热疗联合紫杉醇对人舌鳞癌细胞系CAL-27增殖、凋亡和周期影响

Effect of hyperthermia combined with paclitaxel on proliferation,apoptosis and cycle of human tongue squamous cell carcinoma cell line CAL-27

目的观察热疗联合紫杉醇对人舌鳞癌细胞系CAL-27增殖、凋亡和周期的影响并探讨其机制。方法CCK-8法确定紫杉醇工作浓度,将体外培养CAL-27细胞分为对照组、紫杉醇组、42℃热疗组和联合治疗组。克隆形成实验检测细胞增殖能力,流式细胞术检测细胞周期及凋亡,蛋白印迹法检测AKT、p-AKT、Bcl-2、Bax蛋白表达。结果紫杉醇组、热疗组及联合治疗组相比对照组均可抑制细胞增殖及促进凋亡(P<0.05),且联合治疗组优于热疗组、紫杉醇组(P<0.05)。蛋白印迹法实验结果显示联合治疗组上调Bax蛋白表达水平,同时下调P-AKT、Bcl-2表达(P<0.05)。结论42℃热疗与紫杉醇联合应用可抑制CAL-27细胞增殖及促进凋亡,其机制可能与抑制AKT活化及激活Bax/Bcl-2凋亡通路有关。

Objective To observe the effect of hyperthermia combined with paclitaxel on the proliferation,apoptosis and cycle of human tongue squamous cell carcinoma cell line CAL-27,and to explore the underlying mechanism.Methods The working concentration of paclitaxel was determined by CCK-8 assay,and the cultured CAL-27 cells were divided into the control,paclitaxel,42℃ hyperthermia and combined treatment groups.The ability of cell proliferation was detected by colony formation assay,and the cell cycle and apoptosis were determined by flow cytometry.The expression levels of AKT,p-AKT,Bcl-2 and Bax proteins in each group were measured by Western blot.Results Compared with the control group,the proliferation was significantly inhibited and the apoptosis of CAL-27 cells was significantly promoted in the combined treatment,hyperthermia and paclitaxel groups(all P<0.05),and the anti-proliferation and apoptosis-promoting effect in the combined treatment group was significantly better than those in the hyperthermia and paclitaxel groups(all P<0.05).Western blot showed that hyperthermia combined with paclitaxel could significantly up-regulate the expression level of Bax protein and significantly down-regulate the expression levels of P-AKT and Bcl-2 in CAL-27 cells(all P<0.05).Conclusions Hyperthermia combined with paclitaxel can play a synergistic role in inhibiting proliferation and promoting apoptosis of tongue squamous cell carcinoma CAL-27 cells.The mechanism may be related to the inhibition of AKT activation and the activation of Bax/Bcl-2 apoptosis signaling pathway.