摘要
目的研究在非小细胞型肺癌(non-small cell lung cancer,NSCLC)发展进程中,p53通过调控人成纤维细胞生长因子13(fibroblast growth factor 13,FGF13)的表达对细胞增殖的影响,并探讨其相关分子机制。方法以人正常肺上皮细胞BEAS-2B和NSCLC A549细胞为研究对象,对A459细胞进行单独干扰p53和FGF13后,采用CCK8法检测对细胞增殖的影响;qPCR和Western blot法检测p53干扰后对相关因子和蛋白表达的影响;流式细胞术检测对细胞周期的影响。进一步采用p53和FGF13联合干扰,验证对A549细胞增殖的影响。结果p53干扰后,A549细胞增殖明显加快(P<0.01),细胞周期抑制因子p21蛋白表达水平显著降低(P<0.01),CyclinE和FGF13蛋白表达水平显著升高(P均<0.01)。FGF13表达降低后,G1期细胞数量升高14%,S期细胞数量降低10%;p21基因mRNA转录水平显著升高,CDK2基因mRNA转录水平显著下降,而细胞周期蛋白CyclinE表达水平明显降低(P均<0.01)。超表达FGF13后,p21基因mRNA转录水平明显降低,而CyclinE蛋白表达水平上调3倍(P均<0.01),表明FGF13在A549细胞中发挥促增殖的作用。进一步的单干扰或联合干扰表达证明,p53通过抑制FGF13的表达降低CyclinE蛋白的表达。结论FGF13作为一个促癌因子,受到肿瘤抑制蛋白p53的直接调控,通过细胞周期抑制因子p21通路,共同影响A549细胞周期中G1/S检控点。
Objective To investigate the effect of p53 on cell proliferation by regulating the expression of human fibroblast growth factor 13(FGF13)in development of non-small cell lung cancer(NSCLC)as well as the relevant molecular mechanism.Methods Normal human lung epithelial cell line BEAS-2B and NSCLC cell line A549 were used as objects.After interference with p53 and FGF13 alone,the effect on proliferation of A459 cells was detected by CCK8 method.The effects of p53 interference on expressions of related factors and proteins were evaluated by qPCR and Western blot respectively,while that on cell cycle by flow cytometry.The effect of combined interference with p53 and FGF13 on A549cell proliferation was further verified.Results The proliferation of A549 cells increased significantly after p53 interference(P<0.01),while the expression level of cell cycle inhibitor p21 decreased significantly(P<0.01),and those of CyclinE and FGF13 increased significantly(P<0.01).After the decrease of FGF13 expression level,the number of cells in G1 phase increased by 14%,while that in S phase decreased by 10%;the transcription level of p21 mRNA increased significantly,while that of CDK2 mRNA decreased significantly;however,the expression level of CyclinE protein decreased significantly(each P<0.01).After overexpression of FGF13,the transcription level of p21 decreased significantly,while the expression level of CyclinE protein increased by 3 folds(each P<0.01),indicating that FGF13promoted the proliferation of A549 cells.Furthermore,single interference or combined interference showed that p53decreased the expression of CyclinE protein by inhibiting the expression of FGF13.Conclusion As a tumor-promoting factor,FGF13 is directly regulated by tumor suppressor protein p53,which jointly affects the checkpoint of G1/S in A549cell cycle through cell cycle inhibitor p21 pathway.
作者
赵倩文
唐铖铖
雷静静
张欣然
李豆豆
曾文先
路宏朝
ZHAO Qian-wen;TANG Cheng-cheng;LEI Jing-jing;ZHANG Xin-ran;LI Dou-dou;ZENG Wen-xian;LU Hong-zhao(School of Biological Science Technology and Engineering,Shaanxi University of Technology,Hanzhong 723001,Shaanxi Province,China)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2022年第5期567-574,共8页
Chinese Journal of Biologicals
基金
国家级大学生创新创业训练项目(201910720011)。
