Treg细胞通过上调TGF-β1和B7-H3表达促进食管癌细胞增殖、迁移和侵袭  

Treg cells promote the proliferation,migration and invasion of esophageal cancer cells by up-regulating the expression of TGF-β1 and B7-H3

作　　者：施我大[1] 张亚军[1] 施展[1] 吴纪祥[1] 常绘文[1] 易忠权 梁晓东[3] 周晶晶 宋建祥[1] Shi Woda;Zhang Yajun;Shi Zhan;Wu Jixiang;Chang Huiwen;Yi Zhongquan;Liang Xiaodong;Zhou Jingjing;Song Jianxiang(Department of Cardiothoracic Surgery,the Sixth Affiliated Hospital of Nantong University,Yancheng Third People's Hospital,Yancheng 224000,China;Central Laboratory,the Sixth Affiliated Hospital of Nantong University,Yancheng Third People's Hospital,Yancheng 224000,China;Department of Pathology,the Sixth Affiliated Hospital of Nantong University,Yancheng Third People's Hospital,Yancheng 224000,China;College of Pharmacy,North China University of Science and Technology,Tangshan 063203,China)

机构地区：[1]江苏省盐城市第三人民医院(南通大学第六附属医院)心胸外科,盐城224000 [2]江苏省盐城市第三人民医院(南通大学第六附属医院)中心实验室,盐城224000 [3]江苏省盐城市第三人民医院(南通大学第六附属医院)病理科,盐城224000 [4]河北省唐山市华北理工大学药学院,唐山063203

出　　处：《中华细胞与干细胞杂志（电子版）》2023年第2期65-75,共11页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：2021年江苏省卫生健委科研项目(M2021089)。

摘　　要：目的探讨调节性T(Treg)细胞对食管癌细胞增殖、迁移和侵袭的影响及分子机制。方法使用流式细胞仪分离患者外周血中的Treg细胞,转化生长因子β1(TGF-β1)、TGF-β1/Smad抑制剂(SB431542)、TGF-β1过表达(TGF-β1 OE)和B7同源物3(B7-H3)干扰(sh-B7-H3)慢病毒载体用于调控食管癌细胞TGF-β1和B7-H3的表达,qRT-PCR和Western blot检测TGF-β1和B7-H3表达水平,CCK-8和Transwell实验检测食管癌细胞增殖、迁移和侵袭,通过HE染色和免疫组化在移植瘤裸鼠体内验证TGF-β1/B7-H3轴。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析,两两比较采用LSD-t检验。结果与PBS处理比较,加入Treg细胞共培养促进食管癌细胞Eca-109、TE-1的增殖(24 h:0.93±0.18、0.99±0.18比0.63±0.12、0.63±0.14)、迁移[(74.67±4.16)、(61.00±2.65)比(55.67±5.13)、(37.33±2.08)个]、侵袭能力(105.70±2.13、63.67±3.51比77.33±3.22、55.33±2.52)和TGF-β1的表达(2.17±0.02、2.82±0.08比1.00±0.02、1.00±0.05);SB431542在体外有效抑制TGF-β1诱导的细胞增殖[(0.96±0.14)比(1.46±0.22)个]、迁移[(84.67±5.03)、(62.67±5.69)比(120.30±4.51)、(119.30±7.02)个]和侵袭能力(45.33±4.04、47.33±2.52比59.33±2.52、58.00±2.00);与正常人食管上皮细胞Het-1a比较,食管癌细胞Eca-109和TE-1中B7-H3 mRNA水平(2.65±0.04、3.30±0.27比1.00±0.03)增加(P均<0.05);与对照比较,TGF-β1可以进一步诱导食管癌细胞中B7-H3蛋白表达(1.57±0.10、1.41±0.02比1.00±0.02、1.00±0.06),而敲低B7-H3逆转TGF-β1诱导的致癌作用;此外,在裸鼠模型中也观察到相似的结果。结论Treg细胞通过上调TGF-β1和B7-H3表达促进食管癌细胞增殖、迁移和侵袭。Objective To investigate the effect and molecular mechanism of regulatory T(Treg)cells on the proliferation,migration and invasion of esophageal cancer cells.Methods Treg cells in the peripheral blood of patients were isolated by flow cytometry;transforming growth factorβ1(TGF-β1),TGF-β1/Smad inhibitor(SB431542),TGF-β1 overexpression(TGF-β1 OE)and B7 homolog 3(B7-H3)inhibition lentiviral vector(sh-B7-H3)were used to regulate the expression of TGF-β1 and B7-H3 in esophageal cancer cells;qRT-PCR and western blot were used to detect the expression levels of TGF-β1 and B7-H3;CCK-8 and Transwell were used to detect the proliferation,migration and invasion of esophageal cancer cells;the TGF-β1/B7-H3 axis was verified in xenograft nude mice by HE staining and immunohistochemistry.An independent sample t-test was used to compare two groups,a one-way analysis of variance was used to compare multiple groups and an LSD-t test was used for duo comparison between various groups.Results Compared with PBS,the co-culture of Treg cells promoted the proliferation(0.93±0.18,0.99±0.18 vs 0.63±0.12,0.63±0.14)and migration(74.67±4.16,61.00±2.65 vs 55.67±5.13,37.33±2.08),invasiveness(105.70±2.13,63.67±3.51 vs 77.33±3.22,55.33±2.52)and expression of TGF-β1(2.17±0.02,2.82±0.08 vs 1.00±0.02,1.00±0.05)of Eca-109 and TE-1 esophageal cancer cells;SB431542 effectively inhibited TGF-β1 induced cell proliferation(0.96±0.14 vs 1.46±0.22),migration(84.67±5.03,62.67±5.69 vs 120.30±4.51,119.30±7.02)and invasion(45.33±4.04,47.33±2.52 vs 59.33±2.52,58.00±2.00);Compared with Het-1a,B7-H3 mRNA levels were increased in Eca-109 and TE-1(2.65±0.04,3.30±0.27 vs 1.00±0.03).Compared with the control group,TGF-β1 further induced an increase in B7-H3 protein level in esophageal cancer cells(1.57±0.10,1.41±0.02 vs 1.00±0.02,1.00±0.06),and the knockdown of B7-H3 reversed the carcinogenic effect induced by TGF-β1.In addition,similar results were observed in a nude mouse model.Conclusion Treg cells promote esophageal ca

关 键 词：TREG细胞 TGF-Β1 B7-H3 食管癌 增殖 迁移和侵袭 

分 类 号：R735.1[医药卫生—肿瘤]