【摘要】 目的:探讨马尾松树皮提取物(Pinus massoniara bark extract,PMBE)在体外抑制肺腺癌GLC-82细胞生长的机制。方法:通过CCK-8法分析了PMBE对GLC-82细胞的生长活力影响;AO/EB染色、透射电镜和流式细胞术检测了PMBE诱导的肺腺癌细胞GLC-82的凋亡;Western blot检测PMBE处理GLC-82细胞后caspase-8、caspase-9、caspase-3、Bcl-2和Bid蛋白表达水平的变化。结果:PMBE在体外抑制肺腺癌GLC-82细胞的生长具有时间和剂量依赖性;通过荧光显微镜、透射电镜和流式细胞术证实PMBE诱导了GLC-82细胞的凋亡;细胞周期分析结果说明PMBE可以显著减少S期的GLC-82细胞;免疫杂交发现PMBE可以诱导caspase-3、caspase-8、caspase-9蛋白的激活,并下调Bcl-2和Bid蛋白的表达。结论:PMBE可能通过激活caspase-8从而激活Bid蛋白,进一步激活caspase-9来诱导肺腺癌GLC-82细胞的凋亡。更多还原

【Abstract】 Objectives: To research the inhibitory effects of Pinus massoniana bark extract(PMBE) on the lung adenocarcinoma GLC-82 cell in vitro. Methods: The CCK-8 method was used to analyze the inhibitory effect of PMBE on the growth of GLC-82 cells. The apoptosis of GLC-82 cell was measured by AO/EB staining, transmission electron microscopy, and flow cytometry. The expressions of caspase-8, caspase-9, caspase-3, Bcl-2 and Bid proteins were detected by Western blot method. Results: The growth of GLC-82 cells could be inhibited by PMBE that dependent on the concentration and time. PMBE could induce the apoptosis of GLC-82 cells that was confirmed via fluoroscope, TEM and flow cytometry. Cell cycle analysis revealed that the number of GLC-82 cells in the S phase was decreased by the treatment of PMBE. The result of Western blot showed that both the extrinsic and intrinsic pathways of the caspase cascade were activated, including caspase-8, caspase-9 and caspase-3. In addition, the expression of the Bcl-2 and Bid proteins were decreased. Conclusion: PMBE could induce the apoptosis of GLC-82 cells via activating caspase-8 to cleave the intact Bid that was an enzyme to activate caspase-9.更多还原