STK15过表达对人食管癌细胞株KYSE150生长的影响  

Effects of STK15 overexpression on growth of human esophageal carcinoma cell line KYSE150

作　　者：王晓霞[1] 李晓钟[2] 路娜[1] 赵艳[1] 王康[1] 裴毅[3] 

机构地区：[1]山西医科大学生物化学与分子生物学教研室,山西太原030001 [2]山西省人民医院急诊科,山西太原030012 [3]山西医学科学院,山西大医院老年肿瘤科,山西太原030032

出　　处：《中国病理生理杂志》2013年第11期1957-1961,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30973401);山西省自然科学基金资助项目(No.2009011052)

摘　　要：目的:探讨丝氨酸/苏氨酸激酶15(serine/threonine kinase 15,STK15)过表达对人食管癌细胞株KYSE150生长的影响。方法:采用脂质体法将pEGFP-C1-STK15真核表达载体转染人食管鳞癌细胞株KYSE150,构建STK15过表达的稳定细胞系(GFP-STK15),通过荧光显微镜和Western blotting检测STK15的表达。采用四甲基偶氮唑蓝(MTT)法观察STK15过表达对体外肿瘤细胞生长的影响。通过流式细胞术分析STK15过表达对细胞周期和凋亡的影响。采用裸鼠致瘤实验观察STK15过表达对体内肿瘤细胞生长的影响。结果:pEGFP-C1-STK15质粒转染细胞后筛选得到稳定克隆,荧光显微镜显示GFP-STK15融合蛋白定位于细胞的中心体以及纺锤体,Western blotting可检测到GFP-STK15融合蛋白的表达。与对照细胞相比,GFP-STK15细胞系的体外生长能力明显增加;G0/G1期细胞百分率降低,细胞凋亡率减少;GFP-STK15细胞系接种组裸鼠肿瘤明显增大,差异均具有统计学意义(P<0.01)。结论:STK15过表达能促进人食管癌KYSE150细胞的生长,提示STK15将可能成为治疗食管癌的新靶点。AIM： To investigate the effects of serine/threonine kinase 15 （STK15） overexpression on the growth of human esophageal squamous-cell carcinoma （ESCC） cell line KYSE150. METHODS： Recombinant pEGFP- C1-STK15 expression vector was transfected into KYSE150 cells using LipofectamineTM 2000 and the expression of STK15 was detected by fluorescence microscopy and Western blotting. The proliferation of the cells in vitro was measured by MTT assay. The cell cycle distribution and apoptosis were detected by flow cytometry. The proliferation of the cells in vivo was measured by tumorigenicity experiment in nude mice. RESULTS： After recombinant pEGFP-C1-STK15 expression vector was stably transfected into KYSE150 cells, GFP-STK15 fusion protein localized to centrosome and spindle. The STKIS- overexpressing colonies were further confirmed by Western blotting. MTr assay showed that the proliferation of the cells in STK15 overexpression group was increased compared with control group （P 〈0.01 ）. Flow cytometry analysis showed that the percentage of the cells in G0/G1 phase and the cell apoptosis in STK15 overexpression group were decreased compared with control group （P 〈 0.01 ）. The tumorigenicity experiment in nude mice showed that the proliferation of the cells in STK15 overexpression group was increased compared with control group （P 〈0.01 ）. CONCLUSION： Overexpression of STK15 in human ESCC KYSE150 cells promotes the cell growth in vitro and in vivo, indicating that STK15 may serve as a novel theraveutic target for esovhazeal carcinoma.

关 键 词：丝氨酸 苏氨酸激酶15 食管肿瘤 细胞增殖 

分 类 号：R735.1[医药卫生—肿瘤]