SATB1基因沉默抑制食管鳞状细胞癌TE-1中肿瘤干细胞样细胞侵袭及迁移能力  

Silencing of SATB1 inhibits the invasion and migration of tumor stem cells in TE-1 cell line of esophageal squamous cell carcinoma

作　　者：李晓雷 徐俊 胡欣 何文武 郑银彬 Li Xiao-lei;Xu Jun;Hu Xin;He Wen-wu;Zheng Yin-bin(Department of Thoracic Surgery,2Department of Oncology,Nanchong Central Hospital,Nanchong 637000,Sichuan Province,China)

机构地区：[1]南充市中心医院.川北医学院第二临床医学院胸外科,四川省南充市637000 [2]南充市中心医院.川北医学院第二临床医学院肿瘤科,四川省南充市637000

出　　处：《中国组织工程研究》2018年第17期2674-2679,共6页Chinese Journal of Tissue Engineering Research

基　　金：四川省卫生和计划生育委员会科研课题(16PJ210);课题名称:SATB1靶向调控FN1基因对食管癌TE-1细胞侵袭迁移的影响及其分子机制研究~~

摘　　要：背景:近来有研究证实SATB1与肿瘤的发生及发展有关,但其在肿瘤转移中的机制尚不明确。目的:观察特异性干扰SATB1基因表达对食管鳞状细胞癌TE-1中肿瘤干细胞样细胞侵袭、迁移的影响。方法:采用免疫磁珠分选法从人食管鳞状细胞癌TE-1中分选p75^(NTR)阳性细胞与p75^(NTR)阴性细胞,检测p75^(NTR)阳性细胞的体外增殖能力及克隆形成能力。取对数生长期的p75^(NTR)阳性肿瘤干细胞样细胞,转染组利用Lipofectamine 2000脂质体法将SATB1基因si RNA转染至细胞中,同时设置空载体转染的p75^(NTR)阳性细胞为对照,转染后72 h,采用Western blot法检测2组细胞SATB1蛋白表达,Transwell小室实验检测2组细胞的迁移及侵袭能力,Western blot与q RT-PCR法检测基质金属蛋白酶2/9的表达。结果与结论:(1)与p75^(NTR)阴性细胞比较,p75^(NTR)阳性细胞培养3,5,7 d的细胞增殖明显加快(P<0.05);(2)p75^(NTR)阳性细胞的克隆形成率明显高于p75^(NTR)阴性细胞(P<0.05);(3)转染后72 h后,转染组SATB1基因蛋白表达明显低于对照组(P<0.05),迁移能力与侵袭能力明显低于对照组(P<0.05),基质金属蛋白酶2/9的基因及蛋白表达明显低于对照组(P<0.05);(4)结果提示,以si RNA干扰SATB1基因后,可抑制食管鳞状细胞癌TE-1肿瘤干细胞样细胞侵袭、迁移,可能与其下调基质金属蛋白酶2/9的表达有关。BACKGROUND：Recent studies have confirmed that SATB1 is related to the occurrence and development of tumors,but its mechanism in tumor metastasis is unclear.OBJECTIVE：To observe the effects of specific interference of SATB1 gene expression on esophageal carcinoma cell line TE-1 stem cell invasion and migration.METHODS：p75^NTR positive cells and p75^NTR negative cells were isolated from human esophageal carcinoma TE-1 cell lines by immunomagnetic beads.The characteristics of p75^NTR positive cells were verified by in vitro proliferation and clone formation experiments.The p75^NTR positive tumor stem cells in logarithmic growth period were taken.In the transfection group,SATB1 gene si RNA was transfected into the cells by Lipofectamine 2000 liposome method.At the same time,the p75^NTR positive cells transfected with empty vector were used as control.After 72 hours of transfection,the expression of SATB1 in the cells was detected by western blot.Cell migration and invasion abilities were detected by Transwell assay.Expressions of matrix metalloproteinase 2/9 at m RNA and protein levels were detected by western blot and semi-quantitative RT-PCR,respectively.RESULTS AND CONCLUSION：Compared with p75^NTR negative cells,the proliferation of p75^NTR positive cells increased significantly after 3,5,7 days of culture（P0.05）.The clone formation rate of p75^NTR positive cells was significantly higher than that of p75^NTR negative cells（P0.0.5）.After 72 hours of transfection,the expression of SATB1 in the transfection group was significantly lower than that in the control group（P0.05）.The ability of migration and invasion in the transfection group was significantly lower than that in the control group（P0.05）.Expressions of matrix metalloproteinase 2/9 m RNA and protein in the transfection group were significantly lower than those of the control group（P0.05）.In conclusion,si RNA interference with SATB1 gene can reduce the invasion and migration ability of TE-1 tumor stem cells in esophageal carcinoma

关 键 词：食管肿瘤 肿瘤干细胞 RNA干扰 淋巴细胞 肿瘤浸润 组织工程 食管鳞状癌 SATB1基因 细胞侵袭 细胞迁移 基质金属蛋白酶 基因沉默 Transwell小室 细胞转染 克隆形成 p75^(NTR) 

分 类 号：R394.2[医药卫生—医学遗传学]