Silencing of SATB1 inhibits the invasion and migration of tumor stem cells in TE-1 cell line of esophageal squamous cell carcinoma / 中国组织工程研究

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: p75NTRpositive cells and p75NTRnegative cells were isolated from human esophageal carcinoma TE-1 cell lines by immunomagnetic beads. The characteristics of p75NTRpositive cells were verified by in vitro proliferation and clone formation experiments. The p75NTRpositive 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 p75NTRpositive 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 p75NTRnegative cells, the proliferation of p75NTRpositive cells increased significantly after 3, 5, 7 days of culture (P < 0.05). The clone formation rate of p75NTRpositive cells was significantly higher than that of p75NTRnegative 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.