法卡林二醇单药及与其他药物联用对乳腺癌细胞凋亡的影响

Effects of falcarindiol alone and combination with other drugs on apoptosis of breast cancer cells

目的探讨法卡林二醇(FAD)单药及与其他药物联用对乳腺癌细胞凋亡的影响。方法选择乳腺癌细胞MDA-MB-231、MDA-MB-468及正常乳腺细胞MCF-10A,每种细胞分为5组,分别加入0、3、6、12、24μmol/L FAD作用24 h,采用MTT法检测细胞增殖率。结果显示,两种乳腺癌细胞在0、3、6、12μmol/L FAD作用下细胞增殖率逐渐降低(P均<0.05);24μmol/L FAD作用下细胞增殖率低于0、3、6μmol/L FAD(P均<0.05),但与12μmol/L FAD作用下比较P>0.05。考虑到干预效果及节省试剂的原则,我们选择3μmol/L FAD进行后续研究。取对数生长期三种细胞,每种细胞随机分为FAD组和对照组,FAD组加入含3μmol/L FAD的培养基培养,对照组常规培养,培养24 h,收集细胞,采用流式细胞仪检测细胞凋亡率。选择乳腺癌细胞MDA-MB-231,随机分为空白组、5-氟尿嘧啶组(5-Fu组)、FAD组、5-Fu+FAD组。空白组常规培养,5-Fu组加入含25μmol/L 5-Fu的培养基培养,FAD组加入含1μmol/L FAD的培养基培养,5-Fu+FAD组加入含25μmol/L 5-Fu及1μmol/L FAD的培养基培养,培养24 h,收集细胞,采用流式细胞仪检测细胞凋亡率。将25μmol/L 5-Fu换为10 nmol/L硼替佐米(BRZ),重复上述试验。结果两种乳腺癌细胞的FAD组细胞凋亡率均高于其对照组(P均<0.01),MCF-10A细胞的FAD组细胞凋亡率与其对照组比较P>0.05。5-Fu组、FAD组、5-Fu+FAD或BRZ+FAD组细胞凋亡率均高于空白组(P均<0.01),5-Fu+FAD组或BRZ+FAD组细胞调亡率均高于5-Fu组和FAD组(P均<0.01),而5-Fu组与FAD组比较P>0.05。结论 FAD可促进乳腺癌细胞凋亡并抑制其增殖,但不影响正常乳腺细胞;FAD联合5-Fu或BRZ对促进乳腺癌细胞凋亡的效果优于FAD单药。

Objective To investigate the effects of falcarindiol（ FAD） alone and combination with other drugs on the apoptosis of breast cancer cells. Methods We selected the breast cancer cells MDA-MB-231,MDA-MB-468,and normal mammary cells MCF-10 A. The cells of each kind were divided into five groups,which were added with 0,3,6,12,and 24 μmol/L,respectively. The cell proliferation rate was detected by MTT. As the result,the proliferation rates of two breast cancer cells decreased gradually at 0,3,6,and 12 μmol/L FAD（ P〈0. 05）. The proliferation rate of cells under the treatment of 24 μmol/L FAD was lower than that under the treatment of 0,3,and 6 μmol/L FAD（ P〈0. 05）,and was also lower than that under 12 μmol/L FAD（ P〈0. 05）. Considering the effect of intervention and the principle of saving reagent,we selected 3μmol/L FAD for the further study. These three kinds of cells in the logarithmic growth were taken,and each kind of cells was divided into the FAD group and control group. The FAD group was cultured with 3 μmol/L FAD,and the control group was cultured as normal. All the cells were cultured for 24 h and the apoptosis rate was detected by flow cytometry. Breast cancer cells MDA-MB-231 were randomly divided into four groups： the blank group,5-fluorouracil group（ 5-Fu group）,FAD group,and 5-Fu ＋ FAD group. The blank group was normally cultured,the 5-Fu group was added with 25 μmol/L 5-Fu,the FAD group was added with 1 μmol/L FAD,and the 5-Fu ＋ FAD group was added with 25 μmol/L 5-Fu and 1 μmol/L FAD. All cells were cultured for 24 h and then were collected to detect the apoptosis rate by flow cytometry. The 25 μmol/L 5-Fu was replaced with 10 nmol/L bortezomib（ BRZ）. The test above was repeated. Results The apoptosis rate of two kinds of breast cancer cells in the FAD group was higher than that of the control group（ P〈0. 01）. There was no significant difference in the apoptosis rate between the FAD group and the control group（ P〈0. 05）. The apoptosis rates of the 5-Fu group,FAD group,5-Fu ＋ FAD group or BRZ ＋ FAD group were higher than that of the control group（ all P〈0. 01）. The apoptosis rate of 5-Fu ＋ FAD group or BRZ ＋ FAD was higher than that of the 5-Fu group and FAD group（ P〈0. 01）,but no significant difference was found between the 5-Fu group and FAD group（ P〉0. 05）. Conclusion FAD can promote the apoptosis of breast cancer cells and inhibit the cell proliferation but it dose not affect normal mammary cells. The effect of FAD combined with 5-Fu or BRZ on the apoptosis of breast cancer cells is better than that of FAD alone.