摘要
目的 探讨咪喹莫特对人皮肤鳞状细胞癌SCL—1细胞增殖及端粒酶活性的影响。方法使用噻唑蓝(MTT)法检测药物处理后细胞生长抑制情况,细胞克隆形成试验检测药物处理后细胞增殖能力变化,倒置显微镜观察药物作用前后细胞形态改变;端粒酶重复序列扩增-酶联免疫吸附法(TRAP—ELISA)检测细胞端粒酶活性变化;细胞免疫组化和实时定量RT—PCR技术检测端粒酶活性相关c—Mvc基因表达情况。结果咪喹莫特能明显抑制SCL—1细胞增殖,并存在浓度依赖性。培养48h时,0,0.05,0.10,0.15和0.20g/L咪喹莫特组细胞增殖抑制率分别为0,31.3%±3.19%,44.7%±4.21%,49.2%±3.71%和71.6%±3.24%。咪喹莫特作用SCL—1细胞48h,端粒酶活性随药物浓度增高呈显著的递减关系。48h时,0.05和0.15g/L咪喹莫特组c—Myc mRNA和蛋白表达均低于空白对照组(P〈0.05)。结论 咪喹莫特可明显抑制SCL—1细胞的增殖,降低c—Myc基因表达,下调SCL—1肿瘤细胞端粒酶活性,可能是其抗癌作用的机制之一。
Objective To investigate the effect of imiquimod on the proliferation and telomerase activity of human skin squamous cell carcinoma cell line SCL-1. Methods Various concentrations of imiquimod was used to treat cultured SCL-1 cells in log phase. After 48 hours of culture, MTT assay and clonogenic assay were used to assess the proliferation of cells, the morphological alterations were observed by transmission electron microscopy, and telomerase activity was detected with telomeric repeat amplification protocol enzyme-linked immunoadsorbent assay (TRAP-ELISA). The protein and mRNA expressions of c-Myc were analyzed by immunohistochemistry and real time quantitative RT-PCR, respectively. Results The 48-hour treatment with imiquimod at the concentrations of 0.05, 0.10, 0.15 and 0.20 g/L down-regulated the proliferation of SCL-1 cells by 31.3% ± 3.19%, 44.7% ± 4.21%, 49.2%±3.71% and 71.6% ± 3.24%, respectively, and telomerase activity of SCL-1 cells was also inhibited by imiquimod; both effects were concentration-dependant. A reduction in mRNA and protein expressions of c-Myc was observed in SCL-1 cells after the treatment with imiquimod at 0.05 and 0.15 g/L, respectively (both P 〈 0.05). Conclusion Imiquimod can effectively inhibit the proliferation and telomerase activity of SCL-1 cells, decrease the expression of c-Myc gene, which may be associated with the mechanism of antitumor effect of imiquimod.
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2008年第3期176-179,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: