健脾解毒方对人结肠癌多药耐药细胞的逆转作用

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.

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

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.