摘要
目的:探讨扶正祛毒方对人乳腺癌MCF-7细胞增殖的抑制作用及对PI3K/Akt信号通路和血管内皮生长因子C(VEGF-C)、血管内皮细胞生长因子受体3(VEGFR-3)表达的影响。方法:人乳腺癌细胞株MCF-7经完全培养基于37℃、5%CO2、完全饱和湿度培养箱中培养至对数生长周期,接种于96孔培养板,加入浓度梯度(5、10及20μmol/L)扶正祛毒方培养基,以正常培养细胞作为对照;孵育24 h后采用噻唑蓝法检测细胞增殖情况,计算细胞增殖抑制率;采用逆转录聚合酶链反应法检测VEGF-C、VEGFR-3 mRNA水平;采用蛋白质印迹法检测PI3K、AKT蛋白表达水平。结果:扶正祛毒方低、中及高剂量组细胞增殖抑制率明显高于对照组,差异均有统计学意义(P<0.05),提示扶正祛毒方对乳腺癌细胞增殖具有较好的抑制效果,且随着剂量的增加抑制效果越明显;扶正祛毒方低、中及高剂量组细胞VEGF-C、VEGFR-3 mRNA水平明显低于对照组,差异均有统计学意义(P<0.05),提示扶正祛毒方能够下调VEGF-C、VEGFR-3基因水平,且随着剂量的增加下调幅度越大;扶正祛毒方低、中及高剂量组细胞PI3K、Akt蛋白表达水平明显低于对照组,差异均有统计学意义(P<0.05),且呈剂量依赖性,剂量越大降低幅度越明显。结论:扶正祛毒方能够抑制乳腺癌MCF-7细胞增殖,其机制可能与阻断PI3K/Akt信号通路,下调VEGF-C、VEGFR-3基因表达有关。
OBJECTIVE:To probe into the inhibition of Fuzheng Qudu prescription on the human breast cancer MCF-7 cell proliferation and its effects on PI3K/Akt signaling pathway and the expression of vascular endothelial growth factor-C(VEGF-C)and vascular endothelial growth factor receptor-3(VEGFR-3).METHODS:The human breast cancer cell strain MCF-7 was cultured in the incubator of completely medium at 37℃,5%CO2,fully saturated humidity to a logarithmic growth cycle,inoculated in a 96-well culture plate,added with concentration gradients(5,10 and 20μmol/L)of Fuzheng Qudu prescription medium,set normal cultured cells as contrast;after 24 hours of incubation,the thiazole blue method was used to detect cell proliferation and calculate the cell proliferation inhibition rate;the reverse transcription polymerase chain reaction method was used to detect the mRNA levels of VEGF-C and VEGFR-3;Western Blot was used to detect the expression levels of PI3K and AKT protein.RESULTS:The inhibition rate of cell proliferation in the low,medium and high dose groups of Fuzheng Qudu prescription was significantly higher than that of the control group,with statistically significant difference(P<0.05),which suggested that Fuzheng Qudu prescription had a good inhibitory effect on breast cancer cell proliferation,and as the dose increase,the inhibitory effect became more obvious;the mRNA levels of VEGF-C and VEGFR-3 in the low,medium and high dose groups of the Fuzheng Qudu prescription were significantly lower than those of the control group,with statistically significant differences(P<0.05),which suggested that Fuzheng Qudu prescription can down-regulate the VEGF-C and VEGFR-3 genes level,and as the dose increase,the decrease became greater;the expression levels of PI3K and Akt protein in the low,medium and high-dose groups of Fuzheng Qudu prescription were significantly lower than those of the control group,with statistically significant differences(P<0.05),which presented a dose-dependent,and the larger the dose,the more obvious the reduction.CONCLUSIONS:Fuzheng Qudu prescription can inhibit the cell proliferation of breast cancer MCF-7,its mechanism may be related to blocking PI3K/Akt signaling pathway and down-regulating the expression of VEGF-C and VEGFR-3 genes.
作者
时华
杨中
李颖
SHI Hua;YANG Zhong;LI Ying(Dept.of General Surgery,Yanqing Hospital of Beijing Hospital of Traditional Chinese Medicine,Beijing 102100,China;Dept.of Oncology,Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University,Beijing 100010,China)
出处
《中国医院用药评价与分析》
2020年第12期1437-1441,共5页
Evaluation and Analysis of Drug-use in Hospitals of China
基金
首都卫生医学发展科研专项基金(No.201907290018)。
