盐酸小檗碱对恶性黑素瘤细胞A375增殖、凋亡和迁移的影响  

Effects of berberine hydrochloride on the proliferation,apoptosis and migration of malignant melanoma A375 cells

作　　者：郑增光 王鹏雁 ZHENG Zeng-guang;WANG Peng-yan(Department of Pathology,Cancer Hospital of The University of Chinese Academy of Sciences(Zhejiang Cancer Hospital),Hangzhou 310022,China;Department of Critical Care Medicine,Zhejiang Hospital,Hangzhou 310013,China)

机构地区：[1]中国科学院大学附属肿瘤医院(浙江省肿瘤医院)病理科,杭州310022 [2]浙江医院重症医学科,杭州310013

出　　处：《浙江中西医结合杂志》2021年第1期18-21,共4页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省医药卫生科技项目(No.2018252439)。

摘　　要：目的探讨盐酸小檗碱对恶性黑素瘤细胞A375增殖、凋亡和迁移的影响。方法根据盐酸小檗碱浓度将人恶性黑素瘤A375细胞株分为四组:空白对照组(0μmol/L)、低剂量组(40μmol/L)、中剂量组(80μmol/L)和高剂量组(120μmol/L)。采用CCK8法检测各组A375细胞增殖情况,流式细胞仪检测细胞凋亡情况,通过Transwell法检测各组细胞的迁移能力,RT-PCR和Western blot检测A375细胞TLR4 mRNA及蛋白表达水平。结果盐酸小檗碱(0、40、80、120μmol/L)分别处理A375细胞后,随盐酸小檗碱浓度增加,A375细胞存活率逐渐降低[(100.00±4.76)%、(70.37±3.06)%、(48.33±0.64)%、(26.60±2.67)%,F=298.778,P<0.01];细胞凋亡率逐渐升高[(1.47±0.25)%、(2.56±0.41)%、(48.13±8.20)%、(67.99±7.99)%,F=101.764,P<0.01];细胞穿膜数逐渐降低[(331.67±7.64)个、(248.67±8.08)个、(155.33±5.03)个、(109.33±9.02)个,F=510.805,P<0.01];TLR4 mRNA/(β-actin)表达量逐渐降低[(1.00±0.02)、(0.85±0.01)、(0.73±0.01)、(0.63±0.01),F=428.200,P<0.01];TLR4蛋白/(β-actin)表达量逐渐降低[(1.00±0.02)、(0.80±0.01)、(0.63±0.01)、(0.21±0.01),F=2590.195,P<0.01]。结论盐酸小檗碱可以促进黑素瘤A375细胞凋亡,抑制A375细胞的增殖和迁移能力,存在剂量依赖关系,其抗肿瘤机制可能与抑制TLR4信号通路有关。Objective To explore the effects of berberine hydrochloride on the proliferation,apoptosis and migration of malignant melanoma A375 cells.Methods A375 cells were first treated with different concentrations of berberine hydrochloride(0,40,80,and 120μmol/L).Cell proliferation,apoptosis,and migration were then detected using the CCK8 assay,flow cytometry and Transwell assay,respectively.The mRNA and protein expression of tolllike receptor 4(TLR4)were determined by reverse transcription-polymerase chain reaction and Western blot.Results As the concentration of berberine hydrochloride increased(0,40,80,and 120μmol/L),the survival rate of A375 cells decreased[(100.00±4.76)%,(70.37±3.06)%,(48.33±0.64)%,(26.60±2.67)%,F=298.778,P<0.01],the apoptosis rate increased[(1.47±0.25)%,(2.56±0.41)%,(48.13±8.20)%,(67.99±7.99)%,F=101.764,P<0.01],the number of cells penetrating the membrane decreased[(331.67±7.64),(248.67±8.08),(155.33±5.03),(109.33±9.02),F=510.805,P<0.01],the relative expression of TLR4 mRNA/(β-actin)decreased[(1.00±0.02),(0.85±0.01),(0.73±0.01),(0.63±0.01),F=428.200,P<0.01]and the relative expression of TLR4 protein/(β-actin)decreased[(1.00±0.02),(0.80±0.01),(0.63±001),(0.21±0.01),F=2590.195,P<0.01].Conclusion Berberine hydrochloride promotes the apoptosis of melanoma A375 cells and inhibits the proliferation and migration of A375 cells in a dose-dependent manner.This effect may be related to the inhibition of the TLR4 signaling pathway.

关 键 词：盐酸小檗碱 黑色素瘤 A375细胞 增殖 凋亡 迁移 TLR4信号通路 

分 类 号：R285.5[医药卫生—中药学]