汉防己甲素联合顺铂对NCI—H446细胞的影响及相关机制分析  

Effects and potential mechanisms of tetrandrine combined with cisplatin in NCI-H446 cell line

作　　者：许容容 李晓林[2] 叶伶云 胡松[1] 曾晓宁[1] 孔辉[1] 解卫平[1] 

机构地区：[1]南京医科大学第一附属医院呼吸科,210029 [2]南京医科大学老年消化科

出　　处：《国际呼吸杂志》2016年第4期247-253,共7页International Journal of Respiration

基　　金：国家自然科学基金（81273571）;江苏省呼吸病临床医学研究中心（BL2012012）

摘　　要：目的 研究汉防己甲素（tetrandrine，TET）联合顺铂（cisplatin，DDP）对人小细胞肺癌细胞株NCkH446体外抗肿瘤效果，探讨TET增强NCI-H446细胞对DDP敏感性的可能机制。方法体外培养人小细胞肺癌株NCI-H446；药物作用48h后，采用CCK-8法测定各组细胞活力；Edu染色观察各组细胞增殖情况；Hoechst染色检测各组细胞凋亡情况；Westernblot检测各组磷酸化Akt、Bax、Bcl-2、cleaved—caspase-3、LC3等蛋白表达水平变化。结果低浓度的TET与DDP联合后可有效抑制NCI—H446细胞株的增殖，诱导其凋亡；联合组半数抑制浓度较单药组显著降低；联合组磷酸化Akt表达显著下降，Bax、cleaved—caspase-3表达显著提高，抗凋亡蛋白Bcl-2表达下降；自噬相关蛋白LC3表达明显上调。结论低浓度TET与顺铂具有协同效应，其机制可能是通过上调促凋亡蛋白Bax，下调抗凋亡蛋白BcL2来诱导细胞凋亡；并且自噬可能在其中发挥重要作用。Objective To investigate the anti-tumor activity of tetrandrine （TET） combined with cisplatin （DDP） in human small cell lung cancer cell line NCI-H446, to study the potential mechanism of TET enhancing the sensitivities of DDP on NCI-H446. Methods Human small cell lung cancer cell line NCI-H446 was routinely cultured in vitro. Cell viability of each group was analyzed with CCK-8 assay after treated with different drugs for 48 hours. The proliferation rate was determined by Edu staining. Hoechst staining was used to evaluate the apoptosis cells of NCI-H446. The expressions of the related proteins such as P-Akt, Bax, Bcl-2, cleaved-easpase-3, LC3 were detected by Western blot. Results The low concentration of TET combined with DDP exerted a synergistic effect on inhibiting proliferation and inducing apoptosis of NCI-H446 cell line. The IC50 of combination group was significantly lower than that of other groups. The phosphorylation level of Akt was decreased, and the levels of Bax and cleaved- caspase-3 were increased, the expression of anti-apoptosis protein Bcl-2 was decreased in the combination group. The autophagy related protein LC3 was up-regulated simultaneously. Conclusions The low concentration of TET has synergistic effect with DDP via inducing apoptosis of NCI H446 by upregulating pro-apoptosis protein Bax and down-regulating anti apoptosis protein Bcl-2, and autophagy may play an important role in the apoptosis.

关 键 词：汉防己甲素 顺铂 协同效应 凋亡 自噬 

分 类 号：R734.2[医药卫生—肿瘤]