miR-423-5p调控TGF-β/Smad信号通路对肺癌脑转移模型大鼠的干预作用探究

The intervention effect of miR-423-5p regulating TGF-β/Smad signal pathway on lung cancer brain metastasis model rats

目的探究miR-423-5p调控TGF-β/Smad信号通路对肺癌脑转移模型大鼠的干预作用。方法2019年2月—2020年1月于同济大学附属东方医院转化医学研究中心进行实验。选取SPF级大鼠60只,随机数字表法分为空白组、模型组、上调组、下调组各15只,除空白组外,其余大鼠建立肺癌脑转移模型,建模成功后,上调组、下调组大鼠分别静脉注射miR-423-5p mimics control、miR-423-5p mimics,空白组、模型组大鼠静脉注射等体积生理盐水。干预14 d后比较各组大鼠肺组织病理组织学变化,miR-423-5p表达量,血清癌胚抗原(CEA)、鳞状细胞癌相关抗原(SCC-Ag)、细胞角蛋白19血清片段21-1(CYFRA21-1)水平,肺泡灌洗液中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、IL-6水平及TGF-β/Smad信号通路蛋白表达量。结果与空白组比较,模型组miR-423-5p表达量、CEA、SCC-Ag、CYFRA21-1、TNF-α、IL-1β、IL-6水平及TGF-β1、COl-1、Smad2/3、Vimentin蛋白表达量均升高(P均<0.01);与模型组比较,上调组上述指标水平升高,下调组降低(P均<0.01);与上调组比较,下调组上述指标水平均降低(t/P=9.513/0.001、5.936/0.001、5.819/0.001、4.833/0.001、4.465/0.001、7.416/0.001、4.114/0.001、7.320/0.001、6.479/0.001、5.504/0.001、4.880/0.001)。结论下调miR-423-5p可能通过作用于TGF-β/Smad信号通路,调控TGF-β1、COl-1、Smad2/3、Vimentin蛋白表达量,阻断炎性反应,抑制肺癌脑转移的发生发展。

Objective To explore the intervention effect of miR-423-5p regulating TGF-β/Smad signaling pathway on lung cancer brain metastasis model rats.Methods From February 2019 to January 2020,the experiment was carried out in the Translational medicine research center of Translational Medicine Research Center,Dongfang Hospital,Tongji University.Sixty SPF Rats were selected,including 15 as blank group.The rest rats were randomly divided into model group,up-regulation group and down-regulation group with 15 rats in each group.After successful modeling,rats in up-regulation group and down-regulation group were injected with miR-423-5p mimics control and miR-423-5p respectively Mice in blank group and model group were injected with equal volume of normal saline.Histopathological changes,expression of miR-423-5p,serum carcinoembryonic antigen(CEA),squamous cell carcinoma associated antigen(SCC Ag),cytokeratin 19 serum fragment 21-1(CYFRA21-1),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)in alveolar lavage fluid(BALF),and TGF-β/Smad signaling pathway were compared Protein expression.Results Compared with the up-regulation group,the expression of miR-423-5pp in the down-regulation group was lower(t/P=9.513/0.001);compared with the up-regulation group,the expression of miR-423-5pp in the down-down group was lower(t/P=5.936/0.001,5.819/0.001,4.833/0.001);compared with the up-regulation group,the expression of TNF-α,IL-1β,IL-6 IL-6 in the down-down group was lower(t/P=4.465/0.001,7.416/0.001);compared with the up-up group,the levels of TNF-α,IL-1(t/P=4.114/0.001,7.320/0.001)Compared with the control group,the expression of TGF-β1,COL-1,Smad2/3 and vimentin in the down-regulation group was lower(t/P=6.479/0.001,5.504/0.001,4.880/0.001).Conclusion Down regulation of miR-423-5p may regulate the expression of TGF-β1,COL-1,Smad2/3 and vimentin protein through TGF-β/Smad signaling pathway and block inflammatory response.