[Objective]To explore the effects of apigenin 7-O-glucoside(AGL)on the apoptosis of anti-tumor necrosis factor-related apoptosis inducing ligand(TRAIL)breast cancer cells.[Methods]The effects of AGL,TRAIL and AGL+TRAIL on activity of breast cancer cells,level of poly(ADP-ribose)polymerase-1(PARP-1)and apoptosis were detected by CCK-8,Western-blot and flow cytometry.ROS level was detected by laser scanning confocal microscope.The level of malondialdehyde(MDA)and activity of superoxide dismutase(SOD)were detected by ELISA.The levels of PARP-1,p53 and p53up-regulated modulator of apoptosis(PUMA)were detected by Western-blot.MCF7 models of transplanted tumor were established to observe growth and apoptosis of transplanted tumor treated with AGL+TRAIL.[Results]With the treatment of TRAIL(50 ng/mL)+AGL(20,40 μmol/L),survival rate of MCF7 cells was decreased(P＜0.05),and Cleaved PARP-1 level was increased(P＜0.05).With the treatment of TRAIL(50 ng/mL)+AGL(20 μmol/L),apoptosis rate of MCF7 cells,levels of Cleaved-Cas-3 and Cleaved-Cas-9 were significantly increased(P＜0.05).With the treatment of AGL(10,20 μmol/L),number of ROS(+)cells and MDA level in MCF7 cells were significantly increased(P＜0.05),while SOD activity was significantly decreased(P＜0.05).The number of ROS(+)cells in MCF7 cells,related levels of p53,PUMA and Cleaved PARP-1 proteins with the treatment of TRAIL(50 ng/mL)+ AGL(20 μmol/L)were significantly higher than those with the treatment of TRAIL(50 ng/mL)(P＜0.05),which were significantly lower with the treatment of TRAIL(50 ng/mL)+AGL(20 μmol/L)+NAC(4 mmol/L)than those with the treatment of TRAIL(50 ng/mL)+AGL(20 μmol/L)(P＜0.05).The mass and volume of transplanted tumors,positive rates of p53 and PUMA3 in AGL+TRAIL group were significantly lower than those in Control group(P＜0.05),while apoptosis rate was significantly higher than that in Control group(P＜0.05).[Conclusion]AGL combined with TRAIL can promote apoptosis of anti-TRAIL MCF7 cells,which may be related to enhancing oxidative stress damage and activating caspase cascade.
目的 探讨微小核糖核酸-205-5p(miR-205-5p)靶向真核细胞翻译起始因子4E(eIF4E)对人乳腺癌细胞MCF-7增殖、侵袭及上皮间质转化的影响,为乳腺癌(MC)新型靶向治疗药物的研发提供参考.方法 对MCF-7进行培养,miR-205-5p转染mimic-NC及miR-205-5p mimic以验证miR-205-5p与eIF4E靶向关系,同时将培养好的MCF-7细胞分为对照组(正常培养的MCF-7细胞)、mimic-NC组、pcDNA组、miR-205-5p mimic组、miR-205-5p组、eIF4E-pcDNA组、miR-205-5p+eIF4E-pcDNA组,分析miR-205-5p靶向eIF4E对MCF-7细胞增殖、迁移和上皮间质转化的影响.结果 miR-205-5p mimic组的miR-205-5p相对表达量显著高于对照组和mimic-NC组,而eIF4E的mRNA表达水平则显著低于对照组和mimic-NC组(P<0.05),而eIF4E野生型、mimic均为阳性的Lucifera-se activity(R/F ratio)更低.eIF4E-pcDNA组eIF4E蛋白和mRNA相对表达量较对照组、pcDNA组明显高(P<0.05).miR-205-5p组MCF-7细胞阳性率、菌落分布率和Ki67、PCNA蛋白表达水平较对照组、eIF4E-pcDNA组、miR-205-5p+eIF4E-pcDNA组明显低(P<0.05).miR-205-5p组侵袭性细胞阳性率和侵袭性细胞数量较对照组、miR-205-5p+eIF4E-pcDNA组、eIF4E-pcDNA组明显低(P<0.05),而eIF4E-pcDNA组侵袭性细胞数量最高.miR-205-5p组MCF-7细胞上皮间质标志物E-Cadherin、N-Cadherin、Vimentin阳性表达率和蛋白表达水平明显低于对照组、miR-205-5p+eIF4E-pcDNA组、eIF4E-pcDNA组(P<0.05),而eIF4E-pcDNA组上皮间质标志物E-Cadherin、N-Cadherin、Vimentin阳性表达率和蛋白表达水平显著高于其他组(P<0.05).结论 miR-205-5p负靶向eIF4E能有效抑制MCF-7增殖、侵袭及上皮间质转化,有望为MC新型基因靶向治疗药物的研发提供重要参考.
目的 探究miR-10b-3p对乳腺癌细胞增殖、迁移和侵袭的影响及其潜在机制.方法 下载GEO数据、TCGA_BRCA乳腺癌数据矩阵和临床信息,分析miR-10b-3p、抑制素βA(INHBA)mRNA表达水平与乳腺癌的关系;构建稳定过表达miR-10b-3p的MCF-7和MDA-MB-231细胞系,CCK-8、划痕和Transwell小室实验检测乳腺癌细胞增殖、迁移与侵袭能力,体外构建裸鼠荷瘤模型,观察过表达miR-10b-3p对瘤体生长的影响,双荧光素酶报告基因检测miR-10b-3p与INHBA的靶向关系.结果 GEO数据集中,与癌旁正常组织比较,乳腺癌组织中miR-10b-3p水平降低(P<0.05);miR-10b-3p水平在不同临床分期、T分期及临床组织病理学G分级患者中存在差异(P<0.05);TCGA数据库中,与癌旁对照组织比较,乳腺癌组织中INHBA mRNA水平升高(P<0.05);INHBA mRNA水平在不同临床分期、N分期、预后情况患者中存在差异(P<0.05);miR-10b-3p与INHBA存在靶向关系,与miR-NC组比较,miR-10b-3p组MCF-7、MDA-MB-231细胞中miR-10b-3p水平上调,INHBA表达、细胞增殖能力、迁移率及侵袭能力均降低(P<0.05),INHBA可逆转上述作用;与miR-NC组比较,miR-10b-3p组裸鼠瘤体体积、质量均降低,miR-10b-3p水平升高(P<0.05).结论 miR-10b-3p可能靶向调控INHBA,抑制乳腺癌细胞增殖、迁移和侵袭.
