紫草素对HaCat细胞增殖和AnxA2表达水平的影响  

Effects of Shikonin on the proliferation of HaCat cells and AnxA2 expression

作　　者：胡炜昊 阿依恒·曲库尔汗[1] HU Weihao;Ayiheng Qukuerhan(Department of Otorhinolaryngology,the First Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第一附属医院耳鼻喉科,乌鲁木齐830054

出　　处：《新疆医科大学学报》2021年第9期1048-1054,共7页Journal of Xinjiang Medical University

基　　金：新疆维吾尔自治区科技计划项目(2020E01055)。

摘　　要：目的探讨紫草素对HaCat细胞增殖和膜联蛋白A2(AnxA2)表达的影响。方法用不同浓度的紫草素(1、5、10μmol/mL)干预正常生长的HaCat细胞24 h,设0μmol/mL为正常对照组,镜下观察细胞生长状态;划痕试验检测HaCat细胞迁移;流式细胞术检测HaCat细胞的细胞周期;CCK-8细胞增殖检测实验(CCK-8法)检测HaCat细胞增殖;实时荧光定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹实验(Western blot)分别检测AnxA2 mRNA和蛋白的表达。结果与对照组相比,干预组紫草素浓度越高,细胞密度越低,细胞迁移速率越差;0、1、5、10μmol/mL组细胞G1期细胞百分比随紫草素浓度升高而升高,S期细胞百分比随紫草素浓度升高而降低,与对照组相比差异均有统计学意义(P<0.05);0、1、5、10μmol/mL组450 nm处吸光度值分别为1.28±0.15、1.15±0.12、0.94±0.08和0.65±0.01,与对照组相比差异有统计学意义(P<0.05);RT-qPCR示AnxA2表达水平随紫草素浓度升高呈剂量依赖性降低,与对照组相比差异有统计学意义(P<0.05);Western blot结果示AnxA2蛋白表达量随紫草素浓度增高而降低。结论紫草素可呈剂量依赖性地抑制HaCat细胞增殖并可降低AnxA2的表达水平,紫草素可能在治疗中耳胆脂瘤上皮细胞增殖过程中具有潜在的治疗作用。Objective To investigate the effects of shikonin on the proliferation of HaCat cells and annexin A2(AnxA2)expression in HaCat cells.Methods HaCat cells were incubated with different concentrations of shikonin(0,1,5,10μmol/mL).Growth status of HaCat cells was observed under microscope.Migration ability of HaCat cells was detected by scratch test.Cellular cycle of HaCat cells was measured through flow cytometry.Proliferation of HaCat cells was detected by Cell Counting Kit-8(CCK-8)method.AnxA2 mRNA and protein expression in all group were detected by RT-qPCR and Western blotting respectively.Results Compared with the control group,not only the cell density reduced but also cell migration ability became worse gradually with dose-dependence of shikonin.Also percentage of cells in G1 phase increased,and percentage of cells in S phase decreased with dose-dependence of shikonin(P<0.05).The absorbance value at 450nm of the 0,1,5,10μmol/mL groups was 1.28±0.15,1.15±0.12,0.94±0.08 and 0.65±0.01 respectively(P<0.05).RT-qPCR and Western blotting analysis showed that both AnxA2 mRNA and protein expression decreased in dose-dependence of shikonin(P<0.05).Conclusion Shikonin can inhibit proliferation of HaCat cells and reduce AnxA2 expression.This may have a potential therapeutic effect to the proliferation of epithelial cells in middle ear cholesteatoma.

关 键 词：紫草素 角质细胞 细胞增殖 膜联蛋白A2 

分 类 号：R764.29[医药卫生—耳鼻咽喉科]