蜂毒肽通过转化生长因子βSMAD信号通路调控非小细胞肺癌的上皮-间质转化进程  

Regulation of melittin on non-small cell lung cancer by TGF-β/SMAD on inhibiting TGF-β-induced EMT process

作　　者：阿布力米提·阿布来提 孙伟[1] 阿迪力·萨来[1] 孙晓宏[1] 瓦热斯江·衣不拉音[1] 高云飞[1] 初建虎[1] 徐克明 ABULIMITI·Abulaiti;SUN Wei;ADILI·Salai;SUN Xiaohong;WARESIJIANG·Yibulayin;GAO Yunfei;CHU Jianhu;XU Keming(Department of Thoracic Surgery, Cancer Hospital Affiliated to Xinjiang Medical University, Urumqi 830011, China)

机构地区：[1]新疆医科大学附属肿瘤医院胸外科,新疆乌鲁木齐830011

出　　处：《西部医学》2022年第1期21-27,共7页Medical Journal of West China

基　　金：中国民族医药学会科研项目(2019KYXM-Z196-20)。

摘　　要：目的探讨蜂毒肽对非小细胞肺癌转化生长因子β(TGF-β)/SMAD信号通路的调控作用及对TGF-β1诱导的上皮-间质转化(EMT)进程的影响。方法通过外源性TGF-β1刺激构建非小细胞肺癌A549细胞EMT模型,将细胞分为对照组、TGF-β1组(加入2μg/L TGF-β1)、蜂毒肽组(加入2.5 mg/L蜂毒肽)、蜂毒肽+TGF-β1组(加入2μg/L TGF-β1和2.5 mg/L蜂毒肽)。采用免疫印迹及逆转录聚合酶链反应(RT-PCR)实验检测各组细胞EMT标志蛋白钙黏附蛋白E、N-钙黏蛋白、波形蛋白及mRNA表达情况;同时检测各组细胞中Smad2、p-Smad2蛋白表达情况,分析蜂毒肽对非小细胞肺癌TGF-β/SMAD信号通路的影响。采用免疫荧光实验检测对照组及蜂毒肽组细胞中TGF-βⅠ受体及TGF-βⅡ受体表达情况,同时采用Transwell实验检测两组细胞迁移、侵袭能力。结果①与对照组相比,TGF-β1组钙黏附蛋白E mRNA及蛋白表达水平明显降低,N-钙黏蛋白mRNA及蛋白与波形蛋白mRNA及蛋白表达水平明显升高(均P<0.05)。蜂毒肽+TGF-β1组钙黏附蛋白EmRNA及蛋白表达水平明显优于TGF-β1组,且N-钙黏蛋白mRNA、蛋白以及波形蛋白mRNA、蛋白表达水平低于TGF-β1组(均P<0.05)。②对照组和蜂毒肽组细胞中TGF-βⅠ受体表达比较,差异无统计学意义(P>0.05),蜂毒肽组中TGF-βⅡ受体含量明显少于对照组。③Transwell实验结果显示,TGF-β1组细胞迁移、侵袭能力明显优于对照组,蜂毒肽+TGF-β1组细胞迁移、侵袭能力明显劣于TGF-β1组细胞(均P<0.05)。结论蜂毒肽可通过抑制TGF-β/SMAD信号通路抑制Smad2磷酸化,下调TGF-βⅡ受体分布,从而抑制肿瘤细胞的侵袭、迁移,抑制TGF-β1诱导的肺癌细胞EMT进程。Objective To investigate the regulatory effect of betulin on transforming growth factor-β/Smad signaling pathway and its effect on TGF-β1-induced epithelial-mesenchymal transition process of non-small cell lung cancer.Methods The EMT model of non-small cell lung cancer A549 cells was established by exogenous TGF-β1 stimulation.The cells were divided into control group,TGF-β1 group(adding 2μg/L TGF-β1),melitotide group(adding 2.5 mg/L melitotide),TGF-β1+melitotide group(adding 2μg/L TGF-β1 and 2.5 mg/L melitotide).Western blotting and reverse transcription-polymerase chain reaction(RT-PCR)were used to detect the expression levels of EMT marker proteins calcadherin E,n-calcadherin,vimentin and their mRNA.Meanwhile,the protein expressions of Smad2 and p-Smad2 in each group were detected to analyze the effect of melitinin on TGF-β/Smad signaling pathway in NSCLC.Using immunofluorescence experiments testing in the control group and TGF-βⅠmelittin group cells receptors and TGF-βⅡreceptor expression,at the same time the two groups of Transwell experiment testing cell migration,invasion ability.Results Compared with the control group,TGF-β1 group significantly decreased the expression levels of cadcadin emRNA and protein,and N-cadcadin mRNA and protein and vimentin mRNA and protein,the expression level was significantly increased(P<0.05).The expression levels of cadherin emRNA and protein in melittin+TGF-β1 group were significantly higher than those in TGF-β1 group.N-cadherin mRNA and protein in melittin+TGF-β1 group were significantly higher than those in TGF-β1 group and vimentin mRNA,protein were lower than TGF-β1 group(<0.05).In the control group and melittin group cells TGF,there was no statistically significant difference betaⅠreceptor expression(P>0.05),TGF-betaⅡreceptor content in melittin group obviously less than control group.Transwell test results showed that the cell migration and invasion ability of TGF-β1 group was significantly better than that of the control group cells and The

关 键 词：蜂毒肽 非小细胞肺癌 转化生长因子β/SMAD 转化生长因子βⅠ/Ⅱ受体 上皮-间质转化 

分 类 号：R-33[医药卫生]