微小RNA Let-7/Lin28调节环与鼻咽癌血管新生的关系研究

Relationship between miRNA Let-7/Lin28 feedback loop and nasopharyngeal angiogenesis

摘要:

目的  探讨微小RNA Let-7/Lin28调节环与鼻咽癌促血管生成因子的关系，并研究这一关系的作用机制。方法  选取2012年4月-2014年在湖北省襄阳市中心医院耳鼻喉科就诊的鼻咽癌患者54例。另选取同期40例慢性鼻炎患者和32例健康者作为对照组。采用实时荧光定量PCR（RT-PCR）检测组织中Lin28A、Lin28B及Let-7a miRNA相对表达量，ELISA检测血清中血管内皮生长因子（VEGF）、碱性成纤维生长因子（bFGF）及内皮抑素（ES）水平，蛋白免疫印迹法（Western blot）检测H-Ras/PTEN/PI3K/Akt通路蛋白表达情况，分析miRNA Let-7a/Lin28调节环与血管新生因子的关系。结果  RT-PCR检测发现鼻咽癌患者Lin28A miRNA和Lin28B miRNA相对表达量均高于慢性鼻炎组和健康对照组（P <0.05），而Let-7a miRNA的相对表达量低于上述两组（P <0.05）；ELISA检测结果发现鼻咽癌患者组VEGF、bGFG、ES表达量均高于慢性鼻炎组和健康对照组（P <0.05）；Western blot检测发现鼻咽癌患者组病理组织中H-Ras、p-PI3K及p-Akt表达量高于慢性鼻炎患者组和健康对照组（P <0.05），而p-PTEN表达量低于上述两组（P <0.05）；相关性分析发现，Lin28A、Lin28B miRNA与VEGF、bFGF、H-Ras、p-PI3K及p-Akt呈正相关（P <0.05），Let-7a miRNA与VEGF、bFGF、H-Ras、p-PI3K及p-Akt呈负相关（P <0.05）。结论  miRNA Lin28/Let-7调节环与血管新生有定的关联性，而这一关联性可能与激活Ras/PI3K/PTEN/Akt通路有关。

Abstract:

Objective  To investigate the relationship between miRNA Let-7/Lin28 feedback loop and nasopharyngeal pro-angiogenesis factors, and to study the mechanism of its effect. Methods From April 2012 to 2014, 54 cases of nasopharyngeal carcinoma (NPC) therapied in ENT in Xiangyang Central Hospital were selected, and 40 cases of chronic rhinitis patients and 32 healthy subjects were selected as control groups. Expressions of Lin28A, Lin28B and Let-7a miRNA relative were detected by RT-PCR, and the levels of VEGF, bFGF and ES in serum were detected by ELISA. Expressions of protein in H-Ras/PTEN/PI3K/Akt pathway were detected by western blotting, and relationship between miRNA Let-7a/ Lin28 feedback loop and angiogenesis factors was analyzed. Results  Expression levels of Lin28A miRNA, Lin28B miRNA in NPC patients were relatively higher than the other two groups, and the relative expression of Let-7a miRNA was less than the other two groups (P < 0.05). ELISA test found that expressions of VEGF, bGFG, ES in NPC patients were higher than the control groups (P < 0.05). Expressions of H-Ras, p-PI3K and p-Akt in NPC patients pathology tissue were higher than the other two groups (P < 0.05), and expression of p-PTEN was less than the above two groups (P < 0.05). The correlation analysis showed that Lin28A, Lin28B miRNA were positively correlated with VEGF, bFGF, H-Ras, p-PI3K and p-Akt (P < 0.05), and Let-7a miRNA was negatively correlated with VEGF, bFGF, H-Ras, p-PI3K and p-Akt (P < 0.05). Conclusions  miRNA Lin28/Let-7 feedback loop has certain correlation with angiogenesis, and this association may related to the  activation of Ras/PI3K/PTEN/Akt pathway.