咪喹莫特对皮肤鳞状细胞癌SCL-1细胞株增生及凋亡的影响  

Effect of imiquimod on the proliferation and apoptosis of human skin squamous cell carcinoma cell line SCL-1

作　　者：陈江[1] 何春涤[1] 王雅坤[1] 王珍[1] 金鑫[1] 肖汀[1] 姜奕[1] 陈洪铎[1] 

机构地区：[1]中国医科大学附属第一医院皮肤科,沈阳110001

出　　处：《中华皮肤科杂志》2007年第9期564-567,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金(30271198);教育部留学回国人员科研启动基金[2002]247号

摘　　要：目的探讨咪喹莫特对体外培养人皮肤鳞状细胞癌SCL-1细胞株的增生抑制及凋亡诱导作用。方法采用MTT法分析咪喹莫特对人皮肤鳞状细胞癌SCL-1细胞的增生,倒置显微镜观察细胞形态学,Annexin V/PI双染流式细胞仪检测细胞凋亡,吖啶橙荧光染色检测细胞凋亡晚期形态改变,细胞死亡检测试剂盒检测细胞凋亡的程度,细胞免疫组化和RT-PCR检测凋亡相关基因Fas、bcl-2的表达。结果咪喹莫特可明显抑制人皮肤鳞状细胞癌SCL-1细胞的增殖,并可导致其形态学改变。经0.150 mg/mL咪喹莫特作用后24 h,SCL-1细胞早期凋亡率最大,达47.23%;作用48 h,可见典型凋亡小体。SCL-1细胞凋亡程度随咪喹莫特浓度增高、作用时间延长呈递增关系,经咪喹莫特作用后可上调Fas和下调bcl-2基因表达。结论咪喹莫特可明显抑制SCL-1肿瘤细胞生长,诱导细胞凋亡。Objective To study the effect of imiquimod on SCL-1 cell proliferation and apoptosis. Methods SCL-1 cells were cultured in 96-well microplates and treated with various concentrations of imiquimod for different durations. The proliferation of SCL- 1 cells was measured using MTT assay, the morphological alterations observed by an inverted microscope, the early apoptosis of cells evaluated by flow cytometry with Annexin V and PI, and the morphological changes at late apoptosis observed by acridine orange staining. Cell death detection kit was utilized to count the apoptosis cells, RT-PCR and immunohistochemical method to detect the expressions of apoptosis-associated genes, Fas and bcl-2. Results Imiquimod could significantly inhibit the growth, and cause morphological changes, of SCL-1 cells. The early apoptosis rate was 47.23% in SCL-1 cells treated with 0.15 mg/ml imiquimod for 24 h, and was the highest among the cells treated with various durations of imiquimod. Typically apoptotic bodies were observed in the SCL-1 cells treated with imiquimod for 48 h. Imiquimod induced the apoptosis of SCL- 1 cells in a time- and dose- dependent fashion. Moreover, the expression of Fas was elevated and that of bcl-2 was decreased by the treatment of imiquimod. Conclusion Imiquimod could markedly inhibit the proliferation of SCL-1 cells and induce the apoptosis of them.

关 键 词：癌 鳞状细胞 细胞系 肿瘤 咪喹莫特 SCL—1细胞 细胞增殖 细胞凋亡 

分 类 号：R739.5[医药卫生—肿瘤]