基于PI3K/Akt信号通路探讨温下方乙酸乙酯部位对A549细胞移植瘤裸鼠肿瘤生长的抑制作用

Inhibitory effects of ethyl acetate fraction of Wenxia Formula on nude mice bearing transplanted A549 cells tumor via PI3K/Akt signaling pathway

目的探讨温下方乙酸乙酯部位经调控PI3K/Akt信号通路抑制A549细胞移植瘤生长的作用及机制。方法建立人肺腺癌A549细胞移植瘤裸鼠模型,随机分为模型组、阳性对照组(顺铂注射液,2.0 mg/kg)和温下方高、中、低剂量组(400、200、100 mg/kg),每组5只,药物干预5周后,取裸鼠肿瘤,称定质量并计算抑瘤率,HE染色观察裸鼠肿瘤病理形态学变化,TUNEL染色检测裸鼠肿瘤凋亡情况,Western blot法检测裸鼠肿瘤组织Akt、p-Akt、PI3K、p-PI3K、MMP-3、caspase-3、Bcl-2蛋白表达。结果与模型组比较,顺铂组和温下方各剂量组肿瘤质量和瘤组织p-Akt、p-PI3K、MMP-3、Bcl-2蛋白表达降低(P<0.05),凋亡阳性细胞面积比例和瘤组织caspase-3蛋白表达升高(P<0.05),肿瘤细胞出现不同程度的坏死。结论温下方乙酸乙酯部位可抑制裸鼠A549细胞移植瘤的生长,其机制可能与调控PI3K/Akt信号通路有关。

AIM To investigate the inhibitory effects and mechanism of ethyl acetate fraction of Wenxia Formula on the growth of transplanted A549 cells tumor via PI3K/Akt signaling pathway.METHODS The nude mouse models bearing transplanted human lung adenocarcinoma A549 cells tumor were randomly divided into the model group,the positive control group(cisplatin injection,2.0 mg/kg)and the high-dose,medium-dose and low-dose Wenxia Formula groups(400,200,100 mg/kg),with 5 mice in each group.After 5 weeks of drug intervention,the nude mice had their tumors excised and weighed,and the tumor inhibition rate calculated as well;their pathology of tumor observed by HE staining;their apoptosis of tumor detected by TUNEL staining;and their protein expressions of Akt,p-Akt,PI3K,p-PI3K,MMP-3,caspase-3 and Bcl-2 in tumor detected by Western blot.RESULTS Compared with the model group,the cisplatin group and the Wenxia Formula groups displayed decreased tumor weight and protein expressions of p-Akt,p-PI3K,MMP-3 and Bcl-2(P<0.05);increased area ratio of apoptotic positive cells and caspase-3 protein expression in the tumor tissue(P<0.05),and different degrees of tumor cells necrosis.CONCLUSION Wenxia Formula can inhibit the growth of transplanted A549 cells tumor in nude mice,and its mechanism may be related to its regulatory effect via PI3K/Akt signaling pathway.