Study on the effects of WASF3 antisense oligonucleotides on the proliferation and migration of non-small cell lung cancer and its mechanism

ObjectiveTo investigate the effects of antisense oligonucleotides of Wiskott-Aldrich syndrome protein family member 3(WASF3-AS) on the proliferation and migration of non-small cell lung cancer(NSCLC) cells and its mechanism.

MethodsThe expression levels of WASF3 in human NSCLC cell lung cancer cell line A549 and human normal lung cell MRC-5 were detected using RT-qPCR. The A549 cells were divide into the blank control group, WASF3-NC group(negative control group) and WASF3-AS group(antisense oligonucleotide WASF3 group). The effects of WASF3-AS on the WASF3 expression in A549 cells were detected using RT-qPCR. The CCK-8 method was used to detect the cell proliferation ability, the cloning experiment was used to detect the cell cloning ability, and the Transwell method was used to detect the cell migration ability. The mRNA and protein levels of the phosphatase and tensin homolog(PTEN), phosphatidylinositol-3-kinase(PI3K) and protein kinase B(p-Akt) in A549 cells of three groups were detected using RT-qPCR and Western blotting, respectively.

ResultsCompared with the human normal lung cells MRC-5, the mRNA expression levels of WASF3 in three group increased(P < 0.05 to P < 0.01). There was no statistical significance in the mRNA expression level of WASF3 between the blank control group and WASF3-NC group(P>0.05), but it was higher than that of WASF3-AS group(P < 0.05 and P < 0.01). Compared with the WASF3-NC group, the proliferation levels of cells in WASF3-AS group significantly decreased after 48, 72 and 96 h of transfection(P < 0.05). After 14 days of cell transfection, the clone formation rate and number of cell migration in WASF3-AS group significantly decreased compared with the blank control group and WASF3-NC group(P < 0.05). Compared with the WASF3-NC group, the expression levels of PTEN mRNA and protein in WASF3-AS group significantly increased, while the expression levels of PI3K and p-Akt mRNA and protein significantly decreased(P < 0.05).

ConclusionsThe expression level of WASF3 in human NSCLC cells is significantly higher than that in normal lung cells. The WASF3-AS may inhibit the proliferation, cloning and migration of NSCLC by regulating the PTEN-PI3K/Akt signaling pathway.