Gene Of Human Signal Of Semaphorin-3f Transfected With Tongue Squamous Cell Carcinoma Cell Proliferation

The human semaphorins were originally characterized as constituents of the complex regulatory system responsible for the guidance of growing axons to their targets during the development of the central nervous system.The semaphorins have been implicated in diverse developmental processes,such as axon guidance during nervous system development and regulation of vascular endothelial cell development and migration.The semaphorin-3F(SEMA-3F)is class 3 of the large family of genes,which is a type of secretion type proteinum to be representative and researched transparently.They reside in multiple cells and tissues. Recently,the SEMA-3F function additionally as antioncogene,the gene express down regulation or deletion in multiple tumour cells,such as small cell lung cancer,malignant melanoma and squamous cell carcinoma of the head and neck.SEMA-3F may play a potent regulatory role in tumorigenesis and in the process of tumor formation.The event showed that the SEMA-3F gene may express down regulation similarity in squamous cell carcinoma of tongue and inhibit the proliferation of Tca8113 cells.In this experiment,after constructed a full length SEMA-3F expression vector,Tca8113 cells were transfected by Lipofectamine 2000 regent,a kind of positive ion lipoplast transfection regent.To screen the Tca8113 cells with SEMA-3F and detect the expression of SEMA-3F gene.We investigated the proliferation of Tca8113 cells by methyl thiazolyl tetrazolium(MTT)and flow cytometer (FCM)after transfection.Methods:Tca8113 cells were equally divided into four groups: transfected cell groups,empty vector groups,untransfected cell groups and control groups(exclusively transfection regent).After construction of a full-length human SEMA-3F expression vector,Tca8113 cells were transfected with pEGFP-N1-SEMA-3F by Lipofectamine 2000 regent. The cells were screened by G418(400μg/mL)48 hours later,lasted 4 weeks and supersede culture fluid every 3 to 5 days.The expression of SEMA-3F gene in the cells was detected by Western Blotting and RT-PCR.The survival rates of different groups were assayed by MTT (methyl thiazolyl tetrazolium)enzymatic labeling technique.Cell cycles were assayed by flow cytometer(FCM).Results:14 days after the selection,G418 cell clones containing SEMA-3F was identified.The gene was transfected into Tca8113 cells. The results of Western Blotting and RT-PCR revealed a steady high expression of the SEMA-3F in the transfected cells.The survival rate of the transfected cell groups was decreased significantly than other groups. The cell cycle percentage of G₁ stage was to degrade and S stage was to increase.Conclusion:The expression of semaphorin-3F gene is extremely weak in Tca8113 tongue carcinoma cells.The cells can steadily express exogene SEMA-3F by Lipofectamine 2000 regent transfection. SEMA-3F gene transfection may inhibit the proliferation of Tca8113 cells.