Circ_SETD3 regulates gefitinib sensitivity and tumor progression by miR-873-5p-dependent regulation of APPBP2 in non-small cell lung cancer

Previous data have shown the prominent clinical efficacy of gefitinib in non-small cell lung cancer (NSCLC) patients. However, its therapeutic efficacy is limited because of the development of gefitinib resistance. This research is designed to investigate the role of circRNA SET domain containing 3, actin histidine (circ_SETD3) in the sensitivity of NSCLC to gefitinib. The expression of circ_SETD3, microRNA-873-5p (miR-873-5p) and amyloid protein-binding protein 2 (APPBP2) was detected by qRT-PCR. Protein expression was determined by western blot analysis or immunohistochemistry assay. The half-maximal inhibitory concentration of gefitinib was determined by 3-(4,5-Dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell proliferation was investigated by 5-Ethynyl-29-deoxyuridine (EdU), cell colony formation and MTT assays. Cell apoptosis was analyzed by Annexin V-fluorescein isothiocyanate and propidium iodide double staining assay. Transwell assay was employed to evaluate cell migration and invasion. Additionally, the binding relationship between miR-873-5p and circ_SETD3 or APPBP2 was predicted by starbase online database, and identified by a dual-luciferase reporter assay. Further, circ_SETD3 silencing-mediated effect on tumor sensitivity to gefitinib in vivo was confirmed by xenograft mouse model experiment. Circ_SETD3 and APPBP2 expression were upregulated, while miR-873-5p was downregulated in gefitinib-resistant NSCLC tissues and cells compared with gefitinib-sensitive NSCLC tissues or cells. Reduced expression of circ_SETD3 repressed gefitinib resistance, proliferation, migration and invasion, but induced apoptosis of gefitinib-resistant NSCLC cells. Additionally, circ_SETD3 modulated gefitinib sensitivity and tumor development by binding to miR-873-5p. APPBP2 upregulation attenuated miR-873-5p-mediated gefitinib sensitivity and NSCLC progression. Furthermore, circ_SETD3 absence improved tumor sensitivity to gefitinib in vivo. Circ_SETD3 knockdown improved gefitinib sensitivity and repressed NSCLC cell malignancy via miR-873-5p/APPBP2 axis, which provides a theoretical basis for using circ_SETD3-based therapeutic strategies to improve NSCLC sensitivity to gefitinib.</p

Table_1_Tree Size Influences Leaf Shape but Does Not Affect the Proportional Relationship Between Leaf Area and the Product of Length and Width.xlsx

The Montgomery equation predicts leaf area as the product of leaf length and width multiplied by a correction factor. It has been demonstrated to apply to a variety of leaf shapes. However, it is unknown whether tree size (measured as the diameter at breast height) affects leaf shape and size, or whether such variations in leaf shape can invalidate the Montgomery equation in calculating leaf area. Here, we examined 60 individual trees of the alpine oak (Quercus pannosa) in two growth patterns (trees growing from seeds vs. growing from roots), with 30 individuals for each site. Between 100 and 110 leaves from each tree were used to measure leaf dry mass, leaf area, length, and width, and to calculate the ellipticalness index, ratio of area between the two sides of the lamina, and the lamina centroid ratio. We tested whether tree size affects leaf shape, size, and leaf dry mass per unit area, and tested whether the Montgomery equation is valid for calculating leaf area of the leaves from different tree sizes. The diameters at breast height of the trees ranged from 8.6 to 96.4 cm (tree height ranged from 3 to 32 m). The diameter at breast height significantly affected leaf shape, size, and leaf dry mass per unit area. Larger trees had larger and broader leaves with lower leaf dry mass per unit area, and the lamina centroid was closer to the leaf apex than the leaf base. However, the variation in leaf size and shape did not negate the validity of the Montgomery equation. Thus, regardless of tree size, the proportional relationship between leaf area and the product of leaf length and width can be used to calculate the area of the leaves.</p