摘要
目的探讨黄曲霉毒素G1(AFG1)诱导的炎症反应对肺腺癌细胞增殖的影响和超氧化物歧化酶(SOD-2)高表达在其中发挥的作用。方法免疫组织化学及蛋白免疫印迹(Western Blot)方法检测AFG1诱导小鼠肺腺癌组织中增殖相关指标的表达。体外模拟AFG1诱导的炎症反应,给予AFG1、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)刺激A549细胞,Western Blot方法检测增殖相关蛋白的表达,CCK8方法及克隆形成实验检测细胞增殖能力;使用siRNA沉默SOD-2,给予上述刺激,再次检测增殖相关指标的表达情况及细胞增殖能力。结果免疫组化结果显示,AFG1诱导小鼠肺腺癌组织中增殖相关指标Survivin、B细胞淋巴瘤/白血病-xl(Bcl-xl)、增殖细胞核抗原(PCNA)和细胞周期蛋白D1(CyclinD1)均阳性表达,Western Blot结果显示以上蛋白表达均升高。体外分别给予AFG1和TNF-α+IL-6处理A549细胞,Survivin、Bcl-xl、PCNA和CyclinD1蛋白表达均无变化,CCK8和克隆形成实验结果显示细胞增殖能力没有提高。而使用siRNA转染抑制SOD-2表达后,再给予TNF-α+IL-6刺激,A549细胞增殖相关各项指标均高表达,细胞增殖能力提高。结论AFG1诱导的小鼠肺腺癌组织中增殖相关指标高表达。体外实验中AFG1和TNF-α+IL-6作用均不影响A549细胞增殖,敲低SOD-2后TNF-α+IL-6作用使A549细胞增殖能力增加,说明SOD-2是炎症反应中影响A549细胞增殖的关键指标。
Objective To investigate the effect of inflammatory microenvironment induced by aflatoxin G1(AFG1)on the proliferation of lung adenocarcinoma cells and the role of superoxide dismutase 2(SOD-2).Methods Immunohistochemistry and western blot were used to detect the expression of proliferation related indexes in lung adenocarcinoma tissues of mice induced by AFG1.The inflammatory response induced by AFG1 was simulated in vitro.A549 cells were treated with AFG1,TNF-αand IL-6 to detect the expression of proliferation related proteins.SiRNA was used to silence SOD-2,and the above stimulation was given again to detect the expression of proliferation related indicators.Results The results of immunohistochemistry showed that Survivin,Bcl-xL,PCNA,and CyclinDl were positively expressed in AFG1-induced lung adenocarcinoma.Western Blot showed that the above indicators were increased also.The expression of Survivin,BclxL,PCNA,and CyclinDl did not change in A549 treated with AFG1 or TNF-α+IL-6 in vitro.The results of CCK8 and colony formation assay showed that the cell proliferation was not improved.The expression of SOD-2 was inhibited by siRNA transfection,and then stimulated by TNF-αand IL-6;the proliferation of A549 were highly promoted.Conclusions The high expression of proliferation related markers was found in AFG1-induced lung adenocarcinoma.In vitro,AFG1 and TNF-α+IL-6 did not affect the proliferation of A549 cells,while TNF-α+IL-6 increased the proliferation of A549 cells after SOD-2 knockdown,indicating that SOD-2 might be one of the key factors that affect the proliferation of A549 cells.
作者
易力
刘宏雷
贾欣
郝增方
YI Li;LIU Hong⁃lei;JIA Xin;HAO Zeng⁃fang(Department of Pathlogy,the Second Hospital of Hebei Medical University,Shijiazhuang,Hebei 050000;Department of Neurosurgery,the Third Hospital of Shijiazhuang,Shijiazhuang,Hebei 050000,China)
出处
《热带医学杂志》
CAS
2022年第1期22-27,146,共7页
Journal of Tropical Medicine
基金
河北省卫生健康委科研基金(20190047)
关键词
黄曲霉毒素G1
炎症
超氧化物歧化酶
肺腺癌
Aflatoxin G1
Inflammation
Superoxide dismutase 2
Lung adenocarcinoma
