黄芩素抑制皮肤鳞癌细胞株A431增殖的体外研究

Investigation of inhibitory effects of baicalein on proliferation of cutaneous squamous carcinoma A431 cell line in vitro

目的研究黄芩素(BAI)对鳞状细胞癌A431细胞的增殖、集落形成及细胞周期的影响,探索BAI抗肿瘤的作用机制。方法以体外培养的人表皮鳞癌A431细胞株为研究对象,MTT法观察BAI对A431细胞的生长抑制作用,光镜和电镜观察细胞形态,集落形成实验观察BAI对瘤细胞的生长抑制,流式细胞仪检测细胞周期中各期细胞比例和凋亡率。结果 MTT法显示,随着BAI作用时间延长及剂量增加对A431细胞增殖的抑制作用增强,有明显的时间和剂量依赖性,与对照组比较差异有显著性(P<0.01);光镜显示,随黄芩素浓度的增大细胞生长速度明显受到抑制,形态变圆;电镜显示,A431细胞伪足明显缩短,数量减少;流式细胞仪检测显示,随BAI浓度及作用时间的延长,细胞周期被明显阻断在S期,凋亡率增高,与对照组比较差异有显著性(P<0.05)。结论①BAI处理48h后,抑制A431细胞增殖而无细胞毒性的最适剂量范围为10～30μM;②BAI能明显地抑制皮肤鳞癌A431细胞增殖、细胞周期的进程、瘤细胞集落及伪足形成。

【Objective】 To investigate the effect of baicalein on proliferation,cell-cycle,pseudopodia and colony formation on epithelial-squamous-cell carcinoma-A431 cell-line and to find out the optimal inhibitory concentration of baicalein in A431 cell-line without cytotoxicity.【Methods】 MTT assay was applied to survey proliferation-inhibitory effect of baicalein on A431 cell,normal-optic-microscope was employed to observe cell growth and their Morphologic change,scanning electron microscope was used to view the pseudopod formation of A431 cell-line,colony formation assay was applied to observe the inhibitory-effect of baicalein on tumor formation of A431 neoplastic cell line,cell cycle progression analysis was performed by flow cytometry to indicate cell percentage of each stage and apoptosis ratio in A431 cell line treated by baicalein.【Results】 Methylthiazolyl tetrazolium（MTT） analysis showed that the depressant effect of baicalein on proliferation of A431 cell line was enhanced with its action time-extending and dose-adding,it indicated obviously that time and dose were dependent,and it is significant to compare with control group（P 0.01）.With baicalein concentration-increasing,the growth velocity of A431 cells was obviously pressed down viewing under a optic-microscope,morphous became round and curtate,scanning electron microscope analysis showed that pseudopodiao of A431 cells obviously decurtated and amount reduced,cell cycle progression analysis which performed by flow cytometry indicated that cell cycle was obviously inhibited by baicalein with it＇s dosage increasing and action time prolonging,and apoptosis ratio was also increased,it is significant to compare with control（P 0.05）.【Conclusion】 ①The optimal dosage of baicalein was 10μM to 30μM which inhibited A431 cell proliferation and without cytotoxicity after 48 hours treated by baicalein.②Baicalein suppressed A431 cells proliferation,cell cycle progression,pseudopod formation and tumor formation,so as to achieve anti-tumor hyperplasia and infiltrating-metastasis.