BBOX1-AS1被TFAP4激活从而促进胃癌细胞的增殖和迁移

BBOX1-AS1 is activated by TFAP4 to promote proliferation and migration of gastric cancer cells

目的:探讨BBOX1-AS1在胃癌中的分子功能和潜在作用机制。方法:实时定量PCR(qRT-PCR)检测BBOX1-AS1在胃癌细胞系中的表达水平。分别过表达和抑制TFAP4表达检测BBOX1-AS1表达水平,荧光素酶实验检测TFAP4和BBOX1-AS1启动子结合情况。CCK-8、EDU和划痕实验检测检测BBOX1-AS1对胃癌细胞系增殖、迁移的影响。Western blot检测迁移相关蛋白MMP-2、MMP-9的表达水平。结果:qRT-PCR结果显示,BBOX1-AS1在胃癌组织和细胞系中高表达(P<0.05),荧光素酶实验证实TFAP4通过与BBOX1-AS1启动子结合,转录调控BBOX1-AS1表达。CCK-8实验和EDU实验显示,抑制BBOX1-AS1表达可显著抑制MKN45细胞增殖(P<0.05)。划痕实验显示抑制BBOX1-AS1表达可降低MKN45细胞迁移能力(P<0.05)。Western blot实验结果表明,抑制BBOX1-AS1表达可显著降低MMP-2、MMP-9表达(P<0.05)。结论:BBOX1-AS1在胃癌组织和细胞系中高表达,被TFAP4转录激活,可促进胃癌细胞MKN45的增殖和迁移。

Objective:To investigate the function and potential mechanism of BBOX1-AS1 in gastric cancer.Methods:Expression of BBOX1-AS1 in gastric cancer cell line was detected by real-time quantitative PCR(qRT-PCR). Expression level of BBOX1-AS1 was detected by overexpression and inhibition of TFAP4,and the binding of TFAP4 and BBOX1-AS1 promoter were detected by luciferase assay. CCK-8,EDU and scratch test were used to detect effects of BBOX1-AS1 on proliferation and migration of gastric cancer cell lines. Western blot was used to detect expression levels of migration-related proteins MMP-2 and MMP-9.Results:qRT-PCR showed that BBOX1-AS1 was highly expressed in gastric cancer tissues and cell lines(P<0.05). Luciferase assay confirmed that TFAP4 transcriptionally regulated BBOX1-AS1 expression by binding to BBOX1-AS1 promoter. CCK-8 test and EDU test showed that inhibition of BBOX1-AS1 expression can significantly inhibited MKN45 cell proliferation(P<0.05). Scratch test showed that inhibition of BBOX1-AS1 expression can reduced migration ability of MKN45 cells(P<0.05). Western blot result showed that inhibition of BBOX1-AS1 expression can significantly reduce expressions of MMP-2 and MMP-9(P<0.05).Conclusion:BBOX1-AS1 is highly expressed in gastric cancer tissues and cell lines,and is transcriptionally activated by TFAP4,which can promote proliferation and migration of gastric cancer cell line MKN45.