三种黄芩提取物诱导人皮肤鳞状细胞癌SCL-1细胞凋亡的研究

Three Scutellaria balcalensls extracts induce the apoptosis of a human cutaneous squamous cell carcinoma cell line SCL-1： an experimental study

目的 探讨黄芩苷、黄芩素、汉黄芩素3种黄芩提取物对人皮肤鳞状细胞癌细胞株SCL-1细胞凋亡的影响.方法 将培养的SCL-1细胞分别用浓度为12.5、25、50、75、100μmol/L的3种黄芩提取物作用12、24、48 h后,噻唑蓝（MTT）法检测上述药物对SCL-1细胞的生长抑制作用.膜联蛋白V-异硫氰酸荧光素/碘化丙锭双染法和酶联免疫吸附试验（ELISA）检测SCL-1细胞凋亡率.流式细胞仪检测细胞周期.应用线性内差法,测定半数抑制浓度（IC50）;两样本均数比较用Student t检验;多样本均数比较采用方差分析,各样本之间多重比较采用SNK-q检验.结果 3种黄芩提取物均可抑制SCL-1细胞的生长,并具有浓度及时间依赖性（均P＜0.01）;在相同的药物浓度及作用时间下,黄芩素对SCL-1细胞生长抑制作用最强,黄芩苷抑制作用最弱,汉黄芩素抑制作用介于二者之间,差异具有统计学意义（均P＜ 0.01）.12.5 μmol/L黄芩苷、黄芩素、汉黄芩素分别作用SCL-1细胞48 h均可诱导SCL-1细胞凋亡并阻滞细胞周期的进程,其中G1期细胞比例分别为56.37％±177;2.41％、74.23％±177;2.02％、64.15％±177;1.87％,早期凋亡率分别为8.09％±177;1.02％、24.13％±177;0.76％、14.45％±177;1.57％,与对照组（45.04％±177;1.93％,4.12％±177;0.29％）比较,差异均有统计学意义（F=83.29,186.37,均P＜0.01）;其中黄芩素作用最强,其次为汉黄芩素,黄芩苷作用最弱.结论 黄芩能抑制SCL-1细胞生长并诱导其凋亡,为皮肤鳞状细胞癌的中药治疗开拓新的思路.

Objective To investigate the effect of three Scutellaria baicalensis extracts （baicalin,baicalein and wogonin） on the apoptosis of a human cutaneous squamous cell carcinoma cell line,SCL-1.Methods Methyl thiazolyl tetrazolium （MTT） assay was performed to detect the proliferation of cultured SCL-1 cells treated with different concentrations （12.5,25,50,75 and 100 μmol/L） of baicalin,baicalein and wogonin for various durations （12,24 and 48 hours）.Some SCL-1 cells were treated with baicalin,baicalein and wogonin of 12.5 μ mol/L respectively for 48 hours followed by the detection of cell apoptosis by double staining with annexin V-fluorescein isothiocyanate/propidium iodide in combination with enzyme-linked immunosorbent assay （ELISA）,as well as estimation of cell cycle by flow cytometry.The 50% inhibitory concentration was determined by line regression model and inner insert method,and statistical analysis was carried out by Student′s t test,one-way analysis of variance （ANOVA）,and Student-Newman-Keuls-q test.Results Baicalin,baicalein and wogonin all inhibited the proliferation of SCL-1 cells in a dose-and time-dependent manner （all P 〈 0.01）.Under the same conditions （treatment concentration and duration）,baicalein showed the strongest inhibitory effect on the proliferation of SCL-1 cells,followed by wogonin and baicalin （all P 〈 0.01）.All the Scutellaria baicalensis extracts induced the apoptosis of SCL-1 cells and arrested them in G1-phase.The percentage of cells in G1 phase was 56.37% ± 2.41%,74.23% ± 2.02% and 64.15% ± 1.87%,and early apoptosis rate was 8.09% ± 1.02%,24.13% ± 0.76% and 14.45% ± 1.57%,in SCL-1 cells treated with baicalin,baicalein and wogonin of 12.5 μmol/L for 48 hours,respectively,compared to 45.04% ± 1.93% and 4.12% ± 0.29% in the untreated control cells respectively （F =83.29,186.37,respectively,both P 〈 0.01）.Similarly,there was a downward trend from baicalein to wogonin and baicalin in the effect on cell apoptosis and cell cycle arrest of SCL-1 cells.Conclusions Scutellaria baicalensis can inhibit the growth and induce the apoptosis of SCL-1 cells,which may provide new ideas for the treatment of cutaneous squamous cell carcinoma with traditional Chinese drugs.