建立稳定敲减TNIK表达的A549细胞系及对其初步分析  

Establishment and Preliminary Analysis of Cell Line A549 with Stable Knockdown of TNIK Expression

作　　者：孙雪花 尹讯 王茹 张涛[1,2] 陈全 张春冬[1,2] SUN Xuehua;YIN Xun;WANG Ru;ZHANG Tao;CHEN Quan;ZHANG Chundong(Department of Biochemistry and Molecular Biology,Chongqing Medical University,Chongqing 400016,China;Department of Immunology,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学生物化学与分子生物学教研室,重庆400016 [2]重庆医科大学分子医学与肿瘤研究中心,重庆400016 [3]重庆医科大学免疫学教研室,重庆400016

出　　处：《中国细胞生物学学报》2022年第3期421-428,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:82173243);重庆市科委自然科学基金(批准号:cstc2021jcyjmsxmX0190)资助的课题。

摘　　要：TNIK(TRAF2 and NCK interacting kinase)属丝氨酸/苏氨酸激酶家族成员,在多种生理及病理过程中起关键作用。研究发现,TNIK在肺鳞癌组织中高表达且可驱动癌细胞增殖等恶性表型,但其在肺腺癌中的作用仍未知。该研究在构建稳定敲减TNIK表达的肺腺癌A549细胞后,流式细胞术检测发现稳定敲减TNIK阻滞细胞周期进程并诱发凋亡,细胞增殖及运动实验证明肺癌细胞增殖与迁移能力被显著抑制。进一步通过免疫荧光染色分析发现,稳定敲减TNIK表达后会诱导细胞微丝骨架排列紊乱并抑制黏着斑动态周转(组装/解聚)。综上研究结果表明,肺腺癌细胞中TNIK可能通过调节微丝骨架排列,从而影响黏着斑动态周转最终控制细胞增殖及迁移运动;推测肺腺癌细胞中高表达的TNIK可能通过调控细胞微丝骨架系统以维持癌细胞恶性表型。TNIK (TRAF2 and NCK interacting kinase),a member of the family of serine/threonine kinase,has played a key role in many physiological and pathological process.Recent studies have found that TNIK is highly expressed in lung squamous cell carcinoma tissues and drives cancer cell proliferation and other malignant phenotypes,but its role in lung adenocarcinoma is still poorly understood.In this study,a lung adenocarcinoma A549 cell line with stable knockdown TNIK expression were constructed.Flow cytometry showed that the cell cycle was arrested and apoptosis was induced.Cell proliferation and movement experiments proved that the proliferation and migration of lung cancer cells were significantly inhibited.Further immunofluorescence staining analysis showed that stable knockdown of TNIK expression could induce disorder of cell actin cytoskeleton and inhibit dynamic turnover (assembly/depolymerization) of focal adhesions.In conclusion,TNIK in lung adenocarcinoma cells may influence the dynamic turnover of focal adhesions and ultimately control cell proliferation and migration by regulating the arrangement of actin cytoskeleton.It is speculated that the highly expressed TNIK in lung adenocarcinoma cells can maintain the malignant phenotype of cancer cells by regulating the actin cytoskeleton system.

关 键 词：肺腺癌细胞 细胞增殖 细胞运动 微丝骨架 TNIK 

分 类 号：R734.2[医药卫生—肿瘤]