Notch3增强三阴性乳腺癌MDA-MB-231细胞对顺铂的敏感性

Overexpression of Notch3enhance the efficacy of cisplatin to MDA-MB-231

目的:探讨干细胞相关因子Notch3在三阴性乳腺癌MDA-MB-231细胞对顺铂化疗敏感性中的作用。方法:用Lipofectamine2000试剂转染乳腺癌MDA-MB-231细胞并用G418筛选稳定转染细胞系,根据转染质粒不同分为MDA-MB-231-PCLE组(对照组)和MDA-MB-231-N3ICD组(过表达Notch3组),应用实时荧光定量PCR和Western blot方法检测不同组间Notch3表达的差异;用CCK8法检测各组MDA-MB-231细胞对顺铂敏感性的差异;同时利用Lipofectamine 2000转染Notch3的小干扰序列,根据转染序列不同分为T47D-NC(对照组)和T47D-si Notch3(Notch3敲减组);采用免疫印迹检测Notch3表达改变后凋亡相关因子caspase-3的表达情况。结果:与对照组比较,高表达Notch3后三阴性乳腺癌对顺铂的敏感性增加(P<0.05),同时caspase-3的表达随着Notch3的增加而增加,而在高表达Notch3的乳腺癌T47D细胞中,敲减Notch3的表达使caspase-3表达减少。结论:高表达Notch3可以增加三阴性乳腺癌MDA-MB-231细胞对顺铂的敏感性,从而增强患者化疗疗效。

OBJECTIVE： To investigate the effect of stem cell-related factor,Notch3 on human triple negative breast cancer cell,MDA-MB-231 and its resistance to cisplatin,and explore if overexpressed Notch3 can enhance the sensitivity of resistance cells to cisplatin. METHODS：Lipofectamine 2000 was applied to transfect Notch3 plasmid into MDA-MB-231. These cells were divided into MDA-MB-231-PCLE group and MDA-MB-231-N3 ICD group,according to the use of different plasmids. RT-PCR and Western blot methods were used to analyze the expression of Notch3 in different groups. The effects of cisplatin on cell proliferation were assessed by CCK8 assay and evaluated by Graph Pad Prism 5. Lipofectamine 2000 was also applied to transfect the si Notch3 or scramble to cells and they were divided into T47D-NC and T47D-si Notch3 groups. The protein levels of apoptosis-related factor were detected using RT-PCR methods and Western blot. RESULTS：Then efficacy of cisplatin to triple negative breast cancer cells was enhanced by overexpression of Notch3. Apoptosis-related factor,caspase-3 was up-regulated by Notch3 overexpression. However, si Notch3 induced the downregulation of caspase-3 in T47 D,in which the expression of Notch3 was high. CONCLUSION：Overexpression of Notch3 enhanced the sensitivity of MDA-MB-231 to cisplatin which suggests that this m ay improve the efficiency of chemotherapy in patients with breast cancer.