This study aimed to investigate the expression, function, and possible mechanism of Src in the Hep-2 cell line. We used Src-specific small interfering RNA (siRNA) to inhibit the expression of Src in Hep-2 cells. RT-PCR and Western blot were applied to evaluate the expression level of Src after RNA interference, and the MTT assay and flow cytometry were used to observe the expression of PI-3 K and Akt. siRNA can downregulate the expression of Src in Hep-2 cells. Downregulation of Src decreased PI-3 K and Akt expression. We found that Src knockdown inhibits the proliferation of Hep-2 cells and the growth of laryngeal carcinoma in vivo. This study has demonstrated that Src participates in the regulation of apoptosis through the Src/PI-3 K/Akt signaling pathway in the Hep-2 cell line. Silencing of Src by siRNA is a viable approach in laryngeal carcinoma treatment.