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目的研究丝氨酸/苏氨酸蛋白激酶(Akt)在胃癌中的表达、活化情况及其与血管内皮生长因子C(VEGFC)的关系,以初探Akt表达和活化的生物学意义及其可能的机制。方法采用RTPCR法检测20例新鲜胃癌及癌旁正常组织标本中Akt1、Akt2和Akt3mRNA的表达;Western印迹法检测Akt和磷酸化Akt(pAkt)蛋白的表达;免疫组化检测pAkt蛋白和VEGFC在55例胃癌配对组织中的原位表达。结果20例胃癌及相应癌旁正常组织中Akt1、2、3mRNA的表达阳性率皆为100%,且三者在癌和正常组织中的表达水平差异无统计学意义(P>0.05)。Western印迹显示Akt蛋白在胃癌组织中的表达水平为正常组织的2.7倍(P<0.001),而pAkt蛋白的表达水平为癌旁正常组织的4.1倍(P<0.01)。免疫组化分析表明,pAkt表达于67.3%(37/55)的胃癌组织,且较高表达于组织分化较差(χ2=9.751,P<0.01)和TNM分期较晚(χ2=7.684,P<0.05)者;VEGFC在胃癌组织中的表达阳性率为61.8%(34/55),其表达与肿瘤的TNM分期(χ2=9.298,P<0.01)及淋巴结转移(χ2=7.605,P<0.05)密切相关,统计分析表明,pAkt与VEGFC的表达呈显著性正相关(Pearsonr=0.524,P<0.05,Spearman,rho=0.747,P<0.01)。结论Akt及pAkt蛋白特异性过表达于胃癌组织,可能是蛋白翻译水平和(或)代谢水平改变的结果。Akt蛋白的磷酸化可能促进胃癌细胞的恶性转化和淋巴结转移,VEGFC可能在其中发挥重要的介导作用。
Objective To investigate the expression and activation of serine / threonine protein kinase (Akt) in gastric carcinoma and its relationship with vascular endothelial growth factor C (VEGFC) in order to explore the biological significance of Akt expression and activation and its possible mechanism. Methods RTPCR was used to detect the expression of Akt1, Akt2 and Akt3 mRNA in 20 cases of fresh gastric cancer tissues and adjacent normal tissues. The expressions of Akt and phosphorylated Akt (pAkt) proteins were detected by Western blotting. The expressions of pAkt protein and VEGFC were detected by immunohistochemistry Example gastric cancer paired tissue in situ expression. Results The positive rates of Akt1,2,3 mRNA expression in all 20 gastric cancer tissues and adjacent normal tissues were 100%. There was no significant difference in the expression of Akt1,2,3 mRNA between normal and cancerous tissues (P> 0.05). Western blotting showed that Akt protein expression in gastric cancer tissue was 2.7-fold higher than that in normal tissue (p <0.001), while that of pAkt protein was 4.1-fold (P <0.01). Immunohistochemical analysis showed that pAkt was expressed in 67.3% (37/55) of gastric cancer tissues, with higher expression in poorly differentiated tissues (χ2 = 9.751, P <0.01) and later stage of TNM (χ2 = 7.684, P < 0.05). The positive expression rate of VEGFC in gastric cancer tissues was 61.8% (34/55), which was significantly correlated with TNM stage (χ2 = 9.298, P <0.01) and lymph node metastasis (χ2 = 7.605, Statistical analysis showed that there was a significant positive correlation between pAkt and VEGFC (Pearson r = 0.524, P <0.05, Spearman, rho = 0.747, P <0.01). Conclusion The overexpression of Akt and pAkt protein in gastric cancer may be the result of changes in protein translation and / or metabolism. Phosphorylation of Akt protein may promote malignant transformation of gastric cancer cells and lymph node metastasis, VEGFC may play an important mediating role.