Objective: Lymphovascular invasion (LOI), a key pathological feature of head and neck squamous cell carcinoma (HNSCC), predicts poor survival. However, the associated clinical characteristics remain uncertain, and the molecular mechanisms are largely unknown. Methods: Weighted gene co-expression network analysis was performed to construct gene co-expression networks and investigate the relationship between modules and LOI clinical trait. Functional enrichment and KEGG pathway enrichment analysis were performed for differentially expressed genes using DAVID database. The protein-protein interaction network was constructed using Cytoscape software, and module analysis was performed using MCODE. Prognosis role and expression analysis was further validated by survival analysis, GEPIA analysis and HPA database. Multivariable Cox regression analysis was used to establish a prognostic risk formula and the areas under the receiver operating characteristic curve (AUCs) were used to evaluate prediction efficiency. And the potential targeted LOI molecular agents were identified with DrugBank. Results: 10 co-expression modules in two key modules (turquoise and pink) associated with tumor LOI were identified. Functional enrichment and KEGG analysis identified turquoise and pink modules played significant roles in the progression of HNSCC. The seven hub genes (CNFN, KIF18B, KIF23, PRC1, CCNA2, DEPDC1 and TTK) in two modules were identified and validated by survival analysis and expression analysis. Multivariable Cox regression analysis was used to establish a prognostic risk formula (risk score = EXP DEPDC1 * 0.32636 + EXP CNFN * (-0.07544). The low-risk group had a better OS than the high-risk group ( P <0.001), and the areas under the receiver operating characteristic curve (AUCs) of 1-, 3- and 5-year OS were 0.582, 0.634 and 0.636, respectively. Eight small molecular agents, including XL844, AT7519, AT9283, Alvocidib, Nelarabine, Benzamidine, L-Glutamine, and Zinc, may be a candidate drug for treating LOI ( P < 0.05). Conclusions: Our research revealed the two-mRNA signature could serve as an independent biomarker to predict LOI risk, which provide some new insights into LOI of HNSCC. Additionally, the small molecular agents may be a candidate drug for treating LOI.