论文部分内容阅读
目的观察芝麻素及联合吉西他滨对人肺腺癌A-549细胞株的作用。方法体外培养A-549细胞株。MTT法检测细胞增殖,Hoechst33258荧光染色法观察细胞形态学变化,流式细胞术检测细胞内Ca2+浓度变化,免疫细胞化学法观察caspase-3、caspase-8和caspase-9的表达。结果 20~80μg·mL-1芝麻素对A-549细胞均有一定的增殖抑制作用(P<0.01),而随着浓度增加,至70μg·mL-1时,增殖抑制作用逐渐减弱,IC50浓度为40μg·mL-1。0.001~20μg·mL-1吉西他滨对A-549细胞的生长均具有抑制作用(P<0.01),且呈现浓度依赖性,IC50浓度为1.0μg·mL-1;40μg·mL-1芝麻素+1.0μg·mL-1吉西他滨组的增殖抑制作用更为显著(P<0.01)。40μg·mL-1芝麻素组、1.0μg·mL-1吉西他滨组和芝麻素+吉西他滨组作用A-549细胞48 h后在荧光染色荧光显微镜下观察发现受抑制细胞核浓缩,细胞核染色质高度凝聚、边缘化,与对照组相比细胞内Ca2+浓度水平增高,caspase-3、caspase-8和caspase-9表达均显著增加(P<0.05),联用组均显著高于两药单用组(P<0.01)。结论芝麻素联合吉西他滨可协同抑制A-549细胞,其机制可能与提高细胞浆内Ca2+浓度进而诱导细胞凋亡,激活caspase通路有关。
Objective To observe the effect of sesamin and gemcitabine on human lung adenocarcinoma A-549 cell line. Methods A-549 cells were cultured in vitro. Cell proliferation was detected by MTT assay. Cell morphology was observed by Hoechst 33258 staining. The intracellular Ca2 + concentration was measured by flow cytometry. The expressions of caspase-3, caspase-8 and caspase-9 were detected by immunocytochemistry. Results Sesamin inhibited the proliferation of A-549 cells at a dose of 20 ~ 80μg · mL-1 (P <0.01), but inhibited the proliferation of the cells at 70μg · mL-1 with the increase of concentration. The inhibitory effect of gemcitabine on the growth of A-549 cells was inhibited by 40μg · mL-1.0.001 ~ 20μg · mL-1 Gemcitabine in a concentration-dependent manner with IC50 of 1.0μg · mL-1 and 40μg · mL- mL-1 sesamin + 1.0μg · mL-1 gemcitabine group was more significant (P <0.01). A-549 cells were treated with 40μg · mL-1 sesamin, 1.0μg · mL-1 gemcitabine and sesamin + gemcitabine for 48 hours. Fluorescence staining showed that the cells were inhibited by nuclear condensation, highly condensed nuclear chromatin, Compared with the control group, Ca2 + concentration increased and the expression of caspase-3, caspase-8 and caspase-9 significantly increased (P <0.05) in the combination group compared with the control group <0.01). Conclusion Sesamin and gemcitabine can inhibit A-549 cells synergistically. The mechanism may be related to increasing intracellular Ca2 + concentration and inducing apoptosis and activating caspase pathway.