甘露糖对乳癌细胞增殖及表柔比星化疗敏感度影响

EFFECT OF MANNOSE ON THE PROLIFERATION OF BREAST CANCER CELLS AND THEIR CHEMOSENSITIVITY TO EPIRUBICIN

目的探讨甘露糖对乳癌细胞增殖及表柔比星(EPI)化疗敏感性的影响。方法免疫印迹法测定乳癌细胞(BT-549、HCC1937、T-47D、MCF-7)和正常乳腺细胞(MCF-10A细胞)代谢甘露糖的关键蛋白磷酸甘露糖异构酶(MPI)的表达。将MCF-7和T-47D细胞随机分为对照组(含体积分数0.10胎牛血清的普通培养液)、甘露糖组(25 mmol/L甘露糖)、葡萄糖组(25 mmol/L葡萄糖)、EPI组(1μmol/L EPI)、甘露糖联合EPI组(1μmol/L EPI+25 mmol/L甘露糖)和葡萄糖联合EPI组(1μmol/L EPI+25 mmol/L葡萄糖),各组分别加入含相应药物培养液,于有氧条件和乏氧条件下培养,采用噻唑蓝(MTT)比色法测定甘露糖及甘露糖联合EPI对细胞生长活性的影响,免疫印迹法检测对照组、甘露糖及葡萄糖组T-47D细胞凋亡相关蛋白Caspase-3、Caspase-8、Bax和Bcl-2的表达。结果 MCF-10A、BT-549、HCC1937、T-47D和MCF-7细胞中MPI表达差异有统计学意义(F=392.680,P<0.05)。与MCF-10A细胞相比,T-47D细胞MPI表达量相对较低,MCF-7细胞表达相对较高,差异有统计学意义(t=25.449、32.857,P<0.05)。有氧条件下,甘露糖组T-47D细胞增殖率较对照组明显降低(t=33.338,P<0.05),MCF-7细胞增殖率与对照组比较差异无显著性(P>0.05);甘露糖联合EPI组T-47D细胞增殖率显著低于EPI单药组,差异有统计学意义(t=22.901,P<0.05);甘露糖联合EPI组与对照组相比MCF-7细胞增殖率差异无统计学意义(P>0.05)。乏氧条件下,甘露糖组T-47D细胞和MCF-7细胞增殖率较对照组显著降低(t=24.538、35.158,P<0.05),甘露糖联合EPI组T-47D和MCF-7细胞增殖率低于EPI单药组(t=37.986、54.622,P<0.05)。与空白对照组相比较,各加药组T-47D细胞Caspase-3、Caspase-8、Bax、Bcl-2蛋白表达差异无显著性(P>0.05)。结论甘露糖能抑制乳癌细胞增殖,并增加乳癌细胞对EPI敏感性,MPI表达较低的乳癌细胞对甘露糖更敏感。

Objective To investigate the effect of mannose on the proliferation of breast cancer cells and their chemosensitivity to epirubicin(EPI).Methods Western blotting was used to measure the expression of mannose phosphate isomerase(MPI),a key protein for mannose metabolism,in breast cancer cells(BT-549,HCC1937,T-47D,and MCF-7)and normal human breast epithelial cells(MCF-10A).MCF-7 and T-47D cells were randomly divided into control group(cultured in a common culture solution containing fetal bovine serum at a volume fraction of 0.10),mannose group(25 mmol/L mannose),glucose group(25 mmol/L glucose),EPI group(1μmol/L EPI),mannose+EPI group(1μmol/L EPI+25 mmol/L mannose),and glucose+EPI group(1μmol/L EPI+25 mmol/L glucose),and the above groups were cultured under both aerobic and anaerobic conditions.MTT colorimetry was used to measure the effect of mannose or mannose combined with EPI on cell growth activity,and Western blotting was used to measure the expression of the apoptosis-related proteins Caspase-3,Caspase-8,Bax,and Bcl-2 in T-47D cells in the control group,the mannose group,and the glucose group.Results There was a significant difference in the expression of MPI between MCF-10A,BT-549,HCC1937,T-47D,and MCF-7 cells(F=392.680,P<0.05).Compared with MCF-10A cells,breast cancer T-47D cells had significantly lower expression of MPI and MCF-7 cells had significantly higher expression of MPI(t=25.449,32.857;P<0.05).Under aerobic conditions,compared with the control group,the mannose group had a significant reduction in the proliferation rate of T-47D cells(t=33.338,P<0.05),while there was no significant difference in the proliferation rate of MCF-7 cells between the two groups(P>0.05);the mannose+EPI group had a significantly lower proliferation rate of T-47D cells than the EPI group(t=22.901,P<0.05),and there was no significant difference in the proliferation rate of MCF-7 cells between the mannose+EPI group and the control group(P>0.05).Under hypoxic conditions,compared with the control group,the mannose group had significant reductions in the proliferation rates of T-47D and MCF-7 cells(t=24.538,35.158;P<0.05),and the mannose+EPI group had significantly lower proliferation rates of T-47D and MCF-7 cells than the EPI group(t=37.986,54.622;P<0.05).There were no significant differences in the protein expression of Caspase-3,Caspase-8,Bax,and Bcl-2 in T-47D cells between the blank control group and the drug treatment groups(P>0.05).Conclusion Mannose can inhibit the proliferation of breast cancer cells and enhance their sensitivity to epirubicin.Breast cancer cells with lower expression of MPI are more sensitive to mannose.