Functional Study On Integrin A6β4 In Non-small Cell Lung Cancer Cells Growth

Objective:We previously knockdown integrin a6B4 expression in non-small cell lung cancer cells with RNA interference, and investigate the function of integrin a6B4 in the growth and metastasis of non-small cell lung cancer in vitro. To further provide new targets for early diagnosis and treatment of non-small cell lung cancer.Materials and methods:1. Using QRT-PCR to verify the expression of a6B4 shRNA non-small cell lung cancer cells; to observe the cellular morphology of a6B4 shRNA H460SM cells in microscope;2. Using cell counting, MTT and MTS assay to detect growth status in vitro and to study the influence of integrin a6B4 expression on the growth of non-small lung cancer cells;3. Using flow cytometry to analysis the apoptosis of a6B4 shRNA non-small cell lung cancer cells;4. We use flow cytometry to analysis the cell cycle of a6B4 shRNA non-small cell lung cancer cells, to study the function of integrin a6B4 in the process of non-small cell lung cancer cells growth.Results:1.The gene expression of H460SM-NS (non-specific cells) and that of H460SM-68,H460SM-71 (non-small cell lung cancer cells of B4 shRNA) are statistically different(P=0.000, P=0.000). The gene expression of H460SM-NS and that of H460SM-75, H460SM-76(non-small cell lung cancer cells of a6 shRNA) are also statistically different (P=0.001,P=0.000).2.The growth rate of H460SM-68,H460SM-71 (non-small cell lung cancer cells of B4 shRNA) lower than H460SM-NS (non-specific cells).There is no difference between H460SM-75, H460SM-76 and H460SM-NS.3. The apoptosis rates among non-small cell lung cancer cell lines(H460SM-NS, H460SM-68, H460SM-71, H460SM-75, H460SM-76) were not significant differen- 4. The distribution of H460SM-68 and H460SM-71 in the G0/G1 phase was significantly higher than H460SM-NS (control group), suggesting that the cell proliferation was inhibited by blocking the cell cycle in G0/G1 phase (P=0.017, P=0.013).5. PI value of H460SM-68, H460SM-71 (non-small cell lung cancer cells of B4 shRNA) is lower than that of H460SM-NS, suggesting the proliferation of shRNA-β4 transfected cells was inhibited.Conclusion:1. Non-small cell lung cancer cells that stably express a6,β4 shRNA was successfully constructed.2. Interferencing the expression of integrin B4 in non-small cell lung cancer cells could inhibit their proliferation.3. Interferencing the expression of integrin a6, B4 in non-small cell lung cancer cells might not have effect on their apoptosis.4. The possible mechinsm that B4 integrin plays an important role in non-small cell lung cancer cell growth might be related to regulation of cell cycle.