黄芩苷对错配修复基因缺失的结直肠癌裸鼠原位移植瘤细胞周期和凋亡的影响

Effects of baicalin on the apoptosis and cell cycle of colorectal cancer cells in orthotopic transplantation mice model with mismatch repair gene deficient

    摘要
  • 摘要:
    目的  研究黄芩苷对错配修复基因hMLH1缺失的结直肠癌HCT116细胞裸鼠原位移植瘤细胞周期和凋亡的影响。
    方法  建立人结直肠癌HCT116细胞裸鼠绿色荧光原位移植模型。根据随机数字表法将60只荷瘤裸鼠随机分为对照组和50、100、200 mg/kg黄芩苷组，每组15只。50、100、200 mg/kg黄芩苷组分别给予相应剂量的黄芩苷灌胃，2次/d；对照组采用等容量5%NaHCO3灌胃。应用流式细胞术检测肿瘤组织HCT116GFP细胞周期，TUNEL法观察细胞凋亡情况。组间比较采用方差分析或χ2检验，两两比较采用LSD-t检验。
    结果  裸鼠模型均构建成功，4组裸鼠体质量和肿瘤体积比较，差异无统计学意义(F=0.343，0.107，P>0.05)。50、100、200 mg/kg黄芩苷组结直肠癌细胞主要阻滞在G2/M期，其构成比分别为22%±6%、18%±7%和19%±6%，与对照组的7%±5%比较，差异有统计学意义(t=5.421，3.483，3.575，P<0.05)；50、100、200 mg/kg黄芩苷组间比较，差异无统计学意义(F=1.291，P>0.05)。50、100、200 mg/kg黄芩苷组细胞凋亡率分别为59.1%±10.0%、31.5%±9.3%、28.2%±10.8%，与对照组的2.3%±0.9%比较，差异有统计学意义(t=7.163，3.703，2.688，P<0.05)。50 mg/kg黄芩苷组结直肠癌细胞凋亡率显著高于200 mg/kg黄芩苷组(t=2.259，P<0.05)。
    结论  黄芩苷对错配修复基因hMLH1缺失的结直肠癌HCT116细胞裸鼠原位移植瘤生长具有明显的抑制作用。黄芩苷干预后移植瘤细胞周期在G2/M阻滞并诱导细胞凋亡，从而抑制HCT116细胞裸鼠原位移植瘤生长。

     

    Abstract:
    Objective  To study the effect of baicalin on the apoptosis and cell cycle of colorectal cancer cells in orthotopic transplantation mice model with mismatch repair gene hMLH1 deficient.
    Methods  Sixty orthotopic transplantation mice models of human colorectal cancer cell line HCT116 expressing green fluorescent protein (GFP) were established, and were randomly divided into the control group and the 50, 100, 200 mg/kg baicalin groups according to the random number table. Mice in the 50, 100, 200 mg/kg baicalin groups received intragastric infusion of baicalin at the corresponding dosages twice a day, while mice in the control group received intragastric infusion of 5% NaHCO3. Cell cycles and apoptotic rates of the HCT116GFP cells were detected by flow cytometry and TUNEL method respectively. Differences between the 2 groups were analyzed using the analysis of variance or chi-square test, and differences within each group were analyzed using the LSD-t test.
    Results  The orthotopic transplantation mice models of human colorectal cancer were successfully constructed, and there was no significant difference in the body weight of the mice and tumor size among the 4 groups (F=0.343, 0.107, P>0.05). The proportion of HCT116 GFP cells in the G2/M phase in the 50, 100, 200 mg/kg baicalin groups were 22%±6%, 18%±7% and 19%±6%，which were significantly higher than 7%±5% of the control group (t=5.421, 3.483, 3.575, P<0.05). There were no significant differences in the proportion of HCT116 GFP cells in the G2/M phase among the 50, 100, 200 mg/kg baicalin groups (F=1.291, P>0.05). The apoptotic rates of HCT116 GFP cells in the 50, 100, 200 mg/kg baicalin groups were significantly higher than the control group (t=7.163, 3.703, 2.688, P<0.05). The apoptotic rate of the 50 mg/kg baicalin group was significantly higher than that of the 200 mg/kg baicalin group (t=2.259, P<0.05).
    Conclusions  Baicalin significantly inhibits tumor growth in the orthotopic transplantation mice model with mismatch repair gene hMLH1 deficient. After treated with baicalin, the cell cycle is arrested at the G2/M phase, thus the tumor growth is inhibited.

     

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