三阴性乳腺癌(triple negative breast cancer,TNBC)恶性程度高,预后差,其p53基因的突变频率达80%以上。日蟾蜍它灵(gamabufotalin,CS-6)是日本蟾蜍的皮肤分泌物,有研究表明CS-6具有抗癌作用,但其对TNBC的作用和机制尚无报道。本文采用...三阴性乳腺癌(triple negative breast cancer,TNBC)恶性程度高,预后差,其p53基因的突变频率达80%以上。日蟾蜍它灵(gamabufotalin,CS-6)是日本蟾蜍的皮肤分泌物,有研究表明CS-6具有抗癌作用,但其对TNBC的作用和机制尚无报道。本文采用MTT法和细胞克隆形成实验检测了不同浓度CS-6对TNBC细胞系MDA-MB-468和MDA-MB-231细胞增殖的影响;采用Western-blot检测了CS-6在外源和内源水平对p53和热休克蛋白90(heat shock protein 90,HSP90)表达水平的影响;采用联合指数及等效线分析法考察了CS-6和HSP90的抑制剂坦螺旋霉素(tanespimycin,17-AAG)对MDA-MB-468和MDA-MB-231细胞增殖的协同作用。结果显示,CS-6明显抑制MDA-MB-468和MDA-MB-231细胞的增殖,其抑制增殖机制有可能与降解HSP90/突变p53有关;而且,CS-6和HSP90抑制剂17-AAG联合用药对抑制MDA-MB-468和MDA-MB-231细胞增殖具有协同作用。展开更多
To study the functions of the early auxin-responsive genes Cs GH3.1 and Cs GH3.6 in citrus resistance against canker disease, we cloned Cs GH3.1and Cs GH3.6 in ‘Newhall' Navel Orange(Citrus sinensis Osbeck). They...To study the functions of the early auxin-responsive genes Cs GH3.1 and Cs GH3.6 in citrus resistance against canker disease, we cloned Cs GH3.1and Cs GH3.6 in ‘Newhall' Navel Orange(Citrus sinensis Osbeck). They are 1 797 bp and 1 887 bp and encode 598 and 629 amino acids, respectively. In vitro mature leaves from susceptible ‘Newhall' and resistant Calamondin(C. madurensis) were inoculated by a Xanthomonas axonopodis pv. citri(Xac) bacterial suspension, and expression of Cs GH3.1 and Cs GH3.6 in the two varieties were analyzed using quantitative real-time PCR(q RT-PCR). ‘Newhall' leaves were treated with different hormones for 3 days, inoculated by Xac bacterial suspension, and then the symptoms in these leaves were investigated. We used q RT-PCR to analyze the effect of different hormones on Cs GH3.1 and Cs GH3.6 expression in ‘Newhall'leaves. The expression levels of both Cs GH3.1 and Cs GH3.6 were significantly induced by Xac in ‘Newhall' leaves, compared with levels in Calamondin leaves. 1-naphthy acetic acid(NAA) increased the hypertrophy of infection sites in ‘Newhall' leaves, while naphthyl-phthalamic acid(NPA)had no visible effect on lesion development. NAA hormone greatly improved expression of Cs GH3.1 in ‘Newhall', but not Cs GH3.6. These results indicate that the auxin primary-response gene Cs GH3.1 plays an important role in citrus susceptibility to Xac.展开更多
文摘三阴性乳腺癌(triple negative breast cancer,TNBC)恶性程度高,预后差,其p53基因的突变频率达80%以上。日蟾蜍它灵(gamabufotalin,CS-6)是日本蟾蜍的皮肤分泌物,有研究表明CS-6具有抗癌作用,但其对TNBC的作用和机制尚无报道。本文采用MTT法和细胞克隆形成实验检测了不同浓度CS-6对TNBC细胞系MDA-MB-468和MDA-MB-231细胞增殖的影响;采用Western-blot检测了CS-6在外源和内源水平对p53和热休克蛋白90(heat shock protein 90,HSP90)表达水平的影响;采用联合指数及等效线分析法考察了CS-6和HSP90的抑制剂坦螺旋霉素(tanespimycin,17-AAG)对MDA-MB-468和MDA-MB-231细胞增殖的协同作用。结果显示,CS-6明显抑制MDA-MB-468和MDA-MB-231细胞的增殖,其抑制增殖机制有可能与降解HSP90/突变p53有关;而且,CS-6和HSP90抑制剂17-AAG联合用药对抑制MDA-MB-468和MDA-MB-231细胞增殖具有协同作用。
基金supported by the China Agriculture Research System(CARS-27)the Fundamental Research Funds for the Central Universities(XDJK2014A018)+1 种基金National Natural Science Foundation of China(31272150)Key Projects in the National Science & Technology Pillar Program during the Twelfth Five-year Plan Period(2012BAD19B6)
文摘To study the functions of the early auxin-responsive genes Cs GH3.1 and Cs GH3.6 in citrus resistance against canker disease, we cloned Cs GH3.1and Cs GH3.6 in ‘Newhall' Navel Orange(Citrus sinensis Osbeck). They are 1 797 bp and 1 887 bp and encode 598 and 629 amino acids, respectively. In vitro mature leaves from susceptible ‘Newhall' and resistant Calamondin(C. madurensis) were inoculated by a Xanthomonas axonopodis pv. citri(Xac) bacterial suspension, and expression of Cs GH3.1 and Cs GH3.6 in the two varieties were analyzed using quantitative real-time PCR(q RT-PCR). ‘Newhall' leaves were treated with different hormones for 3 days, inoculated by Xac bacterial suspension, and then the symptoms in these leaves were investigated. We used q RT-PCR to analyze the effect of different hormones on Cs GH3.1 and Cs GH3.6 expression in ‘Newhall'leaves. The expression levels of both Cs GH3.1 and Cs GH3.6 were significantly induced by Xac in ‘Newhall' leaves, compared with levels in Calamondin leaves. 1-naphthy acetic acid(NAA) increased the hypertrophy of infection sites in ‘Newhall' leaves, while naphthyl-phthalamic acid(NPA)had no visible effect on lesion development. NAA hormone greatly improved expression of Cs GH3.1 in ‘Newhall', but not Cs GH3.6. These results indicate that the auxin primary-response gene Cs GH3.1 plays an important role in citrus susceptibility to Xac.
