| Objective:Lung cancer is the fastest growing morbidity and mortality rate,and it is the most serious malignant tumor to people's health and life.It seriously affects people's quality of life and life expectancy.Most lung cancer patients have been diagnosed at an advanced stage.This is closely related to the failure to find an early diagnosis of lung cancer.Therefore,lung cancer patients should be diagnosed as early as possible so that patients can seek medical treatment in time and reduce the risk of lung cancer.Finding molecular markers that predict the occurrence of lung cancer is crucial,so it is an urgent task to find an early diagnosis of lung cancer.As a member of the miRNA family,miR-375 has now been shown to participate in the basic processes of migration,proliferation,metastasis,differentiation,and apoptosis of various tumor cells.Several experimental results have shown that miRNA-375 is expected to become a new target for cancer therapy and drug treatment,and may become an important biomarker for tumor diagnosis and prognosis.In this study,miRNA-375 transfected by Lentivirus in lung cancer A549 cells was designed to investigate its role in epithelial mesenchymal transformation of lung cancer cells,and the invasion and migration of miRNA-375 through lung cancer cells via CIP2 A signaling pathway.,proliferation,and apoptosis.Methods:The experiments were divided into three groups: A549 cells stably over-expressing miRNA-375 were used as the experimental group,and A549 cell line transfected with empty vector was used as a negative control group.The A549 cell line without any treatment was used as a blank control.The expression of mi RNA-375 in three groups of A549 cells was detected by real-time fluorescence quantitative PCR.Western blot was used to detect E-cadherin,a protein marker related to epithelial mesenchymal transformation in three groups of A549 cells.E-cadherin,vimentin,and expression levels of CIP2 A,MYC,AKT,P-AKT,and other related proteins in three groups of cells;three groups of cells detected by Trans-well chamber model test and scratch healing test Theability of cell invasion,metastasis and migration in A549 cells was detected by CCK8 technique.The proliferation of A549 cells was detected by flow cytometry.Results:The results of real-time fluorescence quantitative PCR showed that the expression level of miRNA-375 was significantly up-regulated in the experimental group compared with the negative control group and the blank control group(P<0.05),but there was no significant difference between the negative control group and the blank control group(P>0.05).,to prove successful transfection.Western blot results showed that A549 cells in the experimental group were characterized by mesenchymal cells in their cytoskeleton and morphology after transfection with lentivirus,and the expression of E-cadherin marker protein was decreased,while Vimentin expression was up-regulated and statistically analyzed.Significance(P<0.05),but there was no significant difference between the negative control group and the blank control group(P>0.05);Trans-well chamber model and scratch test showed that the experimental group was relative to the negative control group and the blank control group A549 cells The invasion and metastasis migration ability was enhanced and statistically significant(P<0.05),but there was no significant difference between the negative control group and the blank control group(P>0.05);Western blot results showed CIP2 A,MYC,P-AKT was highly expressed in the experimental group and statistically significant(P<0.05),AKT,?-actin expression in the three groups was not statistically significant(P>0.05);CCK8 technical results showed that the cell proliferation ability in the experimental group Strong and statistically significant(P<0.05),but there was no significant difference between the negative control group and the blank control group(P>0.05);The results of flow cytometry showed that apoptosis was less and statistically significant in the experimental group(P<0.05),while there was no significant difference between the negative control group and the blank control group(P >0.05).Conclusion:1.Abnormally high expression of microRNA-375 promotes the occurrence and development of epithelial mesenchymal transition in lung cancer A549 cells.2.microRNA-375 may promotes the invasion,metastasis,migration and proliferation of lung cancer A549 cells via the CIP2 A pathway and inhibits its apoptosis.3.microRNA-375 is expected to become a new target for the diagnosis and treatment of lung cancer,and may become an important biomarker for early diagnosis and prognosis of the disease.Significance:In this study,a large amount of literature was consulted and through further research in this experiment,miR-375 was found to be abnormally expressed in lung cancer and involved in the regulation of tumor cells.This not only relates to mesenchymal transition of epithelial cells but also promotes lung cancer by modulating CIP2 A pathway.Invasion,metastasis,migration and proliferation of A549 cells inhibit their apoptosis.miRNA-375 is expected to become a new target for the treatment of lung cancer,and may become an important biomarker for disease diagnosis and prognosis.This study is a challenging scientific attempt and will enrich the regulatory mechanisms of tumor invasion,migration,and proliferation. |
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