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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 investigate the proliferation and apoptosis-inducing effects of imiquimod on human skin squamous cell carcinoma SCL-1 cell line in vitro. Methods MTT was used to analyze the proliferation of SCL-1 cells in human skin squamous cell carcinoma. The morphological changes were observed under inverted microscope. The apoptosis of SCL-1 cells was detected by flow cytometry with Annexin V / PI. Fluorescence staining of acridine orange The morphological changes of late apoptotic cells were detected. The extent of apoptosis was detected by the cell death detection kit. The expressions of Fas and bcl-2 were detected by immunohistochemistry and RT-PCR. Results Imiquimod significantly inhibited the proliferation of human skin squamous cell carcinoma SCL-1 cells and resulted in morphological changes. The apoptosis rate of SCL-1 cells was the highest at 24 h after 0.150 mg / mL imiquimod treatment, reaching 47.23%. After 48 h, typical apoptosis bodies were observed. The degree of apoptosis in SCL-1 cells was increased with the increase of imiquimod concentration and the action time prolonged. The effect of imiquimod could up-regulate Fas and down-regulate bcl-2 gene expression. Conclusion Imiquimod can significantly inhibit the growth of SCL-1 tumor cells and induce apoptosis.