【摘要】 目的 :探讨多发性内分泌腺瘤病1型(multiple endocrine neoplasia type 1,MEN 1)基因编码蛋白menin对人胃癌AGS细胞生长的影响,及其可能的作用机制。方法:将携带有MEN1基因的重组腺病毒Ad-MEN1感染人胃癌AGS细胞,然后采用蛋白质印迹法和免疫荧光法检测Ad-MEN1感染后对AGS细胞中menin表达水平的影响;分别采用MTT和FCM法检测menin过表达对AGS细胞增殖、细胞周期和凋亡的影响;再用蛋白质印迹法检测磷酸肌醇3-激酶(phosphoinositide 3-kinase,PI3K)/蛋白激酶B(protein kinase B,PKB,又称Akt)和核因子-κB(nuclear factor-kappa B,NF-κB)信号通路相关蛋白及其磷酸化水平的变化。结果:蛋白质印迹法和免疫荧光检测结果显示,menin蛋白在Ad-MEN1感染后的AGS细胞中表达水平明显增加(P<0.05),重组腺病毒Ad-MEN1在AGS细胞中的感染复数为60时,感染效率最佳,细胞增殖受到明显抑制(P<0.05)。DNA合成前期(G0/G1期)细胞所占比例上升,合成期(S期)细胞所占比例下降(P值均<0.05)。然而,menin高表达对AGS细胞的凋亡没有明显的作用(P>0.05)。Menin过表达后,AGS细胞中磷酸化Akt(phospho-Akt,p-Akt)和NF-κB p65的表达明显下调(P值均<0.05)。结论:Menin高表达可以抑制人胃癌AGS细胞的生长,其机制可能与抑制P13K/Akt和NF-κB信号转导通路有关。更多还原

【Abstract】 Objective: To investigate the effect of menin protein encoded by the multiple endocrine neoplasia type I(MEN1) gene on proliferation of human gastric cancer AGS cells and its possible machanism.Methods: An adenovirus encoding MEN1 gene was used to infect human gastric cancer AGS cells. The expression level of menin protein in AGS cells infected with recombinant adenovirus Ad-MEN1 was detected by Western blotting and immunof uorescence. The proliferation of AGS cells was tested by MTT method.The cell cycle distribution and apoptosis of AGS cells were analyzed by flow cytometry(FCM).The expressions of phospho-Akt(p-Akt) and nuclear factor-kappa B(NF- κB) which were related to phosphoinositide 3-kinase(PI3K)-protein kinase B(PKB)s ignal pathway were detected by Western blotting.Results: The results of Western blotting and immun of uorescence showed that the expression of menin protein in gastric cancer cell line AGS wasi ncreased after Ad-MEN1 infection( P <0.05). The infective effciency of recombinant adenovirus Ad-MEN1 was the highest when the multiplicity of infection was 60 in AGS cells, and the cell proliferation was obviously inhibited(P < 0.05). The percentage of cells was increased in phase G0/G1 and decreased in phase S(P < 0.05). However, the apoptosis rate of AGS cells after infection with Ad-MEN1 was not significantly changed(P > 0.05). The expressions of p-Akt and NF-κB p65 in AGS cells were decreased after the expression of menin in AGS cells was increased(P < 0.05).Conclusion: This study reveals that over-expression of menin can inhibit the growth of human gastric cancer cell line AGS. This machanism may be related to the inhibition of PI3K/Akt and NF-κB signal transduction pathways.更多还原