非小细胞肺癌FOXP3与P53突变的关联及其对Cyclin D1和CDK4的表达调控作用  

Association of FOXP3 with P53 Mutation in NSCLC and Its Regulation of Cyclin D1 and CDK4 Expression

作　　者：历春[1] 何程远 赵颖 杨红宇 LI Chun;HE Chengyuan;ZHAO Ying;YANG Hongyu(Basic Medical College of Beihua University,Jilin 132013,China;Jilin Chemical Technology Hospital,Jilin 132001,China)

机构地区：[1]北华大学基础医学院,吉林吉林132013 [2]吉林化学工业公司总医院,吉林吉林132021

出　　处：《北华大学学报（自然科学版）》2022年第6期780-784,共5页Journal of Beihua University（Natural Science）

基　　金：吉林省科技厅自然科学基金项目(YDZJ202101ZYTS089);吉林省卫生健康委员会重点实验室项目(2020J017);北华大学研究生创新项目(2021018).

摘　　要：目的探讨叉头蛋白3(FOXP3)在非小细胞肺癌(NSCLC)中的表达、与P53突变的关联及其对Cyclin D1和CDK4表达的调控作用.方法采用免疫组织化学染色法检测NSCLC组织中FOXP3和突变型P53(mt-P53)蛋白表达的相关性,应用脂质体转染法将FOXP3 siRNA瞬时转染到A549细胞中.实验分为A549组、Control-siRNA组和FOXP3-siRNA组.Real-time PCR和Western blotting方法检测FOXP3沉默效果,MTT方法检测细胞增殖情况,流式细胞术检测细胞周期情况,Western blotting方法检测Cyclin D1和CDK4的表达水平.结果NSCLC组织中,FOXP3阳性表达率为64.0%,mt-P53阳性表达率为56.0%,二者之间的表达呈显著正相关关系(r=0.342,P=0.015).与A549组和Control-siRNA组相比,FOXP3-siRNA组FOXP3 mRNA和蛋白表达水平显著下调(P<0.01),细胞增殖明显减慢(P<0.01),G_(0)/G_(1)期细胞比例明显增加(P<0.05),Cyclin D1和CDK4蛋白的表达水平显著降低(P<0.01).结论NSCLC组织中FOXP3表达与P53突变密切相关,沉默FOXP3可下调Cyclin D1和CDK4的表达,诱发细胞周期G_(0)/G_(1)期阻滞,进而抑制NSCLC细胞增殖.Objective To investigate the relationship between the expression of FOXP3 and mutant P53(mt-P53)in non-small cell lung cancer(NSCLC)and its regulation on the expression of Cyclin D1 and CDK4.Method The correlation between the expression of FOXP3 and mt-P53 proteins in NSCLC tissues was detected by immunohistochemical staining.A549 cells were transiently transfected with FOXP3 siRNA by lipofection method.Cells were divided into A549 group,Control-siRNA group and FOXP3-siRNA group.The silencing effect of FOXP3 was detected by Real-time PCR and Western blotting,the cell proliferation was detected by CCK-8,the cell cycle was detected by flow cytometry,and the expression levels of Cyclin D1 and CDK4 were detected by Western blotting.Results In NSCLC tissues,the positive expression rate of FOXP3 was 64.0%,and the positive expression rate of mt-P53 was 56.0%,and there was a significant positive correlation between the proteins(r=0.342,P=0.015).Compared with A549 group and Control-siRNA group,FOXP3 mRNA and protein expression levels in FOXP3-siRNA group were significantly down-regulated(P<0.01),cell proliferation was significantly reduced(P<0.01),and the proportion of cells in G_(0)/G_(1) phase was significantly increased(P<0.05),the expression levels of Cyclin D1 and CDK4 proteins were significantly decreased(P<0.01).Conclusion FOXP3 expression in NSCLC tissues is closely related to P53 mutation,and FOXP3 silencing can induce cell cycle arrest at G_(0)/G_(1) phase by down-regulating the expressions of Cyclin D1 and CDK4,thereby,inhibiting the proliferation of NSCLC cells.

关 键 词：非小细胞肺癌 叉头蛋白3 突变型P53 细胞周期蛋白D1 细胞周期依赖性激酶4 

分 类 号：R734.2[医药卫生—肿瘤]