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

doi:10.3969/j.issn.2095-4344.0529

Abstract

Abstract:

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 siRNA 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 mRNA 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 (P < 0.05). The clone formation rate of p75^NTR positive cells was significantly higher than that of p75^NTR negative cells (P < 0.0.5). After 72 hours of transfection, the expression of SATB1 in the transfection group was significantly lower than that in the control group (P < 0.05). The ability of migration and invasion in the transfection group was significantly lower than that in the control group (P < 0.05). Expressions of matrix metalloproteinase 2/9 mRNA and protein in the transfection group were significantly lower than those of the control group (P < 0.05). In conclusion, siRNA interference with SATB1 gene can reduce the invasion and migration ability of TE-1 tumor stem cells in esophageal carcinoma cell line through down-regulating the expression of matrix metalloproteinase 2/9.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Esophageal Neoplasms, Neoplastic Stem Cells, RNA Interference, Lymphocytes, Tumor-Infiltrating, Tissue Engineering

CLC Number:

R394.2

Li Xiao-lei, Xu Jun, Hu Xin, He Wen-wu, Zheng Yin-bin. Silencing of SATB1 inhibits the invasion and migration of tumor stem cells in TE-1 cell line of esophageal squamous cell carcinoma[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(17): 2674-2679.

Figures/Tables 1

References

[1] Sun H, Zou S, Zhang S, et al. Elevated DNA polymerase iota (Poli) is involved in the acquisition of aggressive phenotypes of human esophageal squamous cell cancer. Int J Clin Exp Pathol.2015;8(4):3591-601.

[2] Chen P, Fang M, Wan Q, et al. High-sensitivity modified Glasgow prognostic score (HS-mGPS) Is superior to the mGPS in esophageal cancer patients treated with chemoradiotherapy. Oncotarget.2017;8(59):99861-99870.

[3] 刘盈君,刘守钦,王家林,等.济南市2011-2015年食管癌流行特征分析[J].中华肿瘤防治杂志,2017,24(3):147-150.

[4] 艾军华,时军.肿瘤干细胞和上皮-间质转化之间的关系及其在肿瘤侵袭转移中的作用[J].医学综述,2015,21(12):2180-2183.

[5] 李远强,张坤,孙丽丹.肿瘤干细胞靶向给药系统的研究进展[J].海南医学,2017,28(2):260-262.

[6] Tabe Y, Konopleva M. Leukemia stem cells microenvironment. Adv Exp Med Biol. 2017;1041:19-32.

[7] Wang T, Fahrmann JF, Lee H, et al.JAK/STAT3-regulated fatty acid β-Oxidation is critical for breast cancer stem cell self-renewal and chemoresistance. Cell Metab. 2018;27(1): 136-150.

[8] 李安.艾迪注射液联合FOLFOX4化疗对晚期结肠癌患者肿瘤干细胞特性及抗肿瘤免疫应答的影响[J].海南医学院学报,2017, 23(8):1113-1116.

[9] 廖星芸,汪洋,孙建国,等.IR-780联合阿霉素对肝癌干细胞样细胞的杀伤效应研究[J].第三军医大学学报,2017,39(12): 1206-1212.

[10] 朱海振,耿涛,李雪涛,等.利用分子信标检测microRNA并成像肺癌干细胞的实验研究[J].重庆医学,2016,45(15):2036-2039.

[11] 佟冬冬,张风河,姚瑶,等.p75神经营养蛋白受体阳性舌鳞状细胞癌细胞的生物学特性研究[J].华西口腔医学杂志,2014,(1): 18-22.

[12] 佟冬冬.p75神经营养因子受体作为舌鳞癌干细胞表面标志物的实验研究[J].山东大学,2012.

[13] 王峰.p75<'NTR>作为食管鳞癌干细胞膜蛋白标志的实验研究[D].第二军医大学,2009.

[14] 孙志刚,黄盛东,张宝仁,等.应用p75NTR分选食管肿瘤干细胞并鉴定其生物学特性[J].第二军医大学学报,2009,30(5):481-486.

[15] Wang Q, Hu SC, Yang CS, et al. Inhibition of prostate cancer cell growth in vivo with short hairpin RNA targeting SATB1. Oncol Lett.2017;14(6):6592-6596.

[16] Tanaka Y, Sotome T, Inoue A, et al.SATB1 conditional knockout results in sjögren's syndrome in mice.J Immunol. 2017;199(12):4016-4022.

[17] Choudhary D, Clement JM, Choudhary S, et al.SATB1 and bladder cancer: Is there a functional link? Urol Oncol. 2018; 36(3):93.e13-93.e21

[18] Lee JJ, Kim M, Kim HP. Epigenetic regulation of long noncoding RNA UCA1 by SATB1 in breast cancer.BMB Rep. 2016;49(10):578-583.

[19] 黄盛东,刘晓红,袁扬,等.食管癌中p75NTR阳性细胞池的变化与顺铂耐药机制的研究[J].中华实验外科杂志,2007,24(12): 1560-1562.

[20] 瞿丛新,吴莹莹.SATB1和E-cad在乳腺癌中的表达及其与淋巴结转移的关系[J].检验医学与临床,2017,14(8):1116-1118.

[21] 吴冬梅.SATB1表达调控肝癌转移能力的临床病理学意义[D].南昌大学,2015.

[22] 高超,朱艳卿,刘颖,等.SATB1基因在结直肠癌中的表达及其临床意义[J].肿瘤防治研究,2013,40(2):159-163.

[23] 吴晓飞,朱朝辉,万锋,等.AT富集序列特异性结合蛋白1在人膀胱移行细胞癌组织中的表达及其与上皮间质转化的关系[J].中华实验外科杂志,2013,30(6):1145-1148.

[24] 王坤,颜克强,刘承,等.SATB1在肾透明细胞癌中的表达及临床意义[J].中国现代医学杂志,2011,21(25):3106-3109,3115.

[25] 刘康,李杰,王兰,等.SATB1基因在食管癌中的表达及临床意义[J].川北医学院学报, 2015,30(4):447-450.

[26] Shi ZM, Liu YN, Fu B, et al. Expression profile of eukaryotic translation initiation factor and matrix metalloproteinase 9 in endometrial cancer tissue. J Biol Regul Homeost Agents. 2017;31(4):1053-1059.

[27] Weng H, Zeng XT, Wang XH, et al. Genetic association between matrix metalloproteinases gene polymorphisms and risk of prostate cancer: a meta-analysis. Front Physiol.2017; 8:975.