重楼皂苷Ⅰ调控分泌型磷蛋白1介导磷脂酰肌醇3-激酶/蛋白激酶B信号通路诱导肺腺癌凋亡抗肿瘤机制的研究  

A study on anti-tumor mechanism of secreted phosphoprotein 1-mediated phosphatidylinositol 3 kinase/protein kinase B signaling pathway induced apoptosis by polyphyllinⅠ

作　　者：牟海军 赵丹[2] 王显艳 姚淑娟[2] 石寒冰 郑红艳 李晶 MU Hai-jun;ZHAO Dan;WANG Xian-yan;YAO Shu-Juan;SHI Han-bing;ZHENG Hong-yan;LI Jing(Department of Respiratory and Critical Care Medicine,the Third Affiliated Hospital of Qiqihar Medical University,Qiqihar,Heilongjiang 161000,China;不详)

机构地区：[1]齐齐哈尔医学院附属第三医院呼吸与危重症医学科,黑龙江161000 [2]齐齐哈尔医学院医学技术学院

出　　处：《慢性病学杂志》2024年第11期1620-1624,共5页Chronic Pathematology Journal

基　　金：黑龙江省教育厅基金项目(2019-KYYWF-1254)

摘　　要：目的通过研究重楼皂苷Ⅰ(polyphyllinⅠ,PPⅠ)对人肺腺癌A549细胞凋亡、分泌型磷蛋白1(secreted phosphoprotein 1,SPP1)及磷脂酰肌醇3-激酶/蛋白激酶B(phosphatidylinositol 3-kinase/protein kinase B,PI3K/AKT)通路关键蛋白的影响,探讨PPⅠ调控SPP1基因介导PI3K/AKT信号通路诱导肺腺癌A549细胞凋亡抗肿瘤的作用机制。方法应用流式细胞及AO/EB双染细胞技术检测PPⅠ对肺腺癌A549细胞凋亡的影响。应用Western blot检测PPⅠ对肺腺癌A549细胞凋亡相关蛋白(Bax、Bcl-2)、SPP1及PI3K/AKT通路关键蛋白(PI3K、p-PI3K、p-Akt)表达的影响。结果(1)细胞凋亡结果:PPⅠ浓度为0.25 mg/L、0.50 mg/L、1.00 mg/L时肺腺癌A549细胞的凋亡率分别为(2.90±0.15)%、(14.25±1.56)%、(25.32±1.18)%,差异有统计学意义(P<0.05)。肺腺癌的A549细胞凋亡率与PPⅠ浓度呈正相关。(2)AO/EB双染结果:随着PPⅠ浓度的增加,正常细胞减少,凋亡细胞增多;细胞的体积变化趋势与药物浓度呈负相关。(3)Western blot实验结果:与空白对照组比较,PPⅠ组Bax表达增强(F=578.47,P<0.001),Bcl-2表达减弱(F=615.72,P<0.001),SPP1表达减弱(F=1099.40,P<0.001),p-PI3K表达下降(F=504.27,P<0.001)、p-Akt表达下降(F=446.84,P<0.001),并且变化幅度呈现剂量依赖效应,各组间比较,差异有统计学意义(P<0.05)。各组PI3K比较,差异无统计学意义(P>0.05)。结论PPⅠ可能通过下调SPP1表达,使PI3K/AKT信号通路的激活减弱,引起Bax表达增强,Bcl-2表达减弱,从而诱导细胞凋亡产生抗肿瘤作用。Objective To reveal the effect of polyphyllinⅠ(PPⅠ)on the apoptosis of lung adenocarcinoma A549 cells and the key proteins of secreted phosphoprotein 1(SPP1)and phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)pathway,and to explore the anti-tumor mechanism of PPⅠregulating SPP1 gene to induce apoptosis of lung adenocarcinoma A549 cells through PI3K/AKT signaling pathway.Methods Flow cytometry and AO/EB double staining were used to detect the effect of PPⅠon the apoptosis of lung adenocarcinoma A549 cells.Western blot was used to detect the effect of PPⅠon the expression of apoptosisrelated proteins(Bax,Bcl-2),SPP1 and PI3K/AKT pathway key proteins(PI3K,p-PI3K,p-Akt)in lung adenocarcinoma A549 cells.Results(1)The results of apoptosis:the apoptosis rates of lung adenocarcinoma A549 cells were(2.90±0.15)%,(14.25±1.56)%and(25.32±1.18)%,respectively,when the concentration of PPⅠwas 0.25 mg/L,0.50 mg/L and 1.00 mg/L,and the differences were statistically significant(P<0.05).And lung adenocarcinoma A549 cells apoptosis rate is positively related with PPⅠconcentration.(2)AO/EB results:with the increase of concentration of PPⅠ,normal cells decreased,apoptotic cells increased;The cell volume was negatively correlated with PPⅠconcentration.(3)Western blot:compared with control group,the expression of Bax in PPⅠgroup was increased(F=578.47,P<0.001),the expression of Bcl-2 was decreased(F=615.72,P<0.001),the expression of SPP1 was decreased(F=1099.40,P<0.001),the expression of p-PI3K(F=504.27,P<0.001)and p-Akt(F=446.84,P<0.001)decreased,the change showed a dose-dependent effect,the results were also statistically different between groups(P<0.05).There was no significant difference in PI3K among the groups(P>0.05).Conclusion PPⅠmay inhibit the activation of PI3K/AKT signaling pathway by down-regulating the expression of SPP1,increase the expression of Bax and decrease the expression of Bcl-2,thereby inducing cell apoptosis and exerting anti-tumor effect.

关 键 词：肺腺癌 重楼皂苷Ⅰ 细胞凋亡 分泌型磷蛋白1 磷脂酰肌醇3-激酶/蛋白激酶B信号通路 

分 类 号：R734.2[医药卫生—肿瘤]