金龙胶囊对结肠癌血管生成拟态及Mig-7的影响

刘娇萍; 袁昌劲; 余涛; 刘礼

doi:10.3971/j.issn.1000-8578.2016.12.010

金龙胶囊对结肠癌血管生成拟态及Mig-7的影响

Effect of Jinlong Capsule on Vasculogenic Mimicry and Mig-7 of Colon Cancer

摘要

摘要:
目的探讨金龙胶囊对结肠癌血管生成拟态（vasculogenic mimicry, VM）及迁移诱导蛋白7（Mig-7）的影响。
方法采用细胞三维培养技术观察HCT116、HT29结肠癌细胞株VM的形成能力。对VM（+）组细胞进行后续实验，实验分为空白组、中药组（金龙胶囊）和阳性对照组（200 μmol/L, 5-氟尿嘧啶组），其中金龙胶囊组按培养液浓度分为低浓度（0.01 mg/ml）、中浓度（0.02 mg/ml）和高浓度组（0.04 mg/ml）三组。采用RT-PCR和Western blot法分别检测各组Mig-7 mRNA和Mig-7蛋白的表达。
结果 HCT116细胞可形成VM，HT29细胞无形成VM能力；Mig-7在HCT116细胞中阳性表达；金龙胶囊高、中、低剂量及5-氟尿嘧啶组均可干扰HCT116细胞VM形成，下调Mig-7 mRNA和蛋白的表达，各药物组VM及Mig-7的表达量均低于空白对照组，差异有统计学意义（P<0.05），随着金龙胶囊剂量增大，VM及Mig-7的表达量逐渐减少。以金龙胶囊高剂量组（0.04 mg/ml）和5-氟尿嘧啶组效果更为明显（P<0.01），且5-氟尿嘧啶组效果优于金龙胶囊高剂量组（0.04 mg/ml）（P<0.05）。
结论 VM（+）组细胞可表达Mig-7 mRNA和Mig-7蛋白，金龙胶囊抑制VM的形成，可能与下调Mig-7 mRNA和Mig-7蛋白的表达有关。

Abstract:
Objective To explore the effect of Jinlong capsule on vasculogenic mimicry and Mig-7 of colon cancer.
Methods To observe the capacity of forming VM about HCT116 colon cancer cells and HT29 colon cancer cells when cultured in three dimensional condition. The VM (+) cells for subsequent experiments, which were divided into blank group, Chinese medicine group (Jinlong capsule) and positive control group (5-fluorouracil, 200μmol/L). The Jinlong capsule culture was divided into low dose group (0.01mg/ml), medium dose group (0.02mg/ml) and high dose group (0.04mg/ml). The expression of Mig-7 mRNA and protein were tested by RT-PCR and Western blot, respectively.
Results HCT116 colon cancer cells could form visible VM structure and express Mig-7. The high, medium, and low dose Jinlong Capsule and 5-fluorouracil group could inhibit the capacity of forming VM about HCT116 colon cancer cells and reduce the expression of Mig-7 mRNA and protein. The expression of VM and Mig-7 of each treated group were lower than that in the blank control group, the difference was statistically significant(P<0.05). The VM and Mig-7 expression gradually reduced with the increase of Jinlong capsule dose. The effect of Jinlong capsule high dose group (0.04 mg/ml) and 5-fluorouracil group were superior to others(P<0.01), and 5-fluorouracil group was better than Jinlong capsule high-dose group (0.04mg/ml) (P<0.05).
Conclusion The VM(+) colon cancer cells could express Mig-7 mRNA and protein. Jinlong capsule can inhibit the formation of VM probably by reducing the expression of Mig-7 mRNA and protein.

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