文章摘要
王庆源,彭 稳,李 杰,顾奇偶,傅 赞.AGAP2⁃AS1调控Ras/MAPK信号通路促进结直肠癌细胞的增殖[J].南京医科大学学报,2019,(1):16~20
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
投稿时间:2018-10-26  
DOI:10.7655/NYDXBNS20190103
中文关键词: LncRNA  AGAP2⁃AS1  Ras/MAPK  增殖
英文关键词: LncRNA  AGAP2⁃AS1  Ras/MAPK  proliferation
基金项目:国家自然科学基金(81470881)
作者单位
王庆源 南京医科大学第一附属医院普外科江苏 南京 210029 
彭 稳 南京医科大学第一附属医院普外科江苏 南京 210029 
李 杰 南京医科大学第一附属医院普外科江苏 南京 210029 
顾奇偶 南京医科大学第一附属医院普外科江苏 南京 210029 
傅 赞 南京医科大学第一附属医院普外科江苏 南京 210029 
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中文摘要:
      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。
英文摘要:
      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.
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