健脾解毒方对人结肠癌多药耐药细胞的逆转作用

Reversal Effect of Jianpi Jiedu Formula on Multidrug Resistance in Human Colon Carcinoma Cells

目的:探讨健脾解毒方对人结肠癌多药耐药细胞HCT8/V的逆转作用及其可能机制。方法:人结肠癌HCT8/V多药耐药细胞常规培养,5%健脾解毒方药物血清作用48 h后,MTT法检测HCT8/V细胞对化疗药敏感性的影响,高效液相色谱法(HPLC)检测耐药细胞内长春新碱(VCR)药物浓度,流式细胞仪分析健脾解毒方对5-氟尿嘧啶(5-Fu)诱导的HCT8/V细胞凋亡和细胞周期的影响。结果:健脾解毒方能增加HCT8/V细胞对VCR、顺铂(DDP)、5-Fu、吡柔比星(THP)4种化疗药物的敏感性,4种化疗药的IC50分别从(191.08±18.18)mg·L-1下降到(90.13±6.33)mg·L-1;(290.79±28.38)mg·L-1下降到(22.16±1.82)mg·L-1;(23.12±1.99)mg·L-1下降到(9.88±0.86)mg·L-1;(26.40±2.92)mg·L-1下降到(19.41±1.48)mg·L-1;健脾解毒方药物血清作用HCT8/V细胞48 h后细胞内VCR浓度明显增高(P<0.01),且呈剂量依赖性。健脾解毒方能提高5-Fu诱导的HCT8/V细胞的凋亡率,使细胞阻滞于G1期。结论:健脾解毒方通过增加结肠癌多药耐药细胞内化疗药物浓度,抑制细胞增殖,诱导其凋亡,逆转HCT8/V细胞株的多药耐药性。

Objective： To investigate the mechanisms and reversal effects of Jianpi Jiedu formula on human colon carcinoma multidrug resistance（MDR） in HCT8/V cells.Method： Human colon carcinoma MDR cells were cultured conventionally.The concentration of Jianpi Jiedu formula（5%） combined with chemotherapy drugs and the reversal effect of multidrug resistance（MDR） in HCT8/V cells was evaluated by the MTT assay.The variation of intracellular concentration of vincristine（VCR） in HCT8/V cells was evaluated by the HPLC,and the effects of Jianpi Jiedu formula on 5-Fu induced apoptosis and cell cycle were evaluated by flow cytometry（FCM）.Result： Jianpi Jiedu formula could increase the sensitivity of HCT8/V cells to VCR,DDP,5-Fu,THP.The IC50 value of VCR decreased from（191.08±18.18） mg·L-1 to（90.13±6.33） mg·L-1,DDP was from（290.79±28.38） mg·L-1 to（22.16±1.82） mg·L-1,5-Fu was from（23.12±1.99） mg·L-1 to（9.88±0.86） mg·L-1,and THP was from（26.40±2.92） mg·L-1 to（19.41±1.48） mg·L-1.HPLC results showed that Jianpi Jiedu formula increased the concentration of drugs in HCT8/V cells in a dose-dependent manner after treatment for 48 h.Flow cytometry assay result verified that the inhibited effects of Jianpi Jiedu formula combined with 5-Fu on the proliferation of HCT8/V cells was closely related to the arrest in G1 phase and increasing of apoptosis.Conclusion： Jianpi Jiedu formula can reverse MDR in HCT8/V cell,which possibly through inhibiting cell proliferation,inducing cell-apoptosis,ultimately affect the concentration of drugs inside the cells.