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目的:探讨维生素C和维生素K3联合对诱导人肝癌细胞的Autoschizis现象。方法:采用体外培养技术,MTI法观察细胞生长抑制作用;普通光学显微镜、透射电镜及激光扫描共焦显微镜下分别观察细胞形态,流式细胞仪检测细胞凋亡和细胞周期分布。结果:当维生素C(100、250、400、500μmnol/L),维生素K3(2、5、8、10μgmol/L)单独作用时,对肝癌细胞的生长影响并不明显(P>0.05),而维生素C和维生素K3以生理剂量联合后,能显著抑制肝癌细胞Hep G2的生长。维生素C和维生素K3作用于细胞后,可看到较为典型细胞凋亡的形态学变化:细胞核固缩,染色质凝集,呈新月型紧贴于核膜周边,核碎裂,染色质片断化,空泡形成和凋亡小体等。流式细胞仪DNA直方图上出现典型的亚二倍体“凋亡峰”,表现为细胞周期特异性,作用72 h后,与对照组比较,G0/G1期细胞上升,S期细胞下降,细胞周期阻滞于G0/G1期。结论:维生素C和维生素K3抑制肝癌细胞增殖和诱导细胞凋亡的作用明显,而从形态中未观察到Autoschizis特殊形式。维生素C和维生素K3可通过诱导肝癌细胞凋亡,而具有治疗肝癌的潜在价值。
Objective: To investigate the Autoschizis phenomenon induced by the combination of vitamin C and vitamin K3 on human hepatoma cells. Methods: The cell growth inhibition was observed by MTI method in vitro. The cell morphology was observed under ordinary light microscope, transmission electron microscope and laser scanning confocal microscope. The apoptosis and cell cycle distribution were detected by flow cytometry. Results: The effects of vitamin C (100, 250, 400, 500μmnol / L) and vitamin K3 (2,5,8,10μgmol / L) alone were not significant (P> 0.05) Vitamin C and vitamin K3 combined with physiological doses, can significantly inhibit the growth of Hep G2 cells. Vitamin C and vitamin K3 role in the cells, we can see the more typical morphological changes of apoptosis: nuclear condensation, chromatin condensation, crescent-type close to the nuclear perimeter, nuclear fragmentation, chromatin fragmentation , Vacuolar formation and apoptotic bodies. Flow cytometry DNA histogram showed a typical sub-diploid “apoptosis peak”, showing the cell cycle-specific, after 72 h, compared with the control group, G0 / G1 phase cells increased, S phase cells decreased, Cell cycle arrest in G0 / G1 phase. Conclusion: The effects of vitamin C and vitamin K3 on the proliferation and apoptosis of hepatocellular carcinoma cells were obvious, but no special form of Autoschizis was observed from the morphology. Vitamin C and vitamin K3 have the potential value of treating liver cancer by inducing hepatoma cell apoptosis.