Scutellarein treats neuroblastoma by regulating the expression of multiple targets

Abstract

The aim of this study is to investigate the effect of scutellarein on the proliferation of neuroblastoma cells and the underlying mechanism. Six cell lines were used with drug intervention. Cell Counting Kit-8 was used to select the best, namely, SH-SY5Y, and then its IC₅₀ value was determined. To further investigate the mechanism of scutellarin affecting SH-SY5Y proliferation, quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression levels of 11 factors. Scutellarin administration with 300 μM significantly reduced the number of SH-SY5Y, especially on the 3rd day of exposure to scutellarin. The IC₅₀ value of scutellarin in SH-SY5Y cells was determined to be 117.8 μM. But the practical results showed that 300 μM was the optimal concentration of scutellarin. qRT-PCR further detected upregulated maternally expressed gene 3 (MEG3), oncogene c-Fos (c-FOS), and c-jun and downregulated M2 isoform of pyruvate kinase (PKM2), non-SMC Condensin I Complex Subunit H (NCAPH), epidermal growth factor receptor (EGFR), transforming growth factor (TGF)-β1, and TGF-α, suggesting that scutellarin with 300 μM volume inhibited the survival of SH-SY5Y by regulating the expression of these 8 factors. Scutellarin could be a novel drug for the treatment of neuroblastoma, and its underlying mechanism may be related to the upregulated levels of MEG3, c-FOS, and c-jun and downregulated the expression of PKM2, NCAPH, EGFR, TGF-β1, and TGF-α.

Keywords: HS‐SY5Y; neuroblastoma; proliferation inhibition; scutellarin.

Figure 1

Molecular formula of scutellarin. [Color figure can be viewed at wileyonlinelibrary.com]

Figure 2

Cellular screening. (A) Morphology of six alternative cell lines in five groups with different drug dosages (Control, Scu‐35, Scu‐75, Scu‐150, and Scu‐300) under the light field. (B) The bar chart shows the inhibition ratio of each cell line in each group by Cell Counting Kit‐8 (*p < 0.05) (Control: no scutellarin; Scu‐75: add 75 μM scutellarin; Scu‐150: add 150 μM scutellarin; and Scu‐300: add 300 μM scutellarin). [Color figure can be viewed at wileyonlinelibrary.com]

Figure 3

The best‐fit values of scutellarin concentration. The curve chart reveals the dose–response curve following medication. The table in the top right corner provides the best‐fit values of IC₅₀.

Figure 4

Morphology of SH‐SY5Y after scutellarin administration. SH‐SY5Y were divided into four groups (Control: no scutellarin; Scu‐75: Add 75 μM scutellarin; Scu‐150: Add 150 μM scutellarin; and Scu‐300: Add 300 μM scutellarin) and were cultured for 3 days. The morphology in each group of each day was visible under the light field. [Color figure can be viewed at wileyonlinelibrary.com]

Figure 5

Molecular changes following scutellarin administration. The bar charts (A–K) reveal the changes in the relative expressions of c‐FOS, c‐jun, EGF, EGFR, IL‐6, MEG3, NCAPH, PKM2, Survivin, TGF‐α, and TGF‐β1 driven by scutellarin administration (A–K). c‐Fos, oncogene c‐Fos; EGF, epidermal growth factor; EGFR, epidermal growth factor receptor; IL‐6, interleukin‐6; MEG3, maternally expressed gene 3; NCAPH, non‐SMC Condensin I Complex Subunit H; PKM2, M2 isoform of pyruvate kinase; TGF‐α, transforming growth factor‐α; TGF‐β1, transforming growth factor‐β1.

Figure 6

Scutellarein regulates the proliferation of SH‐SY5Y by multiple targets. [Color figure can be viewed at wileyonlinelibrary.com]