摘要
目的探讨水通道蛋白1(AQP1)基因的过表达对A549人肺腺癌细胞增殖的影响及可能机制。方法构建和包装AQP1慢病毒过表达载体,感染A549细胞,实时荧光定量PCR检测AQP1 mRNA水平,Western blot法检测AQP1蛋白水平。在A549细胞过表达AQP1后,噻唑蓝(MTT)实验检测细胞增殖,细胞克隆形成实验检测细胞克隆形成能力,并以Western blot法检测细胞β联蛋白(β-catenin)磷酸化水平。结果成功构建AQP1过表达慢病毒载体,病毒滴度为(2.87±0.54)×10~8 IU/mL;慢病毒载体感染A549细胞后,细胞AQP1 mRNA和蛋白水平显著增加;细胞增殖能力显著提高,细胞克隆形成数显著上升;β-catenin总蛋白水平增加,但β-catenin磷酸化显著降低。结论过表达AQP1促进A549细胞增殖并抑制β-catenin的磷酸化。
Objective To investigate the effect of over-expression of aquaporin 1(AQP1)gene on the proliferation of human lung adenocarcinoma A549 cells and its possible mechanism.Methods A lentiviral vector for over-expression of AQP1 was constructed,identified and used to infect A549 cells.Real-time quantitative PCR and Western blot analysis were performed to detect the expression of AQP1 mRNA and protein,respectively.In the AQP1-over-expressing A549 cells,MTT assay,clone formation assay and Western blot analysis were used to assess cell proliferation,cell clone forming ability,andβ-catenin phosphorylation level,respectively.Results The retroviral expression vector of AQP1 with a virus titer of(2.87±0.54)×10~8 IU/mL was obtained and A549 cells were infected with it to get a stable AQP1 over-expressing cell line.In this cell line,the AQP1 mRNA and protein levels were significantly raised,the cell proliferation ability was significantly improved,and the number of cell clones was significantly elevated.The total protein level ofβ-catenin increased,but the phosphorylation ofβ-catenin decreased significantly.Conclusion Over-expression of AQP1 promotes A549 cell proliferation and inhibitsβ-catenin phosphorylation.
作者
王会峰
董辉
吴媛媛
乔立娇
金向明
WANG Huifeng;DONG Hui;WU Yuanyuan;QIAO Lijiao;JIN Xiangming(Department of Oncology,Cancer Hospital,General Hospital of Ningxia Medical University,Yinchuan 750000,China;Center of Research Equipment Management,General Hospital of Ningxia Medical University,Yinchuan 750000,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2020年第5期413-418,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
宁夏自然科学基金(2019AAC03197)。
