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目的探讨青藤碱对人乳腺癌MCF-7细胞系增殖的抑制作用及其对环氧合酶-2(COX-2)和血管内皮生长因子(VEGF)表达的影响。方法以四甲基偶氮唑蓝(MTT)法观察不同浓度青藤碱(0.1、0.25、0.5、1.0、2.5、5.0 mmol/L)对人乳腺癌MCF-7细胞增殖的影响;以实时聚合酶链反应(real-time PCR)分析青藤碱(0.1、0.25、0.5、1.0、2.5、5.0 mmol/L)对人乳腺癌MCF-7细胞COX-2和VEGF mRNA表达的影响;以酶联免疫(ELISA)方法分析青藤碱(0.5、1.0、2.5 mmol/L)对人乳腺癌MCF-7细胞上清VEGF含量的影响。结果 (1)将对照组定义为100%,青藤碱浓度在0.25、0.5、1.0、2.5、5.0 mmol/L时,出现剂量依赖性的MCF-7细胞增殖率降低(78%、68%、53%、35%、17%,P均<0.05)。(2)将对照组定义为1,0.25、1.0、2.5 mmol/L青藤碱组使MCF-7细胞COX-2mRNA相对表达值依次递降至0.75±0.12、0.64±0.15、0.35±0.14;VEGF mRNA相对表达值依次递降低为0.88±0.08、0.67±0.13、0.46±0.11,差异均有统计学意义(P均<0.05)。(3)MCF-7细胞上清VEGF含量按对照组、青藤碱0.25、1.0、2.5 mmol/L组之序[(90.3±8.2)、(76.6±9.7)、(55.7±7.2)、(31.2±5.2)pg/ml]依次降低(P<0.05)。结论青藤碱能够抑制人乳腺癌MCF-7细胞的增殖,其作用机制可能是通过抑制COX-2表达,从而影响VEGF的表达和释放。
Objective To investigate the inhibitory effect of sinomenine on the proliferation of human breast cancer cell line MCF-7 and its effect on the expression of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF). Methods The effects of different concentrations of sinomenine (0.1, 0.25, 0.5, 1.0, 2.5 and 5.0 mmol / L) on the proliferation of human breast cancer cell line MCF-7 were observed by MTT assay. The effects of sinomenine (0.1,0.25,0.5,1.0,2.5,5.0 mmol / L) on the mRNA expression of COX-2 and VEGF in human breast cancer MCF-7 cells were analyzed by real-time PCR. The effect of sinomenine (0.5,1.0,2.5 mmol / L) on VEGF in supernatant of human breast cancer MCF-7 cells was analyzed by ELISA. Results (1) When the control group was defined as 100%, the dose-dependent proliferation rate of MCF-7 cells was decreased when the concentration of sinomenine was 0.25,0.5,1.0,2.5 and 5.0 mmol / L (78%, 68% 53%, 35%, 17%, P <0.05). (2) The control group was defined as 1, 0.25, 1.0, 2.5 mmol / L sinomenine group so that the relative expression of COX-2 mRNA in MCF-7 cells decreased in turn down to 0.75 ± 0.12,0.64 ± 0.15,0.35 ± 0.14; VEGF mRNA The relative expression value decreased in turn was 0.88 ± 0.08,0.67 ± 0.13,0.46 ± 0.11, the difference was statistically significant (P all <0.05). (3) The content of VEGF in the supernatant of MCF-7 cells was controlled by the order of (90.3 ± 8.2), (76.6 ± 9.7), (55.7 ± 7.2), (31.2 ± 5.2) pg / ml] (P <0.05). Conclusion Sinomenine can inhibit the proliferation of human breast cancer MCF-7 cells, and its mechanism may be through inhibiting the expression of COX-2, thereby affecting the expression and release of VEGF.