝凉 探讨miR-31对皮肤鳞状细胞癌（鳞癌）生长的影响及作用机制。 在鳞癌细胞中转染miR-31的2-羟基甲 摘要 目的： 方法： 基化的反义寡核苷酸（antisenseoligonueleotide，ASO），运用平板克隆形成和体外成瘤实验检测miR-31对鳞癌生长的影响。运用 Westernblot印迹及GFP报告基因实验验证miR-31的靶基因。在鳞癌细胞中转染靶基因的siRNA，检测其对细胞生长的影响。 最后，运用实时定量PCR以及免疫组织化学检测miR-31和靶基因在鳞癌组织中的水平。结果：转染miR-31反义寡核苷酸 （miR-31ASO）可以抑制克隆数目及体内裸鼠肿瘤体积（P0.05）。LATS2（largetumorsuppressorhomolog2）是miR-31的直接靶基 因。抑制LATS2表达后，鳞癌细胞克隆数目增加（P0.05）。miR-31在鳞癌中高表达，与LATS2的表达呈负相关。免疫组织化学 结果也显示LATS2在鳞癌组织中低表达。 miR-31通过负调控LATS2抑制鳞癌的生长。因此，miR-31具有作为鳞癌分子治 结论： 疗靶标的潜能。 关键词 微小RNA 鳞状细胞癌 荧光定量PCR doi:10.3969/j.issn.1000-8179 Roles and regulation mechanism of miR-31 in human cutaneous squamous cell carcinoma growth Kai CHEN, Dongfang CHANG, Shaokun DUAN, Yuliang LI Correspondence to: Kai CHEN; E-mail: chenkai01213@163.com Department of Burns and Plastic Surgery, the Affiliated Yongchuan Hospital of Chongqing Medical University, Chongqing 402160, China. Abstract Objective: To investigate the roles and regulation mechanism of miR-31 in human cutaneous squamous cell carcino- ma (cSCC) growth. Methods: cSCC cells were transfected with the antisense oligonucleotide (ASO) of miR-31, and the cSCC growth was tested by colony formation and in vivo tumor formation assays. The target gene of miR-31 was validated by Western blot and green fluorescent protein (GFP) reporter assay. The cells were then transfected with the siRNA of the target gene, and the effect of the target gene on cell growth was preformed by colony formation assay. Finally, real-time PCR and immunohistochemistry were us