摘要
目的探讨敲低TM4SF1对PI3K/AKT通路及鼻咽癌细胞转移能力的影响。方法通过对鼻咽癌细胞HONE1转染siNC和siTM4SF1,将HONE1细胞分为正常对照组和TM4SF1敲低组,用qRT-PCR和Western blot验证TM4SF1敲低效率,采用划痕修复实验和转移小室(Transwell)检测TM4SF1敲低后HONE1细胞的转移能力,Western blot检测TM4SF1敲低后HONE1细胞的AKT、p-AKT、E-cad、N-cad、Vimentin和MMP2蛋白水平。随后使用AKT特异性抑制剂MK2206(100 nmol/L)和siTM4SF1处理HONE1细胞,将HONE1细胞分为正常对照组、MK2206组以及siTM4SF1+MK2206组,采用Transwell实验检测HONE1细胞的转移能力。结果与正常对照组相比,TM4SF1敲低组的TM4SF1蛋白及mRNA表达水平显著降低(P<0.05),并且TM4SF1敲低组HONE1细胞转移能力明显下降(P<0.05),p-AKT、N-cad、Vimentin和MMP2蛋白水平明显下降(P<0.05),E-cad蛋白水平明显升高(P<0.05)。与正常对照组相比,MK2206组HONE1转移能力显著降低(P<0.05),且siTM4SF1+MK2206组相比于MK2206组转移能力进一步降低(P<0.05)。结论敲低TM4SF1可以通过PI3K/AKT通路抑制鼻咽癌细胞增殖和转移能力。
Objective To investigate the effect of TM4SF1 knockdown on the PI3K/AKT pathway and nasopharyngeal carcinoma cell metastasis.Methods HONE1 cells were transfected with siNC and siTM4SF1,respectively,named as control group and TM4SF1 knockdown group.The knockdown efficiency was verified by qRT-PCR and Western blot.The metastasis ability of HONE1 cell line was detected by Transwell and wound healing assay.Meanwhile,the protein levels of AKT,p-AKT,E-cad,N-cad,Vimentin and MMP2 were detected by Western blot.Subsequently,the HONE1 cells were divided into control group,MK2206(an AKT specific inhibitor)group and siTM4SF1+MK2206 group,and the metastasis ability was detected by Transwell assay.Results Compared with control group,the protein and mRNA levels of TM4SF1 were significantly decreased in TM4SF1 knockdown group(P<0.05),and the metastasis ability of HONE1 cells was decreased(P<0.05).The protein levels of p-AKT,N-cad,Vimentin and MMP2 were significantly downregulated after TM4SF1 knockdown(P<0.05),while the protein level of E-cad was significantly upregulated(P<0.05).Compared with control group,the metastasis ability of HONE1 in MK2206 group was significantly decreased(P<0.05).In addition,the metastasis ability of HONE1 in siTM4SF1+MK2206 group was significantly lower than that in MK2206 group(P<0.05).Conclusion Knockdown of TM4SF1 can inhibit the nasopharyngeal carcinoma cell metastasis by targeting the PI3K/AKT pathway.
作者
陈慧
魏柏
CHEN Hui;WEI Bo(Department of Haematology,Liyuan Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430077,China;Department of Oncology,Liyuan Hospital,Tongji Medical College,Huazhong University of Science and Technology)
出处
《山西医科大学学报》
CAS
2020年第5期397-402,共6页
Journal of Shanxi Medical University
基金
湖北省自然科学基金面上项目(2019CFB706)。
作者简介
陈慧,女,1983-04生,硕士,住院医师,E-mail:255907215@qq.com;通讯作者:魏柏,E-mail:doctorwb@hust.edu.cn。
