摘要
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。M1Tr法检测细胞增殖．Hoeehst33258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1．0Ixg·mL-1；40μg·mL。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48h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Can浓度水平增高，caspase-3、caspase-8和caspase．9表达均显著增加（P〈O．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Ca卦浓度进而诱导细胞凋亡，激活caspase通路有关。
AIM To observe the effect of sesamin with gemcitabine on human lung adenocarcinoma cell line A-549. METHODS Human lung adenocarcinoma cell line A-549 was cultured in vitro. Cell proliferation was assayed by MTT, and morphological changes of cell were observed by Hoechst33258 flourescence staining, and the change of the concentration of intracellular Ca2＋ was detected by flow cytometry. The expression of caspase-3, caspase-8 and caspase-9 was observed by immunocytochemistry. RESULTS Sesamin 20 - 80 μg·mL-1 had certain depressant effect on proliferation of A-549 （P 〈 0.01）. But when the concentration increased to 70μg·mL-1, the depressant effects of sesamin on cell proliferation gradually weakened. IC50 of sesamin was 40 μg·mL-1 Gemcitabine 0.001 - 20 μg·mL-1 had inhibitory action on growth of A-549 （P 〈 0.01）, and showed concentration dependent. IC50 of gemcitabine was 1.0 μg·mL-1. The depressant effect on cell proliferation of sesamin 40μg·mL-1 combined with gemcitabine 1.0μg·mL-1 was more significant （P 〈 0.01）. After 48 h treatment, sesamin 40 μg·mL-1 group, gemcitabine 1.0μg·mL-1group and sesamin combined gemcitabine group were observed inhibited nucleolus condensed, nuclear chromatin condensed and marginalized by fluorescence microscope. Compared with the control group, Ca2＋ level of intra-cellular, expression of caspase-3, caspase-8 and caspase-9 was significantly increased （P 〈 0.05）, and the combination group was significantly higher than either of the single-use drug group （P 〈 0.01）. CONCLUSION Sesamin combined with gemcitabine can synergistically inhibit A- 549, and the mechanism may be induce apoptosis by increasing the Ca2＋ concentration of intracytoplasm, and activation of caspase pathway.
出处
《中国新药与临床杂志》
CAS
CSCD
北大核心
2014年第7期529-533,共5页
Chinese Journal of New Drugs and Clinical Remedies
