南赤瓟对Caski细胞的影响以及Survivin与Bcl-2蛋白家族基因表达关系研究

Studys of the inhibition effects from Thldiantha nudiflora Hemsl on Caski cells and survivin gene expression and Bcl-2 protein family relationships

目的:研究南赤瓟醇提物对Caski细胞抑制作用及可能的作用机制。方法:用不同浓度的南赤瓟醇提物(0.0625、0.125、0.25、0.50、1.00、2.00mg/ml)作用Caski细胞24h后,采用MTT法检测不同浓度的南赤瓟醇提物对Caski细胞增殖率的影响;用0.25、0.50和1.00mg/ml的南赤瓟醇提物和紫杉醇4×10-5mg/ml作用Caski细胞24h后,MTT法检测其对细胞活力的影响,流式细胞术和Hoechst 33258染色检测其对细胞凋亡的影响,实时定量PCR和Western blot检测Survivin、Bcl-2、Bax的mRNA和蛋白表达,并计算Bcl-2与Bax的比值。结果:南赤瓟醇提物对Caski细胞的生长抑制作用呈明显的剂量依赖性;南赤瓟醇提物(0.25、0.50和1.00mg/ml)和紫杉醇(4×10-5mg/ml)处理能显著抑制Caski细胞增殖,促进其凋亡;上调Bax mRNA和蛋白表达,下调Survivin、Bcl-2 mRNA和蛋白表达及Bcl-2与Bax的比值。结论:南赤瓟醇提物可显著抑制Caski细胞增殖,促进细胞凋亡,其作用机制可能与其上调Caski细胞Bax mRNA和蛋白表达,下调Survivin、Bcl-2 mRNA和蛋白表达及Bcl-2与Bax的比值有关。

Objective： To investigate the inhibition effects of alcohol extraction from Thldiantha nudiflora Hemsl on Caski cells,and based on it,to we explored its possible mechanism. Methods： Caski cells were treated with different concentration of alcohol extraction from Thldiantha nudiflora Hemsl（ 0. 0625,0. 125,0. 25,0. 50,1. 00,2. 00 mg / ml） for 24h; After Caski cells were treated with alcohol extraction from Thldiantha nudiflora Hemsl（ 0. 25,0. 50 and 1. 00 mg / ml） and paclitaxel（ 4 × 10^-5mg / ml） at 24 h,the cell viability was determined by MTT assay,the apoptosis and apoptotic morphology were detected by flow cytometry and hoechst 33258 analysis; the expressions of survivin,Bcl-2and Bax mRNA and protein were detected by RT-PCR and western blot,and Bcl-2 / Bax ratio was analyzed. Results： The inhibitory effect of alcohol extraction from Thldiantha nudiflora Hemsl on Caski cells displayed a significant dose response; alcohol extraction from Thldiantha nudiflora Hemsl（ 0. 25,0. 50 and 1. 00 mg / ml） and paclitaxel（ 4 × 10^-5mg / ml） might significantly promote apoptosis（ P〈0. 05,P〈0. 01,respectively）; upregulate the mRNA and protein expression levels of Bax,downregulate expression levels of survivin,Bcl-2 and Bcl-2 / Bax ratio compared with the negative control group（ P〈0. 05,P〈0. 01,respectively）. Conclusion： Alcohol extraction from Thldiantha nudiflora Hemsl could effectively inhibit proliferation and induce apoptosis in Caski cells. The mechanism may be related to upregulating the Bax mRNA and protein expressions,downregulating the survivin,Bcl-2 mRNA and protein expressions,decreasing Bcl-2 / Bax ratio.