维生素C和维生素K3联合诱导肝癌细胞Autoschizis的探讨

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,MTT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:当维生索C(100、250、400、500 μmol/L),维生素K3(2、5、8、10μmol/L)单独作用时,对肝癌细胞的生长影响并不明显(P＞0.05),而维生采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,C0/G1期细胞上升,S期细胞下降,细胞周期阻滞于C0/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.

Inducting Activity of Vtamin C and Vitamin K3 on Hepatocarcinoma Cell Autoschizis

Objective:To explore the effect of vitamin C and vitamin K3 on induction of autoschizis hepatocarcinoma cell Hep G2. Methods: By cells culturing in vitro,anti-proliferation effects of vitamin C and vitamin K3 were assessed with methyl thiazolyl tetraznlium (MTT) colorimetric assay.The morphology of cells was observed under light microscope, transmission electron mircroscope and laser scanning confocal microscope respectively. The flow cytom-etry was used to detect apoptotic rate and cell cycle distribution. Results: Vitamin C and vitamin K3 separately did not induce any change of cell proliferation on Hep C2,but showed a remarkable anti-proliferation as both combined. Morphological changes characteristic of apoptosis were observed though light microscope, electron microscope:cellshrinkage,nuclear condensation,DNA fragmentation,vacuolation and formation bodies. A typical subdiploid peak before G0/G1 phase was observed by flow cytometry. The effect of vitamin C and vitamin K3 on Hep G2 showed a remarkable cell cycle spectficty, blocking at G0/ G1 phase. Conclusion: Vitamin C and vitamin K3 inhibit significantly proliferation of hepatocarcinoma cell Hep G2 and induce apoptosis, but no autoschizis. vitamin C and vitamin K3 may have a promising prespect in the treatment of hepatocarcinoma cancer though apoptosis.