The Role And Mechanism Of CPNE1 Promoting The Proliferation And Migration Of Lung Adenocarcinoma Cells

Aim In this study,RNA interference technique was used to silence CPNE1 gene.The effect of CPNE1 gene silencing on the biological behavior of lung adenocarcinoma cell line A549 and H1299 cells was observed by in vivo and in vitro experiments,and the related mechanism was explored,which provided experimental basis for clinical treatment research.Methods Immunohistochemical method was used to detect the expression of CPNE1 protein in 128 patients with lung adenocarcinoma and 30 cases of normal lung tissue;Western blotting method was used to detect the expression of CPNE1 in lung adenocarcinoma cell strain A549,H1299,and the expression of undifferentiated bronchial epithelial cell lines.The interference efficiency of CPNE1 in the cells of A549 and H1299 was detected by qRT-PCR and Western blotting methods.Determined by CCK-8 experiment,clone formation experiment,scratches,Transwell experiment,flow cytometry detection CPNE1 gene knockout after reduction of cell proliferation and clone formation,migration,invasion ability,and the influence of the cell cycle,Using Western blotting method to detect changes in protein related to cell biology;The effect of CPNE1 gene on the tumor capacity of A549 cells was evaluated by using the nude mouse model.Results 1.The expression of CPNE1 protein in lung adenocarcinoma was significantly higher than that in normal lung tissues,and was associated with lymph node metastasis and clinical staging(P<0.05);However,there was no significant correlation between CPNE1 protein expression level and clinical pathological parameters such as age,sex and tumor size,and the difference was not statistically significant.2.Western blotting method was used to detect the expression of CPNE1 in lung adenocarcinoma cell strain A549 and H1299.The CPNE1 RNAi slow virus vector was successfully constructed,and the qRT-PCR and Western blotting tests showed that the inhibition of CPNE1 mRNA level and protein expression level of lung adenocarcinoma cells A549,H1299 was more obvious.3.The CCK-8 experiment showed that CPNE1 could inhibit the proliferation of A549 and H1299 cells after knock reduction,and the clone formation experiment showed that CPNE1 was able to inhibit the cloning of A549 and H1299 cells after the reduction of CPNE1.The scratch test and Transwell experiment showed that CPNE1 could inhibit the migration and invasion ability of A549 and H1299 cells after the reduction of CPNE1.The flow cytometry tests showed that CPNE1 could reduce the proportion of S stage cells in A549 and H1299 cell cycle after the knock reduction.Detection results of Western blotting method;Levels of related proteins,which affect cell biology,were lowered.4.The experimental results showed that the tumor capacity of A549 cells decreased after CPNE1 was decreased.Conclution 1.CPNE1 expression in lung adenocarcinoma tissues was higher than normal lung cancer,and was associated with lymph node metastasis and clinical staging.2.The CPNE1 vector of slow virus packaging can effectively reduce the expression of CPNE1 of A549 and H1299 cells.3.The CPNE1 gene can inhibit the proliferation ability of A549 and H1299 cells,clone formation ability,migration,invasion and cell cycle process,and its mechanism is related to the related gene imbalance.4.CPNE1 interference can inhibit the growth of lung adenocarcinoma in nude mice.