蛋白激酶B磷酸化水平在非小细胞肺癌吉非替尼耐药中的临床意义

Clinical significance of protein kinase B phosphorylation levels in gefitinib resistance for non-small cell lung cancer

目的探讨蛋白激酶B(AKT)磷酸化及其信号通路在非小细胞肺癌(NSCLC)对吉非替尼耐药过程中的作用。方法用免疫组化方法检测肺癌组织芯片阵列组织中AKT和磷酸化AKT(p-AKT)的水平,并分析其与肺癌临床生物学特征、表皮生长因子受体(EGFR)、患者预后的关系。将细胞分为A549细胞组、A549-Ge耐药细胞组和A549-Ge耐药细胞+Perifosine组(0.5μmol·L^(-1)Perifosine)。用蛋白质印迹法检测细胞的AKT和p-AKT水平,研究p-AKT磷酸化水平对吉非替尼杀伤活性的影响。结果92例NSCLC标本,免疫组化检测发现肺癌组织AKT和p-AKT阳性水平均高于癌旁肺组织(AKT:61.96%vs 34.78%,P<0.05;p-AKT:53.26%vs 31.52%,P<0.05),且肺癌组织p-AKT的表达与临床分级、EGFR、间变性淋巴瘤激酶(ALK)、患者总生存期密切相关(均P<0.05),在临床分级Ⅱ~Ⅲ级、EGFR阳性、ALK阳性的患者中阳性率更高,高表达p-AKT的患者总生存期明显缩短。A549细胞组、A549-Ge耐药细胞组、A549-Ge耐药细胞+Perifosine组的p-AKT表达水平分别为(62.41±5.48)%、(126.85±8.34)%和(18.54±2.68)%,AKT表达水平分别为(29.56±4.52)%、(103.29±7.92)%和(23.16±2.64)%;A549-Ge耐药细胞组的上述指标与A549细胞组比较,A549-Ge耐药细胞+Perifosine组的上述指标与A549-Ge耐药细胞组比较,差异均有统计学意义(均P<0.05)。结论NSCLC细胞内AKT信号通路在决定肿瘤细胞是否对EGFR靶向药敏感中发挥重要作用。

Objective To explore the role of protein kinase B(AKT)phosphorylation and its signaling pathway in gefitinib resistance in non-small cell lung cancer(NSCLC).Methods Immunohistochemical method were used to detect the levels of AKT and phosphorylated AKT(p-AKT)in chip array tissues of lung cancer,and to analyze the relationship between AKT and clinical biological characteristics,epidermal growth factor receptor(EGFR),and prognosis of patients.The cells were divided into A549 group,A549-GE resistant group and A549-GE resistant+Perifosine group(0.5μmol·L^(-1) Perifosine).The levels of AKT and p-AKT were detected by western blotting,the effect of AKT phosphorylation on the killing activity of gefitinib was investigated.Results Immunohistochemical analysis of 92 NSCLC samples showed that the positive levels of AKT and p-AKT in lung cancer tissues were higher than those in adjacent lung tissues(AKT:61.96%vs 34.78%,P<0.05;p-AKT:53.26%vs 31.52%,P<0.05).The expression of p-AKT in lung cancer tissues was closely correlated with clinical grade,EGFR,anaplastic lymphoma kinase(ALK),and overall survival(all P<0.05).The positive rate was higher in patients with clinical gradeⅡ-Ⅲ,EGFR positive and ALK positive,and overall survival was significantly shortened in patients with high expression of p-AKT.The levels of p-AKT in A549 group,A549-GE resistant group and A549-GE resistant+Perifosine group were(62.41±5.48)%,(126.85±8.34)%and(18.54±2.68)%,respectively;the expression levels of AKT were(29.56±4.52)%,(103.29±7.92)%and(23.16±2.64)%,respectively.There were statistically significant differences between A549-GE resistant group and A549 group,and between A549-GE resistant+Perifosine group and A549-GE resistant group(all P<0.05).Conclusion The AKT signaling pathway in NSCLC cells plays an important role in determining whether tumor cells are sensitive to EGFR-targeting drugs.