Study On The Inhibition Of Mir-16-1on Biological Behavior Of A549Cell Lines And Arid1A Expression In Non-small Cell Lung Cancer

Part1:The inhibition of miR-16-1on biological behavior of the A549cell linesObjective:To study the miR-16-1expression in non-small cell lung cancer (NSCLC) and the inhibition of miR-16-1on A549cell lines, which provide a basis for further studying the relationship between the miR-16-1and NSCLC.Methods:We detected the expression of miR-16-1in40matched NSCLC and adjacent non-tumor lung tissues by qRT-PCR. The miR-16-1expression was compared in NSCLC tissues with in adjacent non-tumor tissues, and the relationship of expression was investigated between NSCLC and miR-16-1.In order to investigate the biological behaviors of the miR-16-1on A549cell lines, miR-16-1mimics with biological activity in vivo were transfected into A549cells with lipofectamine2000. The expression of miR-16-1was measured by qRT-PCR. The proliferation of cell was measured by MTT assay. Cells were stained with a combination of Annexin V-FITC and propidium iodide (PI), and the apoptosis cells was analyzed by the flow cytometry system. PI stained was used to calculate the cell cycle by the flow cytometry. The ability of invasiveness was measured by transwell assay.Results:The results showed the expression of the miR-16-1in lung cancer tissues was significantly lower than that in adjacent non-tumor tissues from40cases of NSCLC. The expression of miR-16-1in lung cancer tissue was an average of only0.34times compared with its adjacent non-tumor tissue, which difference was statistically significant, p<0.05. Overexpression of miR-16-1by transfection with hsa-miR-16-1mimics induced cell cycle arrested in G0-G1. The rate of apoptosis significantly increased in A549after transfection. The invasiveness and proliferation activity of A549cells were significantly lower after transfection.Conclusion:These data indicate that miR-16-1may act as a tumor suppressor on growth, proliferation and invasion in A549, and it may be a potential therapeutic target for NSCLC. Part2:Study on ARID1A expression in non-small cell lung cancerObjective:To study ARID1A expression in A549cell lines and tissues of non-small cell lung cancer (NSCLC), and investigate the mechanism of action between ARID1A and NSCLC.Methods:A549cell lines and40cases of NSCLC specimens were adopted. The immunofluorescence and confocal laser microscopy were applied to detect and localize ARID1A expression in A549cell line. We detected the mRNA expression of ARID1A in40matched NSCLC and adjacent non-tumor lung tissues by qRT-PCR. Western blotting and immunohistochemistry were used to analyze the ARID1A expression in order to investigate the relationship between ARID1A expression and the clinical as well as pathological features of NSCLC, and its clinical significance was investigated in NSCLC.Results:The results showed that the ARID1A protein located specifically in the nucleus by immunofluorescence and confocal microscopy, the positive expression of ARID1A in A549cell line was definite. The mRNA expressions of the ARID1A in lung cancer tissues were low expressions with an average of only0.63times compared with matched adjacent non-tumor tissues, which difference was statistically significant, p<0.05. Western blotting and immunohistochemistry showed that the expressions of ARIDA protein in cancer tissues were lower than that in their adjacent non-tumor tissues. The ARID1A positive expressions mainly were found in the early periods and limited to stage â… , â…¡, and no positive expression was found in the organization with distant metastasis. The positive expression of ARID1A had no difference in gender, histological type and lymph node metastasis.Conclusion:ARID1A is a frequent loss of expression acts as a tumor suppressor gene in NSCLC, which occurs in the advanced lung cancer, loss of expression of ARID1A has a relationship with progression of lung cancer.