AGAP2-AS1调控Ras/MAPK信号通路促进结直肠癌细胞的增殖

AGAP2-AS1 promotes colorectal cancer proliferation through Ras/MAPK signaling pathway

目的:探讨长链非编码RNA(long noncoding RNA,LncRNA)AGAP2-AS1对结直肠癌细胞增殖能力的影响及其相关机制。方法:采用qRT-PCR检测50例结直肠癌组织和正常肠黏膜组织中AGAP2-AS1的表达水平;采用siRNA-AGAP2-AS1小干扰序列转染至人结直肠癌细胞系DLD-1和HT29细胞中,qRT-PCR检测细胞转染效率,CCK8实验和平板克隆形成实验检测AGAP2-AS1对细胞增殖能力的影响;流式细胞仪检测细胞周期分布的变化;Western blot检测MAPK通路相关蛋白(Ras、Raf-1、MEK、ERK)的表达。结果:与正常肠黏膜上皮组织相比,AGAP2-AS1的mRNA表达水平在结直肠癌中明显上调;下调AGAP2-AS1表达后,DLD-1和HT29细胞增殖能力降低,细胞周期阻滞于G0/G1期,Ras、p-Raf-1、p-MEK和p-ERK蛋白表达水平明显降低。结论:AGAP2-AS1通过调控Ras/MAPK通路促进结直肠癌细胞的增殖。

Objective:To explore the proliferation effect of long noncoding RNA(LncRNA)AGAP2-AS1 on colorectal cancer(CRC)cell lines and the associated signaling pathways. Methods:We conducted qRT-PCR to investigate mRNA levels of AGAP2-AS1 in 50 paired CRC tissues and the corresponding normal mucosa tissues. SiRNA targeting AGAP2-AS1 was transfected into DLD-1 and HT29 cell lines,qRT-PCR was performed to detect the transfection efficacy of siRNA and AGAP2-AS1 downregulated DLD-1 and HT29 cells were used for further investigating the role of AGAP2-AS1 in CRC cell proliferation through CCK8-kit and colony formation assay. Cell cycle distributions were analyzed by a flow cytometer. Western blot was conducted to explore the protein levels of MAPK signaling pathway,including Ras,Raf-1,MEK and ERK. Results:The mRNA levels of AGAP2-AS1 were markedly upregulated in CRC tissues rather than the corresponding normal tissues. Loss of AGAP2-AS1 greatly suppressed cell proliferation of DLD-1 and HT29 cells,increased percentage of G0/G1 phases population and downregulated the protein levels of Ras,p-Raf-1,p-MEK and p-ERK.Conclusion:LncRNA AGAP2-AS1 could promote colorectal cancer cell proliferation through Ras/MAPK signaling pathway.