B-219 Apoptotic effect and cell cycle arrest by Juniperus communis extract on oral cancer

Background According to the American Cancer Society, oral cancer accounted approximately 59,660 new cases of oral cavity or oropharyngeal cancer and 12,770 deaths from oral cavity or oropharyngeal cancer in 2025. The five-year survival rate of oral cancer patients decreases significantly as the stage increases. The recurrence rate of oral cancer within three years is as high as 86%, accompanied by high metastasis ability and drug resistance. The treatment for oral cancer mainly includes surgical resection combined with chemotherapy or radiotherapy. However, the most commonly used chemotherapy drugs are prone to serious side effects. For example, 5-Fluorouracil (5-FU) can cause diarrhea, vomiting, hair loss, and low immunity in patients. It is urgent need to develop new drugs that can effectively inhibit cancer growth and reduce side effects. In this study, we combined use of Juniperus communis crude plant extract (JCE) with clinical drug 5-FU against oral cancer cell lines. Method The juniper berries are dried and ground, and the powder is placed in a distillation column. Extraction at 100-105°C at a flow rate of 7.2 ml/min 100 minutes, the effective ingredients are taken out by steam, and then the required juniper fruit crude extract is separated through the condensation system. JCE and 5 Fluorouracil (5 FU; Sigma) were dissolved in DMSO. The effect of JCE on the viability of OECM1, SCC-25, SVEC, and MDCK was measured using an MTT assay. Flow cytometry was used to analyze the cell cycle distribution and cell death. The protein expression in oral cancer cells after treatment with JCE or 5-FU was evaluated by western blot analysis. Results The results showed that JCE inhibited the growth of oral cancer cells and synergistic inhibitory effect when combined with 5-FU. It also induced cell cycle arrest, increased the percentage of subG1 cells and activated intracellular caspases, thereby promoting apoptosis and leading to cancer cell death. Compared with a single drug, JCE+5-FU more significantly regulated the expression of AKT/mTOR, Cyclins/CDKs and Caspase 3, 8, 9 proteins, and exhibited a better tumor suppressor effects. More importantly, JCE attenuated the development of drug resistance in oral cancer cells treated with 5-FU over a long period of time. This project confirmed the synergistic inhibitory effects and mechanism of JCE combined with 5-FU in oral cancer cells. In the future, it will be further developed as an anticancer drug or adjuvant for the treatment of oral cancer or used in combination with clinical drugs. Conclusion The results of this study indicate that JCE and 5 FU, when used in combination, can inhibit cell growth by inhibiting the Akt/mTOR/p70S6K pathway and cell cycle-related molecules, while activating apoptotic proteins to induce oral cancer cell death. JCE combines the clinical drug 5-FU with the tumor suppressor and anti-cancer mechanism of oral cancer cells. Furthermore, JCE enhances the tumor killing effect through synergistic effect, reduces the drug dosage and reduces 5-FU resistance.