白细胞介素-33增强人黑色素瘤A375细胞侵袭和增殖作用的机制研究  

Study on the Mechanism of Interleukin-33 Enhancing Invasion and Proliferation of Human Melanoma A375 Cells

作　　者：谢明水 李玲 邓涛 张秋莹 晏文强 黄宏耀 刘杨 XIE Ming-shui;LI Ling;DENG Tao;ZHANG Qiu-ying;YAN Wen-qiang;HUANG Hong-yao;LIU Yang(Department of Clinical Laboratory,Suizhou Hospital,Hubei University of Chinese Medicine,Suizhou 441300,China)

机构地区：[1]湖北医药学院附属随州医院,随州441300

出　　处：《微循环学杂志》2022年第2期1-7,共7页Chinese Journal of Microcirculation

基　　金：湖北省卫生计生科研项目(WJ2019M099)。

摘　　要：目的:研究白细胞介素-33(IL-33)对人黑色素瘤A375细胞侵袭和增殖的作用及机制。方法:采用ELISA试剂盒分别检测15例人黑色素瘤患者(患者组)和20例健康人群(健康对照组)血清中IL-33含量;将40只SPF级BALB/C裸鼠分为动物对照组、IL-33低、中、高剂量组,每组10只,构建人黑色素瘤A375细胞荷瘤鼠模型,分析IL-33在体内对肿瘤增长的影响,免疫组化分析肿瘤组织中血管内皮生长因子(VEGF)的表达水平。将对数生长期的人黑色素瘤A375细胞分为细胞对照组、1ng/ml IL-33组、3ng/ml IL-33组、30ng/ml IL-33组,通过CCK-8分析各组A375细胞增殖能力;Transwell和划痕实验分析A375细胞的侵袭能力和迁移能力;Western Blot分析细胞VEGF、波形蛋白(Vimentin)、黏钙蛋白N(N-cadherin)、黏钙蛋白E(E-cadherin)蛋白表达水平。结果:患者组血清IL-33水平明显高于健康对照组(P<0.01)。动物实验中,荷瘤鼠模型的肿瘤体积明显增大(P<0.05),免疫组化显示肿瘤组织中VEGF的表达水平也显著提高(P<0.01)。细胞实验中,与细胞对照组比较,不同剂量IL-33处理组人黑色素瘤A375细胞的增殖、侵袭和迁移能力显著增强,均呈现出显著的剂量依赖性(P<0.05),Vimentin、N-cadherin、VEGF表达明显增加,E-cadherin表达显著降低(P<0.01)。结论:IL-33能够通过调控人黑色素瘤A375细胞的上皮间质转化(EMT)机制和VEGF的表达,促进肿瘤细胞的增殖、侵袭和转移能力,促使肿瘤生长。Objective:To study the mechanism of interleukin-33(IL-33)enhancing invasion and proliferation of human melanoma A375 cells.Method:According to the retrospective analysis,the serum levels of IL-33 in 15 human melanoma patients(patient group)and 20 healthy people(healthy control group)were detected with ELISA kit.Human melanoma A375 cells at logarithmic growth stage were divided into control group,1ng/ml IL-33 group,3ng/ml IL-33 group and 30ng/ml IL-33 group to analyze the effects of different doses of IL-33 on the proliferation ability of human melanoma A375 cells by cell counting kit-8 in vitro.The influence of IL-33 on the invasion and migration ability of human melanoma A375 cells were analyzed by transwell chamber and scratch assay.The expression levels of VEGF,Vimentin,N-cadherin and E-cadherin proteins in A375 cells were analyzed by western blotting.Forty SPF BALB/C nude mice were divided into control group,low-dose,medium-dose and high-dose IL-33 groups,with 10 mice in each group constructed human melanoma A375 cells.The effect of IL-33 on tumor growth in vivo was analyzed,and the expression level of VEGF in tumor tissues was analyzed by immunohistochemistry.Results:The level of IL-33 in the serum of melanoma patient group was significantly higher than that in healthy control group(P<0.01).Compared with the control group,the proliferation,invasion and migration of melanoma A375 cells in different doses of IL-33 treatment group were significantly enhanced,all showed a significant dose-dependent(P<0.05).The expression of Vimentin,N-cadherin,VEGF increased significantly,and the expression of E-cadherin decreased significantly(P<0.01).The tumor volume of tumor-bearing mice increased significantly(P<0.05),immunohistochemistry showed that the expression level of VEGF in tumor tissues was also significantly increased(P<0.01).Conclusion:IL-33 can promote the proliferation,invasion and metastasis of tumor cells and promote tumor growth by enhancing the EMT mechanism and VEGF expression of human melanoma A375 cells.

关 键 词：白细胞介素-33 增殖 侵袭 转移 黑色素瘤 

分 类 号：R739.5[医药卫生—肿瘤]