盐酸小檗碱对宫颈癌HeLa细胞外泌体的影响及其机制  

Effect of berberine hydrochloride on exosomes of cervical cancer HeLa cells and its mechanism

作　　者：王景 葛静[1] 王旭[1] 房桂英[1] 代丽丽[1] 刘晶[1] WANG Jing;GE Jing;WANG Xu;FANG Guiying;DAI Lili;LIU Jing(Department of Gynecology and Obstetrics,First Hospital,Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第一医院妇产科,河北石家庄050030

出　　处：《吉林大学学报（医学版）》2020年第4期779-785,I0005,共8页Journal of Jilin University：Medicine Edition

基　　金：河北省卫健委医学科学研究项目资助课题(20190431)。

摘　　要：目的:观察盐酸小檗碱对宫颈癌HeLa细胞释放外泌体的干预作用,探讨其对癌细胞增殖和转移能力的影响。方法:将HeLa细胞分为对照组和低、中、高剂量盐酸小檗碱组,分别采用0、10、20和40 mg·L-1盐酸小檗碱干预HeLa细胞24 h,高速离心提取外泌体,Western blotting法检测外泌体标志物CD81和CD63及磷脂酰肌醇3-激酶(PI3K)/蛋白质丝氨酸苏氨酸激酶(Akt)信号通路相关蛋白表达水平,MTT比色法检测细胞体外生长抑制率,划痕实验检测细胞迁移率,Transwell小室实验检测穿膜细胞数。结果:与对照组比较,低、中和高剂量盐酸小檗碱组细胞外泌体中外泌体标志物CD81和CD63蛋白表达水平明显降低(P<0.05),细胞生长抑制率明显升高(P<0.05),细胞迁移率和穿膜细胞数明显降低(P<0.05),细胞中PI3K、Akt和磷酸化Akt(p-Akt)蛋白表达水平明显降低(P<0.05)。与低剂量盐酸小檗碱组比较,中和高剂量盐酸小檗碱组细胞外泌体中CD81和CD63蛋白表达水平明显降低(P<0.05),细胞生长抑制率明显升高(P<0.05),细胞迁移率和穿膜细胞数明显降低(P<0.05),细胞中PI3K、Akt和p-Akt蛋白表达水平明显降低(P<0.05)。与中剂量盐酸小檗碱组比较,高剂量盐酸小檗碱组细胞外泌体中CD81和CD63蛋白表达水平明显降低(P<0.05),细胞生长抑制率明显升高(P<0.05),细胞迁移率和穿膜细胞数明显降低(P<0.05),细胞中PI3K、Akt和p-Akt蛋白表达水平明显降低(P<0.05),具有浓度依赖性。结论:盐酸小檗碱能够抑制宫颈癌细胞外泌体分泌和癌细胞增殖,降低癌细胞的侵袭和转移能力,其作用机制可能与下调PI3K/Akt信号通路相关蛋白表达有关。Objective:To observe the intervention effect of berberine hydrochloride on the exosomes released by cervical cancer HeLa cells,and to explore its effects on the abilities of proliferation and metastasis of cancer cells.Methods:The cervical cancer HeLa cells were divided into control group and low,medium and high doses of berberine hydrochloride groups,and they were intervented with 0,10,20 and 40 mg·L-1 berberine hydrochloride for 24 h.High-speed centrifugation was used to extract the exosomes and Western blotting method was used to detect the expression levels of exosome markers CD81,CD63 and phosphatidylinositol 3-kinase(PI3K)/protein-serine-threonine kinase(Akt)signaling pathway related proteins.MTT assay was used to determine the inhibitory rates of growth of the HeLa cells.Scratch assay was used to measure the migration rate of cells.Transwell assay was used to detect the number of transmembrane cells.Results:Compared with control group,the expression levels of cervical cancer cell exosome markers CD81 and CD63 proteins in the exosomes in low,medium and high doses of berberine hydrochloride groups were significantly decreased(P<0.05),while the inhibitory rates of growth of cancer cells were increased significantly(P<0.05);the migration rates of cells and the number of transmembrane cells were decreased significantly(P<0.05),and the expression levels of PI3K,Akt and phosphorylated Akt(p-Akt)proteins in the cancer cells were significantly decreased(P<0.05).Compared with low dose of berberine hydrochloride group,the expression levels of CD81 and CD63 proteins in the exosomes in medium and high doses of berberine hydrochloride groups were significantly decreased(P<0.05),while the inhibitory rates of growth of cancer cells were increased significantly(P<0.05);the migration rates of cells and the number of transmembrane cells were decreased significantly(P<0.05),and the expression levels of PI3K,Akt and p-Akt proteins in the cancer cells were significantly decreased(P<0.05).Compared with medium dose of berberine

关 键 词：盐酸小檗碱 外泌体 宫颈肿瘤 磷脂酰肌醇3-激酶 蛋白质丝氨酸苏氨酸激酶 信号通路 

分 类 号：R737.33[医药卫生—肿瘤]