Background: Celastrol (Cela) was a natural compound that exerted anti-tumor activity in many cancer cells. Nevertheless, the molecular mechanism behind the anti-tumor role of Cela in non-small-cell lung carcinoma (NSCLC) remains to be clarified.
Methods: Flow cytometry was used to analyze cell cycle progression and apoptosis. Colony formation assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay were used to analyze cell proliferation. Cell migration and invasion abilities were assessed by transwell assays. Quantitative real-time polymerase chain reaction (qRT-PCR) was implemented for the detection of RNA levels. Western blot assay was used for the determination of protein levels. Dual-luciferase reporter assay was conducted to confirm the interaction between microRNA-33a-5p (miR-33a-5p) and circular RNA SATB homeobox 2 (circ_SATB2) or E2F transcription factor 7 (E2F7). Xenograft tumor assay was conducted to test the roles of Cela and circ_SATB2 in NSCLC progression in vivo.
Results: Cela hampered the malignant behaviors of NSCLC cells. Cela down-regulated circ_SATB2 level in NSCLC cells. Cela stimulation-induced suppressive influence in NSCLC progression was alleviated by circ_SATB2 accumulation. E2F7 interference overturned circ_SATB2-mediated effects in Cela-stimulated NSCLC cells. MiR-33a-5p was a target of circ_SATB2, and E2F7 was verified as a target of miR-33a-5p. Circ_SATB2 attenuated Cela-mediated effects through targeting miR-33a-5p in NSCLC cells. Cela-mediated suppressive effect on tumor growth was partly attenuated by the overexpression of circ_SATB2 in vivo.
Conclusion: Cela suppressed NSCLC development through regulating circ_SATB2/miR-33a-5p/E2F7 signaling cascade.
Keywords: NSCLC, celastrol, circ_SATB2, miR-33a-5p, E2F7