虫草素增强顺铂诱导的食管癌细胞系Eca109凋亡

Cordycepin enhances apoptosis of Eca109 cells induced by cis-platinum

目的研究虫草素与顺铂联合用药对食管癌细胞Eca109凋亡的影响及其作用机制。方法将食管癌细胞Eca109分为对照组、不同浓度虫草素处理组、不同浓度顺铂处理组和虫草素(70μg/m L)与顺铂(0.8μg/m L)联合处理组。MTS法检测Eca109细胞增殖;Hoechst 33258染色法及流式细胞术检测Eca109细胞凋亡;Western blot法检测细胞核内NF-κB P65及凋亡相关蛋白Bcl-2和Bax表达水平;ELISA法检测细胞核内NF-κB P65与DNA的结合活性。结果虫草素与顺铂联合应用使顺铂对Eca109细胞的抑制率由29.30%增加至70.41%(P<0.05),增强了顺铂对Eca109的敏感性;与顺铂单独用药相比,联合用药能够显著增加顺铂诱导的细胞凋亡(P<0.05);与对照组相比顺铂能够增加细胞核内NF-κB P65的活性,而虫草素却能抑制NF-κB P65的活性,联合用药后NF-κB P65的活性下调;与虫草素和顺铂单独用药相比,联合用药组能够使抗凋亡蛋白Bcl-2的表达显著降低(P<0.05),而促凋亡蛋白Bax表达则显著增高(P<0.05)。结论虫草素可能通过抑制NF-κB途径,调节下游信号分子Bcl-2和Bax的表达,增强顺铂对Eca109的凋亡诱导效应。

Objective To investigate the mechanism of action and impact on apoptosis of Eca109 treated with cordycepin plus cisplatin. Methods Divide Eca109 into four groups as control group,cordycepin,cisplatin and combined group which was treated with different concentrations of cordycepin,cisplatin and cordycepin（ 70 μg / m L）and cisplatin（ 0. 8 μg / m L）. The proliferation was measured by MTS assay. Apoptosis was observed by using Hoechst 33258 staining and flow cytometry. Western blot was used to detect the expression of NF-κB P65,Bcl-2 and Bax. Enzyme linked immunosorbent assay（ ELISA） was used to detct the binding activity with NF-κB P65 and DNA in cell nucleus. Results The inhibition ratio induced by cordycepin and cis-platinum combination was increased from 29. 3% to 70. 4% in Eca109 cells（ P〈0. 05）. Cordycepin combined with cis-platinum can remarkablely promote apoptosis of Eca109 cells. Cisplatin increased the activity of the NF-κB P65 in the nuclei while cordycepin can inhibit the activity of the NF-κB P65,the activity of the NF-κB P65 decreased with combined treatment of cordycep and cisplatin. Compared to cisplatin alone,cordycepin combined with cisplatin decreased the expression of Bcl-2 while remarkblely increased the expression of Bax. Conclusions Cordycepin could enhance the apoptosis of Eca109 cells induced by cisplatin and regulated downstream signaling molecule expression of Bcl-2and Bax by inhibiting the NF-κB pathway.