氯化两面针碱体外对人口腔鳞癌多药耐药细胞KBV200的抗癌活性

Anticancer activity of nitidine chloride from Zanthoxylum nitidum(Roxb.)DC.on multidrug resistant KBV200 cells in vitro

目的探讨氯化两面针碱对多药耐药癌细胞的抗癌作用。方法应用MTT比色法测定细胞存活率,流式细胞术测定细胞周期,Hoechst 33258染色法检测细胞凋亡,酶联免疫吸附法测定半胱氨酸天冬氨酸蛋白酶3(caspase 3)含量。结果氯化两面针碱(0.75,1.5,3,6和12mg·L-1)浓度依赖性地降低人口腔鳞癌多药耐药细胞KBV200的存活率,作用48h时IC50值为(2.43±0.19)mg·L-1;氯化两面针碱(0,3,6和9mg·L-1)与KBV200细胞作用48h,细胞凋亡率依次为(1.6±0.4)%,(2.3±0.7)%,(14.0±2.5)%和(8.3±2.2)%;氯化两面针碱6mg.L-1可使KBV200细胞caspase3含量升高,3mg·L-1作用12,24和48h可增加KBV200细胞G2/M期细胞比例。结论氯化两面针碱对多药耐药KBV200细胞具有抑制生长、促进凋亡和阻滞细胞周期的作用,可能是对多药耐药癌细胞敏感的一种中药活性成分。

AIM To explore the anticancer activity of nitidine chloride from Zanthoxylum nitidum （Roxb.） DC. on muhidrug resistant KBV200 ceils. METHODS MTT assay was used to examine the survival rate of ceils. Flow cytometry was performed to measure the cell cycle. Hoechst 33258 staining was adopted to detect cell apoptosis. Enzyme-linked immunosor-bent assay was employed to detect the content of caspase 3. RESULTS Nitidine chloride （0.75, 1.5, 3, 6 and 12 mg·L^-1） decreased the survival rate of KBV200 ceils in a concentration-dependent manner, the IC50 value for 48 h was （2.43± 0.19 ） mg· L^- 1 When KBV200 cells were incubated with nitidine chloride 0, 3, 6 and 9 mg·L^-1, respectively, for 48 h the ratios of apoptotic ceils were （ 1.6 ±0. 4 ） %, （2.3 ±0.7）% ,（14.0 ±2.5）% and （8.3±2.2 ） %, respectively. When nitidine chloride was 6 mg·L^-1, the content of caspase 3 in KBV200 cells was higher compared with control. Nitidine chloride 3 mg·L^-1 increased the percentages of G2/M cells when incubated with KBV200 cells for 12, 24 and 48 h, respectively. CONCLUSION Nitidine chloride inhibits the growth, promotes apoptosis and arrests the process of cell cycle of KBV200 ceils. It might be an active component sensitive to multidrug resistant tumor ceils.