Objective To explore the effect of vascular endothelial growth factor-C (VEGF-C) gene transfection on the expression level of VEGF-C in human breast cancer MCF-7 cell. Methods The constructed VEGF-C gene eukaryotic expression vector was transfected into the human breast cancer MCF-7 cell by using lipofectamine transfection reagents， and the positive cell clones were obtained through G418 selection after transfection. The expressions of VEGF-C mRNA and protein were detected by RT-PCR and Western blot respectively. Results Following the transfection of the VEGF-C recombination plasmid, there were significant differences on the expression levels of VEGF-C mRNA and protein between pcDNA3.1-VEGF-C transfection group and pcDNA3.1 transfection group （12.382±2.183 vs 6.039±1.950, P＜0.01; 0.971±0.186 vs 0.594±0.196, P＜0.05). Conclusion With the transfection of pcDNA3.1-VEGF-C vector by using the liposome, the expression levels of VEGF-C mRNA and protein rise up in breast cancer MCF-7 cell.
ObjectiveTo explore expression of Mdm2 in the estrogen receptor α (ERα)-positive breast cancer tissues and fibroadenoma of breast tissues, and to explore the effect of MDM2-siRNA on cell proliferation, colony formation, and apoptosis for MCF-7 cells. Methods① Seventy eight ERα-positive breast cancer patients identified by histopathological examination, who underwent surgery in our hospital from June 2012 to October 2015, as well as 10 fibroadenoma of breast patients underwent surgery in the same period, were collected retrospectively to determine the expression of Mdm2, then explore the relationship between the expression of Mdm2 and clinical pathological characteristics of ERα-positive breast cancer patients. ② MCF-7 cells were divided to MDM2-siRNA group (added with MDM2-siRNA), negative control group (added with negative siRNA), and blank control group (added without any reagent). Expression of Mdm2, cell proliferation rate, number of colony formation, and apoptosis rate were determined in the MCF-7 cells of 3 groups. Results① No one of fibroadenoma of breast patients was found positive expression of Mdm2 (0/10), and 38 of 78 ERα-positive breast cancer patients were found the positive expression of Mdm2 (48.7%), which is higher than that of fibroadenoma of breast tissues (χ2=12.357, P=0.000). In ERα-positive breast cancer patients, expression of Mdm2 was related with TNM staging and number of metastasic lymph node (P < 0.050), the positive expression rate of Mdm2 was higher in patients with later TNM staging or more metastasic lymph node. ② Cell proliferation rates on 2, 3, and 4 days after transfection, expression level of Mdm2, and number of colony formation were all lower (P < 0.050), but the apoptosis rate was higher in MDM2-siRNA group (P < 0.050), comparing with negative control group and blank control group. But there was no significant difference between negative control group and blank control group on aforementioned indexes (P > 0.050). ConclusionMdm2 is a diagnostic marker in ERα-positive breast cancer patients, and treatment targeting it might has a certain therapeutic value.
目的:探讨雌激素饥饿对肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导乳腺癌细胞MCF-7凋亡的影响及作用机制。〖HTH〗方法〖HTSS〗:MCF-7细胞培养贴壁之后,用雌激素饥饿处理后加入TRAIL,用荧光染料Hoechst染色法检测细胞的凋亡,用细胞计数法检测细胞的存活。在雌激素饥饿处理MCF-7细胞后,收集对照和饥饿组蛋白用Western blot法检测相关的蛋白表达。〖HTH〗结果〖HTSS〗:单独使用雌激素饥饿处理或者单独使用TRAIL处理乳腺癌细胞MCF-7都能诱导细胞凋亡,但是它们诱导凋亡的活性较小,两种方法联合使用可以极大地增加诱导细胞凋亡的活性(Plt;0.001)。雌激素饥饿处理后的乳腺癌细胞,死亡受体5(DR5)表达上调。〖HTH〗结论〖HTSS〗:乳腺癌细胞MCF-7对TRAIL敏感度不高,雌激素饥饿可以增加TRAIL诱导乳腺癌细胞凋亡的活性,DR5与雌激素饥饿诱导TRAIL活性增加相关。