Biological Effects Of Gene X Overexpressing On OLK Cell

Oral squanmous cell carcinoma is the most common tumor in oral maxillofacial with high incidence,high recurrence rate,poor prognosis and a high rate of lymph node metastasis.Oral squanmous cell carcinoma continues to difficult to treat and conventional treat strategies result in a plethora of side effect that affect normal physiological functions including speech,swallowing and physical appearance.Moreover,aggressive therapy is often difficult to avoid the occurrence of comorbidities in oral squanmous cell carcinoma patients.The etiology of oral squanmous cell carcinoma is various and a small amount is derived from oral leukoplakia.Oral leukoplakia is the most common oral precancerous lesion which is resulted in smoking and alcohol consumption mainly.Oral leukoplakia is diagnosed mainly by histopathologic examination clinically and lack of other effective way to early detect the development and progression of the tumor.Therefore,looking for effective biomarkers in order to early detect malignant change is an important means to reduce the incidence of oral squanmous cell carcinoma.Objective: This research is aimed to investigate the biological functions of Gene X on proliferation and migration on DOK cell by overexpressing Gene X.Methods: Overexpression plasmid GV141-gene X vector was constructed and transfected into DOK cell with LipofectamineTM 2000 reagent.The m RNA and protein expression levels were detected by Real time PCR and Western blot,respectively.The effect of gene X on proliferation and migration were analyzed by Proliferation assay and Transwell assay,respectively.The expression of cell cycle protein Cyclin B1 was detected by Western blot too.And the Geneticin G418 was used to screen the stable expression cell line,then the target protein was indentified by Western blot.Result: More m RNA and protein of Gene X were expressed in OSCC-BD cell than DOK cell,The recombinant plasmid vector was successfully constructed,both m RNA and protein of gene X were successfully expressed in DOK cell.The proliferation of DOK cell with reconstituted gene X was significantly enhanced as compared with the control cell.The migration of DOK cell with reconstituted gene X was significantly increased compared with the control cell(P<0.05).The expression the cell cycle protein cyclin B1 was increased.The recombination protein was expressed successfully,while the control cell did not express.Conclusion: Gene X can enhance the ability of proliferation and migration of DOK cell,these results imply Gene X may play an important role in the development and metastasis of oral squamous cell carcinoma.The increasing expression of Cyclin B1 caused by Gene X overexpressing may imply the Gene X relates to the cell cycle regulation.And the stable expression cell line was successfully established.