Cucurbitacin B enhances apoptosis in gefitinib resistant non-small cell lung cancer by modulating the miR-17-5p/STAT3 axis

Abstract

Tyrosine kinase inhibitors, such as gefitinib, are currently widely used as targeted therapeutics for non-small cell lung cancer (NSCLC). Although drug resistance has become a major obstacle to successful treatment, mechanisms underlying resistance to gefitinib remain unclear. Therefore, the present study aimed to investigate the impact of adjunc- tive cucurbitacin B (CuB) on gefitinib resistance (GR) in the PC9 cell line, including identifying underlying mechanisms. Reverse transcription-quantitative PCR demonstrated signifi- cant downregulation of microRNA (miR)-17-5p expression in GR PC9 cells (PC9/GR), and this could be reversed by CuB. During combination treatment with CuB and gefitinib at IC25, PC9/GR cell proliferation was downregulated, and apoptosis was upregulated. The presence of a miR-17-5p inhibitor negated the effects of CuB and gefitinib, whereas the pres- ence of a miR-17-5p mimic enhanced them. Luciferase assays demonstrated that the hypothetical target gene, signal trans- ducer and activator of transcription 3 (STAT3), was directly targeted by miR-17-5p. Moreover, significant elevation of the STAT3 protein and phosphorylation levels in PC9/GR cells was reversed by the addition of CuB, despite a lack of change in STAT3 transcription level. During combined treatment with CuB and gefitinib at IC25, the STAT3 protein expression was negatively associated with the expression of miR-17-5p. Overexpression of STAT3 increased proliferation and decreased apoptosis and the protein levels of apoptosis-related factors cleaved caspase-3 and cleaved caspase-9 of PC9/GR cells. Findings indicated that STAT3 protein and phosphoryla- tion levels became elevated in response to gefitinib, and that CuB-induced miR-17-5p expression led to STAT3 degradation, thereby ameliorating GR. In summary, CuB reduced the prolif- eration of GR PC9 cells by modulating the miR-17-5p/STAT3 axis, and may represent a promising potential novel strategy for the reversal of GR.