Abstract:
Objective To investigate the reversal effect of TIP30 on gefitinib-resistant cells in non-small cell lung cancer (NSCLC) and explore its possible mechanisms.
Methods TIP30 was up-regulated in gefitinib resistance of NSCLC PC9/GR cells transfected with lentiviral vector LV-TIP30.PC9/GR, PC9/GR-LVTIP30 and PC9/GR-LVNC cells were as research objects. Each group was randomly assigned into six groups and treated with or without 5 μmol/L gefitinib. CCK8 was used to detect the inhibition rate of cell proliferation, wound healing assay was used to test the cell migration ability, transwell assay was used to detect the cell invasion ability, Western blot experiment was used to detect p-MEK, p-ERK, p-AKT and nuclear EGFR protein expression levels.
Results In the non drug treatment group, the proliferation, migration and invasion ability of PC9/GR-LVTIP30 cells were significantly inhibited compared with PC9/GR cells (P < 0.05), and no significant difference was found between PC9/GR-LVNC and PC9/GR cells. The inhibitory effect of PC9/GR-LVTIP30 cells in gefitinib treatment group was more obvious than that of PC9/GR cells (P < 0.05). After overexpression of TIP30, the expression level of p-MEK, p-ERK, and p-AKT was decreased. The expression level of nuclear EGFR was lower than that of PC9/GR, and the difference was statistically significant (P < 0.05).
Conclusions TIP30 upregulation can inhibit the proliferation, migration and invasion of gefitinibresistant cell line PC9/GR of human NSCLC, and it can reverse the resistance of PC9/GR to gefitinib, partially restore PC9/GR cells' sensitivity to gefitinib. The mechanism may correlate with the inhibition for the activation of nuclear-internalized EGFR downstream signaling pathway-related proteins p-AKT, p-ERK, and p-MEK.
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