Effect Of Nm23-H1 Gene Transfection Through Recombinant Adenoviral Vector Containing Nm23-H1 On Proliferation And Colony Formation Of Human Lung Adenocarcinoma Cells

Objective :To explore the role of nm23-H1 gene on differentiation , proliferation and colony formation of high metastatic human lung adenocarcinoma cells, to check the role of nm23-H1 to malignant tumor,thus provide some theory and method basis for gene therapy of oral and other carcinoma by nm23-H1 in later. Methods: A recombinant adenoviral containing nm23-H1 was transfected into lung adenocarcinoma ceils Anip973.Test such as immunohistochemistry,immunofluorescence and flow cytometry were used to analyze the expression levels of nm23-H1, MTT were used to analyze the proliferation of the tumor through cells curve of growth (P< 0.05), and flow cytometry was used to test the changes of cell cycle, apoptosis befoe and after gene transfection, The tumor formation was tested by colony-forming in soft-agarose (P<0.05) .Results: After amplification, the virus titer reached to 10⁷.At the time MOI=80, the transfection efficiency could more than 80%. When transfection successfully,the nm23-H1 protein expression was upregulated significantly. The ratio of G₀/G₁ and S period was degrade, G₂/M and apoptosis was raised.The ability of proliferation and tumor formation were remarkably decreased.Conclusion: 1.Through Ad, after transfected with rhAd-nm23-H1,the expression of nm23-H1 in human lung adenocarcinoma Anip973 cells can significantly improved ; 2. After transfected,the transition from G₂-phase to M-phase of Anip973 cells was obviously suppressed, and apoptosis is induced; 3. After transfected,up-regulation of nm23-H1 could suppress the ability of proliferation and colonies formation in human lung adenocarcinoma Anip973 cells in vitro.