白藜芦醇通过阻断JAK激酶2/信号转导及转录活化因子3信号通路抑制食管癌细胞增殖和迁移并诱导其凋亡的作用研究  

Effect of resveratrol on inhibiting proliferation,migration and inducing apoptosis of esophageal cancer cells by blocking JAK kinase 2/signal transduction and transcription activation factor 3 signaling pathway

作　　者：崔晓佳[1] 谭程[1] 杨百霞[1] 倪峰[1] 杭达明[1] 沈健 钱霞[1] CUI Xiaojia;TAN Cheng;YANG Baixia;NI Feng;HANG Daming;SHEN Jian;QIAN Xia(Department of Radiotherapy,Nantong Tumor Hospital,Nantong,Jiangsu 226361,China)

机构地区：[1]南通市肿瘤医院放疗科,江苏南通226361

出　　处：《安徽医药》2024年第6期1103-1108,共6页Anhui Medical and Pharmaceutical Journal

基　　金：2018年度南通市市级科技计划(指导性)项目(MSZ18127);2022年南通市卫生健康委员会科研课题(MSZ2022026)。

摘　　要：目的探究白藜芦醇对人食管癌细胞增殖、凋亡、迁移及JAK激酶2/信号转导及转录活化因子3(JAK2/STAT3)信号通路的调控作用。方法2021年7月至2022年7月,体外培养人食管癌OE19细胞,将细胞分为对照组(不做干预)和实验组(15、30、60、90和120μmol/L白藜芦醇干预24 h)进行预实验,根据细胞计数试剂盒(CCK-8)预实验结果,筛选有显著作用且细胞活力高于50%的30、60、90μmol/L白藜芦醇进行后续的实验,后续实验又分为对照组(不做干预)、30、60、90μmol/L白藜芦醇组(30、60和90μmol/L白藜芦醇)和30μmol/L白藜芦醇+抑制剂组(30μmol/L白藜芦醇+10μmol/L JAK2/STAT3通路抑制剂AG490),干预24 h。用5-乙炔基-2'脱氧尿嘧啶核苷(EdU)、Hoechst 33258染色、Transwell、实时荧光定量PCR(RT-qPCR)及蛋白质印迹法对细胞增殖率、凋亡情况、迁移数及细胞周期蛋白D1(cyclin D1)、胱天蛋白酶-3(caspase-3)和JAK2/STAT3相关因子表达水平进行分析。结果不同浓度实验组的细胞活力与对照组相比逐渐降低,其中30、60、90和120μmol/L白藜芦醇差异有统计学意义(P<0.05),但120μmol/L白藜芦醇组细胞活力低于50%,所以本研究选择30、60和90μmol/L白藜芦醇继续后续实验;60μmol/L白藜芦醇组细胞增殖率(41.49±5.06)%、迁移细胞数(36.67±2.52)个显著低于对照组(53.34±1.99)%、(58.00±2.00)个,凋亡细胞数(7.67±1.53)个显著高于对照组(2.50±0.71)个,且cyclin D1、p-JAK2和p-STAT3表达量也低于对照组,caspase-3 mRNA和蛋白表达水平高于对照组(P<0.05);30、90μmol/L白藜芦醇组以上各指标与对照组比较同样如此。与30μmol/L白藜芦醇组相比,30μmol/L白藜芦醇+抑制剂组以上各指标变化更显著(P<0.05)。结论白藜芦醇可通过抑制JAK2/STAT3通路抑制人食管癌OE19细胞的增殖和迁移并诱导凋亡。Objective To explore the regulatory effects of resveratrol on the proliferation,apoptosis,migration and JAK kinase 2/signal transduction and transcriptional activation factor 3(JAK2/STAT3)signaling pathway of human esophageal cancer cells.Methods From July 2021 to July 2022,human esophageal cancer OE19 cells were cultured in vitro,and the cells were divided into control group(without intervention)and experimental group(15,30,60,90 and 120μmol/L resveratrol intervention for 24 h)for pre-experiment.According to the pre-experiment results of cell counting kit 8(CCK-8),30,60,90μmol/L resveratrol with significant effect and cell viability higher than 50%were screened for subsequent experiments.Follow-up experiments were divided into control group(no intervention),30,60,90μmol/L resveratrol group(30,60 and 90μmol/L resveratrol)and 30μmol/L resveratrol+inhibitor group(30μmol/L resveratrol+10μmol/L)JAK2/STAT3 pathway inhibitor AG490)was treated for 24 h.Cell proliferation rate,apoptosis,migration and cyclin D1 caspase-3 and JAK2/STAT3-related factors expression were measured by 5-acetylidene-2'deoxyuracil(EdU),Hoechst 33258 staining,Transwell,RT-qPCR and Western blotting.Results Compared with the control group,the cell viability of the experimental group with different concentrations decreased gradually,and the difference of 30,60,90 and 120μmol/L resveratrol was statistically significant(P<0.05),but the cell viability of 120μmol/L resveratrol group was lower than 50%.Therefore,30,60 and 90μmol/L resveratrol was selected for subsequent experiment in this study.The cell proliferation rate(41.49±5.06)%and the number of migrating cells(36.67±2.52)in 60μmol/L resveratrol group were significantly lower than those in control group(53.34±1.99)%and(58.00±2.00).The number of apoptotic cells(7.67±1.53)was significantly higher than that in control group(2.50±0.71),and the expressions of cyclin D1,p-JAK2 and p-STAT3 in 60μmol/L resveratrol group were lower than those in control group,and the mRNA and protein expres

关 键 词：食管肿瘤 白藜芦醇 JAK激酶2/信号转导及转录活化因子3信号通路 增殖 凋亡 迁移 

分 类 号：R285[医药卫生—中药学]