依西美坦联合化疗对乳腺癌模型小鼠基质金属蛋白酶-9及上皮细胞钙黏蛋白表达的影响

Effects of exemestane combined with chemotherapy on the expressions of matrix metalloprotein-9 and E-cadherin in breast cancer mice

目的研究依西美坦联合化疗对乳腺癌模型小鼠基质金属蛋白酶-9(MMP-9)及上皮细胞钙黏蛋白(E-cadherin)表达影响。方法选取SPF级BALB/c雌性小鼠,向小鼠右腋皮下注射4T1小鼠乳腺癌细胞悬液0.5 m L,肿瘤结节≥2 mm×2 mm为造模成功。按照体重将造模成功小鼠随机分成3组(均灌胃给药):内分泌组(8只,依西美坦片25 mg)、化疗组(8只,TP方案:紫杉醇135 mg·m-2+顺铂20 mg·m-2)、联合组(9只,依西美坦片25 mg+TP方案),正常组6只;均给药3周。3周后处死小鼠,用荧光定量PCR、免疫组化及免疫印迹法检测4组小鼠右腋皮下组织MMP-9及E-cadherin表达水平。结果小鼠MMP-9蛋白的相对含量:与正常组的0.01±0.00比较,内分泌组、化疗组与联合组分别为0.75±0.37,0.71±0.25,0.33±0.01,差异均有统计学意义(均P<0.05);与联合组相比,内分泌组和化疗组小鼠MMP-9蛋白相对含量明显升高,差异均有统计学意义(均P<0.01)。小鼠E-cadherin蛋白的相对含量:与正常组的0.64±0.28比较,内分泌组、化疗组与联合组分别为0.17±0.08,0.20±0.17,0.44±0.28,内分泌组与化疗组差异均有统计学意义(均P<0.05);与联合组相比,内分泌组差异有统计学意义(P<0.01)。小鼠MMP-9 mRNA的相对含量:与正常组的0.48±0.04比较,内分泌组、化疗组与联合组分别为10.80±0.04,10.54±0.13,3.48±0.04,差异均有统计学意义(均P<0.05);与联合组相比,内分泌组和化疗组差异均有统计学意义(均P<0.01)。小鼠E-cadherin mRNA的相对含量:与正常组的13.36±0.06比较,内分泌组、化疗组与联合组分别为5.13±0.04,5.25±0.02,11.36±0.06,差异均有统计学意义(均P<0.05);与联合组相比,内分泌组和化疗组差异均有统计学意义(均P<0.01)。结论 MMP-9和E-cadherin与乳腺癌的发生有着密切关系,依西美坦联合化疗能够明显改善MMP-9和E-cadherin水平。

Objective To explore the effects of exemestane combined with chemotherapy on the expressions of matrix metalloprotein-9（ MMP-9） and E-cadherin in breast cancer mice. Methods The SPF BALB/c female mice were selected,25 mice with breast cancer model were successfully established and randomly divided them into 3groups： endocrine group with 8 mice and was given 25 mg of exemestane tablets by gavage once a day; chemotherapy group with 8 mice and was given a range of TP chemotherapy（ paclitaxel,135 mg · m^-2andcisplatinum 20 mg·m^-2）; combined group with 9 mice and was given 25 mg of exemestane tablets by gavage once a day and a range of TP chemotherapy. The normal group contained 6 normal healthy mice and was given the same feeding without drugs. After feeding for 3 weeks,all mice were killed. The expression levels of MMP-9 and E-cadherin in mice right axillary subcutaneous tissue of the four groups were investigated by fluorescence quantitative PCR,immunohistochemistry and Western blot. Results The expression levels of MMP-9 protein in mice： Compared with normal grouP〈0. 01 ± 0. 00,endocrine group,chemotherapy group,combined group were 0. 75 ± 0. 37,0. 71 ± 0. 25,0. 33 ± 0. 01 with increased significantly（ all P〈0. 05）; compared with combined group,endocrine group and chemotherapy group increased significantly（ all P〈0. 01）. The expression levels of E-cadherin protein in mice：Compared with normal grouP〈0. 64 ± 0. 28,endocrine group,chemotherapy group,combined group were 0. 17 ± 0. 08,0. 20 ± 0. 17,0. 44 ± 0. 28 with decreased significantly（ all P〈0. 05）; compared with combined group,endocrine group decreased significantly（ all P〈0. 01）. The expression levels of MMP-9 mRNA（ MMP-9/β-actin） in mice：Compared with normal grouP〈0. 48 ± 0. 04,endocrine group,chemotherapy group,combined group were 10. 80 ± 0. 04,10. 54 ± 0. 13,3. 48 ± 0. 04 with increased significantly（ all P〈0. 05）; compared with combined group,endocrine group decreased significantly（ all P〈0. 01）. The expression levels of E-cadherin mRNA（ E-cadherin/β-actin） in mice： Compared with normal group 13. 36 ± 0. 06,endocrine group,chemotherapy group,combined group were5. 13 ± 0. 04,5. 25 ± 0. 02,11. 36 ± 0. 06 with decreased significantly（ all P〈0. 05）; compared with combined group,endocrine group decreased significantly（ P〈0. 01）. Conclusion There are close relationships between the expression levels of MMP-9 and E-cadherin and breast cancer,and the combination of exemestane and chemotherapy could significantly improve the expression levels of MMP-9 and E-cadherin.