摘要
目的 研究三七总皂苷（PNS）对K562细胞增殖、凋亡、周期的影响及其相关分子机制。方法采用MTT法检测PNS对K562细胞增殖的影响；AO／EB双荧光染色法及Annexinv-FITc／PI双染色法观察细胞凋亡及死亡状况；流式细胞术检测K562细胞的周期变化；RT-PCR检测mTOR信号通路主要分子mRNA的表达变化；Westernblot检测cleavedcaspase-3蛋白、cyclinD1蛋白及mTOR信号通路主要蛋白及磷酸化蛋白的表达量变化。结果100—800rtg／mL的PNS作用于K562细胞后能够抑制细胞增殖。PNS能够上调cleavedcaspase-3蛋白表达，促使K562细胞发生凋亡。并且下调cyclinD1蛋白表达，促使K562细胞阻滞在G0/G1期。同时，PNS能够抑制K562细胞mTORmRNA表达，降低mTOR蛋白和磷酸化蛋白p-mTOR（Ser2448）、p-p70S6K（Thr229／389）及P-4E-BP1（Thr37／46）的表达，从而抑制mTOR信号通路活性。结论PNS对体外培养K562细胞有抑制增殖，促进凋亡，使其发生周期阻滞的作用。其机制可能与PNS抑制mTOR信号通路活性、上调cleavedcaspase．3蛋白表达及抑制cyclinD1蛋白表达有关。
Objective To investigate the effects of Panax notoginseng saponins （PNS） on the proliferation, apoptosis and cell cycle of K562 cells and explore the molecular mechanisms underlying these effects. Methods PNS-induced growth inhibition of K562 cells was detected by MTT assay; the cell apoptosis was evaluated by AO/EB staining and Annexin V-FITC/PI staining; flow cytometry was used to detect cell cycle changes in the treated cells. The mRNA expressions of the molecules in roTOR signaling pathway were examined by RT-PCR, and the cellular expressions of cleaved caspeas-3, cyclin D1 and major proteins in mTOR signaling pathway were detected using Western blotting. Results MTT assay showed that treatment with 100-800 μg/mL PNS significantly inhibited the proliferation, promoted the cell apoptosis, and caused cell cycle arrest in G0/G1 phase in K562 cells. Western blotting revealed increased protein expression of cleaved caspase-3 and decreased expression of cydin D1 in PNS-treated ceils, in which the proteins expressions of mTOR, p-roTOR, p-p70S6K and p4E-BP1 and the mRNA expression of mTOR were all decreased： Conclusion PNS can inhibit the proliferation, induce apoptosis and cause cell cycle arrest in K562 cells possibly by up-regulating cleaved caspase 3 and down-regulating cyclin D1 and mTOR signaling pathway.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2015年第8期1103-1109,共7页
Journal of Southern Medical University
基金
国家自然科学基金(81000992)~~
关键词
三七总皂苷
K562细胞
增殖
凋亡
周期
K562
Panax notoginseng saponins
proliferation
apoptosis
cell cycle
