丹参酚酸B靶向SOX2OT/miRNA-34a/SOX2轴抑制大肠癌细胞侵袭转移

Salvianolic acid B inhibits invasion and metastasis of colorectal cancer cells by targeting SOX2OT/miR-34a/SOX2 axis

目的:验证SOX2OT/miRNA(miR)-34a/SOX2信号轴对人大肠癌LOVO细胞侵袭转移的影响以及丹参酚酸B(SalB)通过该信号轴浓度依赖性抑制LOVO细胞侵袭转移。方法:双荧光素酶报告基因检测SOX2OT与miR-34a、miR-34a与SOX2的靶向结合;瞬时转染SOX2OT质粒和miR-34a mimic至LOVO细胞,RT-qPCR验证转染效率;RT-qPCR、Western blot检测SOX2OT、miR-34a与SOX2三者表达的相关性;划痕实验、Transwell实验检测SOX2OT/miR-34a/SOX2轴对LOVO细胞侵袭迁移的影响;MTT法检测丹参酚酸B对LOVO生长抑制作用;RT-qPCR检测低、中、高浓度丹参酚酸B处理下SOX2OT的表达;划痕实验、Transwell实验检测低、中、高剂量丹参酚酸B作用时细胞侵袭迁移情况。结果:LOVO细胞中SOX2OT与miR-34a、miR-34a与SOX2之间存在靶向结合;SOX2OT抑制miR-34a,上调SOX2;miR-34a可下调SOX2OT和SOX2表达;miR-34a逆转SOX2OT对细胞侵袭和迁移能力的增强作用;在一定浓度范围内丹参酚酸B量效相关性抑制LOVO细胞生长;丹参酚酸B浓度依赖性抑制LOVO细胞SOX2OT表达;丹参酚酸B抑制LOVO细胞侵袭转移,且随着浓度增加,抑制作用增强。结论:SOX2OT/miR-34a/SOX2信号轴与LOVO细胞侵袭转移密切相关,丹参酚酸B通过抑制该信号轴量效相关性抑制LOVO细胞侵袭转移。

Objective: To verify the effect of SOX2 OT/miR-34 a/SOX2 signal axis on the invasion and metastasis of human colorectal cancer LOVO cells, and that Salvianolic acid B(SalB) concentration-dependently inhibits the invasion and metastasis of LOVO cells through the signal axis. Methods: Dual Luciferase Reporter Gene Assay was used to verify the targeted binding of SOX2 OT to miR-34 a and miR-34 a to SOX2;SOX2 OT plasmid and miR-34 a mimic were transient transfected into LOVO cells, RT-qPCR was used to verify the transfection efficiency;RT-qPCR and Western blot were used to detect the correlation between SOX2 OT, miR-34 a and SOX2 expression;Scratch test and transwell test were used to detect the effects of SOX2 OT/miR-34 a/SOX2 axis on invasion and migration of LOVO cells;MTT assay was used to detect the inhibitory effect of Salvianolic acid B on LOVO growth;The expression of SOX2 OT was detected by RT-qPCR after treatment with low, medium and high dosages of Salvianolic acid B;Scratch test and transwell test were used to detect the invasion and migration of cells treated with Salvianolic acid B at low, medium and high dosages. Results: There were targeting relationship between SOX2 OT and miR-34 a, and between miR-34 a and SOX2 in LOVO cells;SOX2 OT inhibited miR-34 a and upregulated SOX2;MiR-34 a could down regulate the expression of SOX2 OT and SOX2;MiR-34 a reversed the enhancement of SOX2 OT on cell invasion and migration;Salvianolic acid B inhibited the growth of LOVO cells in dose-dependent manner within a certain concentration range;Salvianolic acid B inhibits SOX2 OT expression in a concentration dependent manner;Salvianolic acid B inhibited the invasion and metastasis of LOVO cells, and the inhibitory effect increased with the increase of concentration. Conclusion: SOX2 OT/miR-34 a/SOX2 signal axis is closely related to the invasion and metastasis of LOVO cells, Salvianolic acid B inhibits the invasion and metastasis of LOVO cells by inhibiting this signal axis.