摘要
目的探讨IL-33通过对NF-κB信号通路及炎症因子的调控,对肝癌HepG2细胞增殖和迁移的影响。方法RT-PCR检测IL-33在不同肝癌细胞系中的表达,Western Blot检测IL-33对HepG2细胞NF-κB信号通路激活相关蛋白(NF-κB、p-NF-κB、IKB、p-IKB)的调控,ELISA检测IL-33对HepG2细胞中IL-1α、IL-1β、IL-6、IL-18等炎症因子释放的调控,EdU增殖实验检测IL-33对HepG2细胞增殖的影响,CCK8实验检测IL-33对HepG2细胞24 h和48 h活力的影响,Transwell小室实验和划痕实验检测IL-33对HepG2细胞迁移的影响。结果IL-33 mRNA表达分别为LO2(1.01±0.01)、MHCC97H(1.08±0.05)、LM3(1.76±0.21)、HepG2(3.88±0.35)、SMCC7721(2.24±0.43)和Hep3B(2.13±0.30),差异有统计学意义(F=19.621,P=0.001)。IL-33处理24 h组中IL-1α(3185.25±194.67)、IL-1β(2103.69±124.75)、IL-6(446.58±36.82)、IL-18(1492.60±180.31),高于对照组的1001.58±18.65、1642.81±45.66、151.34±20.13、915.48±36.15,差异有统计学意义(P<0.05)。IL-33处理24 h组中NF-κB磷酸化蛋白(4.53±0.36)及IKB磷酸化蛋白(2.76±0.28)相对表达水平均高于对照组(1.25±0.23、0.49±0.09),差异有统计学意义(P<0.05)。IL-33处理24 h组细胞增殖(1.41±0.08)、24 h细胞活力(135.69±6.88)%、48 h细胞活力(176.34±25.69)%,高于对照组的(0.86±0.04)、(98.65±1.05)%、(119.89±10.86)%,差异有统计学意义(t=13.750,P<0.01;t=9.218,P<0.01;t=3.506,P=0.017)。Transwell小室和划痕实验结果发现,IL-33处理24 h组细胞侵袭数量(256.38±10.11)个、迁移相对距离(0.74±0.05),高于对照组的(185.63±23.54)个、(0.45±0.12),差异有统计学意义(t=4.783,P=0.005;t=3.864,P=0.012)。结论IL-33可通过调控NF-κB信号通路及炎症因子等的分泌,进而促进肝癌HepG2细胞增殖和迁移。
Objective To investigate the effect of IL-33 on the regulation of NF-κB signaling pathway and inflammatory factors on the proliferation and migration of HepG2 cells.Methods RT-PCR was used to detect the expression of IL-33 in different hepatocellular carcinoma cell lines(LO2,MHCC97H,LM3,HepG2,SMCC7721,Hep3B).Western Blot was used to assess the regulation of IL-33 on the activation of NF-κB signaling pathway-related proteins(NF-κB,p-NF-κB,IKB,p-IKB)in HepG2 cells.ELISA was used to measure the regulation of IL-33 on the release of inflammatory factors(IL-1α,IL-1β,IL-6,IL-18)in HepG2 cells.The EdU proliferation test was conducted to detect the effect of IL-33 on the proliferation of HepG2 cells.The CCK8 test was performed to assess the viability of HepG2 cells after 24 h and 48 h of treatment with IL-33.Transwell cell test and scratch test were conducted to examine the effect of IL-33 on HepG2 cell migration.Results The RT-PCR results showed a statistically significant difference in the relative expression of IL-33mRNA between LO2(1.01±0.01),MHCC97H(1.08±0.05),LM3(1.76±0.21),HepG2(3.88±0.35),SMCC7721(2.24±0.43),and Hep3B(2.13±0.30)(F=19.621,P=0.001).LO2 cells and MHCC97H cells had similar expression levels,while LM3,HepG2,SMCC7721,and Hep3B showed higher expression,with HepG2 cells having the highest expression.In the 24-hour group treated with IL-33,the levels of IL-1α(3185.25±194.67),IL-1β(2103.69±124.75),IL-6(446.58±36.82),and IL-18(1492.60±180.31)were higher than those in the control group(1001.58±18.65,1642.81±45.66,151.34±20.13,915.48±36.15),with statistical significance(P<0.05).The relative expression levels of phosphorylated NF-κB protein(4.53±0.36)and phosphorylated IKB protein(2.76±0.28)were higher in the 24-hour group treated with IL-33 compared to the control group(1.25±0.23,0.49±0.09),with statistical significance(P<0.05).The levels of IL-1α,IL-1β,IL-6 and IL-18 factors were higher in the 24-hour group treated with IL-33 than the control group,with a statistically significan
作者
赵盈
田明
ZHAO Ying;TIAN Ming(Tongchuan People's Hospital Anorectal,Shaanxi 727100,China;Dongzhimen Hospital of Beijing University of Chinese Medicine General surgery,Beijing 100700,China)
出处
《肝脏》
2023年第11期1331-1334,共4页
Chinese Hepatology
基金
国家自然科学基金项目(81673972)。
关键词
肝癌
IL-33
NF-ΚB信号通路
炎症
增殖
迁移
Hepatocellular carcinoma
IL-33
NF-κB signaling pathway
Inflammation
Proliferation
Migration
