塞来昔布联合p65miRNA抑制人乳腺癌移植瘤的生长

Effects of celecoxib combined with p65miRNA on tumor growth in breast cancer xenografts in nude mice

目的探讨下调核因子NF-κB p65亚基的表达联合塞来昔布对人乳腺癌裸鼠移植瘤生长的协同抑制效应及其机制。方法建立人乳腺癌裸鼠移植瘤模型,随机将裸鼠分为6组,control组、Neg-miRNA组、p65miRNA组、塞来昔布组、塞来昔布+Neg-miRNA组、塞来昔布+p65miRNA组。观察治疗前后裸鼠的一般状态、肿瘤体积和瘤重,计算平均抑瘤率。免疫组化法检测p65miRNA干扰后肿瘤组织中p65蛋白的表达。RT-PCR、Western印迹法检测肿瘤组织中血管内皮生长因子(VEGF)、金属基质蛋白酶(MMP-2)基因mRNA及蛋白表达的变化。结果塞来昔布组、p65miRNA组和塞来昔布+p65miRNA组肿瘤生长受到明显抑制,抑瘤率分别为43.23%、40.72%及61.69%。p65miRNA组较对照组p65蛋白表达明显减少。塞来昔布组、p65miRNA组和塞来昔布+p65miRNA组VEGF、MMP-2 mRNA及蛋白表达受到不同程度地抑制。结论塞来昔布及p65miRNA均具有明显的抗肿瘤作用,联合应用时具有一定的协同作用,可显著抑制人乳腺癌裸鼠移植瘤的生长,并可显著下调VEGF、MMP-2的表达。

Objective To evaluate the effects of celecoxib combined with p65miRNA on tumor growth in breast cancer xenografts in nude mice and its mechanism.Methods After successfully establishing breast cancer xenografts in nude mice,the mice were randomly divided into control,the neg-miRNA,the p65 miRNA,the celecoxib,the celecoxib combined with neg-miRNA and the celecoxib combined with p65miRNA groups.On the 42nd day after treatment,tumor volume,weight and the rate of tumor inhibition were measured.Tumor tissues were collected and assessed for the detection of p65,VEGF and MMP-2 by immunohistochemistry,Western blotting and RT-PCR.Results The rate of tumor inhibition was 43.23% in the celecoxib group,40.72% in the p65miRNA group and 61.69% in the celecoxib combined with p65miRNA group.Immunohistochemical results showed a significant decrease in p65 expression of the p65miRNA group.Western blotting and RT-PCR analysis showed that the expressions of VEGF and MMP-2 were significantly reduced in the celecoxib,the p65miRNA and the celecoxib combined with p65miRNA groups.Conclusions The combination of celecoxib and p65miRNA can enhance the anti-tumor effect in human breast cancer xenografts,by down-regulating expressions of VEGF and MMP-2.