作者姓名：周小芸 王艳红 薛琼 孙瑞霞 陈洁 陈军 韩丹
    作者单位：复旦大学附属中山医院肝癌研究所
    中文关键字：肝癌；维生素K2；caspase-3；凋亡；转移
    英文关键字：Hepatocellular carcinoma; Vitamin K2; Caspase-3; Apoptosis; Metastasis

    中文摘要：目的：研究维生素K2对人肝癌细胞的凋亡诱导作用，并探讨其作用机制；研究维生素K2的干预对荷瘤裸鼠肝癌肺转移及生存期的影响。方法：用流式细胞仪检测细胞凋亡率；RT-PCR方法检测凋亡相关基因caspase-3、bcl-2及bax的mRNA表达。建立人肝癌裸鼠原位移植模型，给予荷瘤裸鼠口服维生素K2 (30 mg·kg-1·d-1)，另设对照组，观察两组裸鼠肝癌肺转移和生存期变化。结果：100 μmol·L-1的维生素K2与细胞共育6 h后细胞的凋亡率为（28.5±1.6）%，与对照组（2.8±4.8）%比较，有显著差异(P<0.05)。caspase-3 mRNA的表达随着维生素K2浓度及作用时间的增长而增强；当100 μmol的维生素K2分别作用细胞24 h和72 h后，caspase-3 mRNA的表达分别为(0.495±0.250)和(0.603±0.098)，与对照组比较(0.270±0.132)均有显著差异（P<0.05）。人肝癌细胞中未检测到bcl-2 mRNA表达。bax mRNA的表达于用药前后无变化。维生素K2干预荷瘤裸鼠后，其生存期（83.6±5.18 d）较对照组(58.2±9.47 d)明显延长(P<0.05)；其肺转移灶 (4.6±1.95个) 较对照组(12±6.6个)明显减少(P<0.05)。结论：维生素K2对人肝癌细胞有凋亡诱导作用，凋亡相关基因caspase-3参与了凋亡的调控；维生素K2能减少裸鼠肝癌肺转移率，能延长荷瘤裸鼠的生存期。

    英文摘要：Objective: To investigate the effects of vitamin K2 on apoptosis induction to HCC cells in vitro, and on pulmonary metastasis and life span to nude mice bearing human hepatocellular carcinoma in vivo.Methods: MHCC97H cells were cultured at the nutrient solution containing 100 μmol·L-1 of vitamin K2, following which the cell apoptosis rates were detected by flow cytometer at the 6th hours, and the expression of caspase-3, bcl-2 and bax mRNA were tested by reverse transcription-polymerase chain reaction (RT-PCR) assay. Nude mice bearing human hepatocellular carcinoma were treated with vitamin K2 (30 mg·kg-1·d-1), then their pulmonary metastases information was investigated with histological examination at the 6th weeks. Their life span was recorded.Results: When cells were treated with 100μmol·L-1 vitamin K2 for 6 h, the cell apoptosis rate was (28.5±1.6) % in vitamin K2 group and (2.8±4.8) % in control group, respectively. There was significant difference between the two groups, P<0.05. Increased expression of caspase-3 mRNA was in a dose- and time- dependent manner when the vitamin K2 dose increased from 20μmol·L-1 to 100μmol·L-1 and the time increased from 24h to 72h, respectively. The expression of caspase-3 mRNA showed significant difference between the vitamin K2 group (0.603±0.10) and the control group (0.270±0.13) following 72h of treatment, (P<0.01). Inversely, the expression of bcl-2 mRNA could not be detected, and bax mRNA did not show significant difference between the two groups.The pulmonary metastasis numbers was lesser in the vitamin K2 group (4.6±1.95) than that in the control group (12±6.60), P< 0.05. The life span was more prolonged in vitamin K2 group (83.6±5.18d) than that in control group (58.2±9.47d), P< 0.05. Conclusion: The apoptosis of MHCC97-H cells could be induced by vitamin K2 via caspase-3-transducted signal. Vitamin K2 also produced marked inhibitory effects on the pulmanory metastasis and beneficial influence on life span prolong of nude mice bearing human hepatocellular carcinoma.