白头翁皂苷A通过调控miR-24-3p/RNF2的表达影响乳腺癌细胞增殖及放射敏感性

目的:探讨白头翁皂苷A对乳腺癌细胞增殖及放射敏感性的影响及其作用机制。方法:用浓度为0、5、10、15和20 mg/L的白头翁皂苷A作用于人乳腺癌MCF-7细胞,并转染微小RNA-24-3p(miR-24-3p)过表达载体或抑制表达载体。用MTT法检测细胞活力的变化;集落形成实验检测细胞放射敏感性;RT-qPCR分析miR-24-3p和环指蛋白2(RNF2)的mRNA表达水平;Western blot检测RNF2的蛋白表达;萤光素酶报告基因实验检测miR-24-3p和RNF2的靶向关系。结果:与对照组(0 mg/L)相比,5、10、15和20 mg/L的白头翁皂苷A组MCF-7细胞的增殖抑制率显著升高(P<0.05),白头翁皂苷A处理的MCF-7细胞经射线照射后存活分数显著降低,RNF2表达水平显著降低(P<0.05);miR-24-3p靶向负调控RNF2。白头翁皂苷A和miR-24-3p同时处理的MCF-7细胞经射线照射后,存活曲线下移;抑制miR-24-3p表达可逆转白头翁皂苷A对MCF-7细胞的增殖抑制和放射增敏作用。结论:白头翁皂苷A可能通过miR-24-3p/RN...

Pulsatilla saponin A affects proliferation and radiosensitivity of breast cancer cells by regulating miR-24-3p/RNF2 expression

AIM:To investigate the effect of Pulsatilla saponin A on proliferation and radiosensitivity of breast cancer cells and its mechanism.METHODS:Human breast cancer MCF-7 cells were treated with Pulsatilla saponin A at concentrations of 0,5,10,15 and 20 mg/L and transfected with microRNA-24-3 p(miR-24-3 p)over-expression vector or inhibitory expression vector.The proliferation and radiosensitivity of the MCF-7 cells were measured by MTT assay and colony formation assay.The miR-24-3 p expression and ring finger protein 2(RNF2)mRNA level were detected by RT-qPCR.The protein expression of RNF2 was determined by Western blot.The luciferase reporter assay was used to detect the targeting relationship between miR-24-3 p and RNF2.RESULTS:Compared with control group(0 mg/L),the proliferation inhibitory rate of the MCF-7 cells was significantly increased in 5,10,15 and 20 mg/L Pulsatilla saponin A groups(P<0.05).The survival score of the MCF-7 cells treated with Pulsatilla saponin A was significantly decreased after irradiation,and the expression of RNF2 was significantly decreased(P<0.05).miR-24-3 p targeted RNF2 and negatively regulated its expression.When the MCF-7 cells were simultaneously treated with Pulsatilla saponin A and miR-24-3 p,the cell survival curve significantly shifted down.Inhibition of miR-24-3 p expression reversed the proliferation-inhibiting and radiation-sensitizing effects of Pulsatilla saponin A on the MCF-7 cells.CONCLUSION:Pulsatillasaponin A may affect the proliferation and radiosensitivity of breast cancer cells through miR-24-3 p/RNF2 signaling pathway.