肾上腺髓质素对肾肿瘤细胞PI3K/Akt/eNOS/VEGF信号通路的作用

Effect of the adrenomedullin on the signaling pathway of PI3K/Akt/eNOS/VEGF in renal tumor cell

目的研究肾上腺髓质素(ADM)对肾肿瘤细胞磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(Akt)/内皮型一氧化氮合酶(e NOS)/血管内皮生长因子(VEGF)信号通路的影响。方法将肾肿瘤细胞分为对照组、ADM组、VEGF组,ADM+VEGF受体抑制剂组,ADM+ADM shRNA组,每组设置24 h、48 h、72 h 3个时间梯度,western-blot检测对照组和ADM组细胞总蛋白中PI3K、Akt、e NOS、VEGF表达;检测VEGF组,ADM+VEGF受体抑制剂组和ADM+ADM shRNA组细胞总蛋白中PI3K、Akt、e NOS的表达。结果 PI3K、Akt、e NOS、VEGF在ADM组各时间梯度组肾肿瘤细胞中表达明显高于其在对照组细胞的表达(P<0.05)。PI3K、Akt、e NOS在VEGF组和ADM+VEGF受体抑制剂组各时间组肾肿瘤细胞中表达明显高于其在对照组细胞中表达(P<0.05)。PI3K、Akt、e NOS在ADM+ADM shRNA组各时间组肾肿瘤细胞中表达与其在对照组细胞中表达差异无统计学意义(P>0.05)。结论 ADM可激活PI3K/Akt/e NOS/VEGF信号通路;VEGF受体抑制剂不能阻断ADM激活PI3K/Akt/e NOS信号通路。ADM一方面通过VEGF发挥肾肿瘤血管生成作用,另一方面通过其受体激活PI3K/Akt/e NOS信号通路参与肾肿瘤血管生成作用。ADM可能成为抗血管治疗肾肿瘤新的靶点。

Obj ective To investigate the effect of the adrenomedullin on the signaling pathway of PI3K/Akt/eNOS/VEGF in renal tumor cell.M ethods The kidney tumor cells were divided into five groups,control group,ADM group,VEGF group, ADM ＋VEGF receptor inhibitor group, ADM ＋ADM shRNA group, moreover, every group was redivided three time gradient groups （24 hours,48 hours and 72 hours）.The expression levels of PI3K,Akt,eNOS, VEGF in control group and ADM group and the expression levels of P3I K,Akt, eNOS in VEGF group,ADM＋VEGF receptor inhibitor group,ADM ＋ADM shRNA group were detected by Western Blot,respectively.Results The expressions levels of the PI3K,Akt,eNOS,VEGF in ADM groups were significantly higher than those of control group （ P〈0.05） .The expression levels of PI3K,Akt,eNOS in VEGF groups and ADM＋VEGF receptor inhibitor groups were significantly higher than those of control group （ P〈0.05）. However there were no significant differences in the expression levels of PI3K, Akt, eNOS between ADM ＋ADM shRNA groups and control group （ P〈0.05）.C onclusion ADM can activate PI3K/Akt/eNOS signaling pathway in renal tumor cell,which can not be blocked by VEGF receptor inhibitor.On one hand ADM regulates renal tumor angiogenesis by VEGF,on the other hand,by activating PI 3K/Akt/eNOS signaling pathway to participate in renal tumor angiogenesis.ADM may become a promising anti-angiogenic target in the treatment of renal tumor.