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 IC 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 IC 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.