Effects Of Qilan Preparation On Cell Proliferation And Apoptosis In Premalignant Human Oral Epithelial Cells

Objective: Different concentrations of Chinese herbal medicine Qilan Preparation were used to act on human oral keratinocytes(HOK)、dysplastic oral keratinocyte cells(DOK)、 human oral squamous cell carcinoma cells(Tca8113), investigating whether Qilan Preparation has selective inhibition on three different cancer progression state cell lines and studying the proliferation and apoptosis mechanism of DOK induced by Qilan Preparation for clinical applications. Methods:: 1. Culturing HOK、DOK、Tca8113, three different cancer progression state cell lines. 2. The growth inhibition rate of HOK、DOK 、Tca8113 was assayed by CCK-8. 3. The cell cycle and apoptosis in DOK cells induced by different concentrations of Qilan Preparation was analyzed by flow cytometry. 4. The expression of Bax protein and Bcl-2 protein、Cyclin D1 protein were analyzed by Western blot. Results: 1. Staining for keratin was observed in HOK, HOK、DOK、Tca8113 have different growth characteristics. 2. CCK-8 assay showed Qilan preparation had cell proliferation inhibition effect on HOK、 DOK、 Tca8113 in a dose-dependent manner. the inhibition strength was Tca8113>DOK>HOK. 3. The results of flow cytometry showed that the apoptosis rate increased with the increasing of Qilan Preparation concentration. Qilan Preparation induced G0/G1 arrest in DOK cell. 4. The results of WB showed that the expression of Cyclin D1 protein、Bcl-2 protein was significantly down-regulated after treated with Qilan Preparation for 24 hours,while the expression of Bax protein and Bax/Bcl-2 was up-regulated. Conclusion: 1. Successful culture HOK、DOK、 Tca8113。 Three cell lines have different growth characteristics and different cancer progression state. 2. Qilan Preparation could significantly inhibit proliferation of HOK、DOK、Tca8113. 3. Qilan Preparation has low toxicity and selective inhibition on three kinds ofcells, the more maligmant,the more effective. 4. Qilan Preparation could promote DOK cell apoptosis. 5. Qilan Preparation inhibits DOK cell proliferation might through G0/G1 arrestby alteration in EGFR、P27 and Cyclin D1. 6. Down-regulation of Bcl-2 protein expression and up-regulation of p53-dependent Bax protein expression result in DOK cells apoptosis,which might be one of the main mechanisms for apoptosis of DOK cells.