The Research On The Effect Of Surfactin On Proliferation、Apoptosis And Migration Of Tongue Squamous Cell Carcinoma Tca8113

Objective: Preliminarily investigate the effect of Bacillus subtilis fermentation products Surfactin on the occurrence and progress of tongue squamous cell carcinoma Tca8113 cell line.Methods:To detect the status of cell growth,When the tongue squamous cancer cells grow to about 80%, added 20 μg/m L、40 μg/m L、60 μg/m L、80 μg/m L、100 μg/m L concentration of drugs,the proliferation effect on tongue squamous cell carcinoma Tca8113 cell was checked by Sulforhodamine B assay when the cells was cultured 24 h and 48h;Clonogenic assay was used to detected the effect of Surfactin on the cloned numbers of tongue squamous cell carcinoma;Morphological changes of tongue squamous cell carcinoma Tca8113 cell was observed by Hochest/PI staining fluorescence microscope;Cell early apoptosis was measured by flow cytometry; The change of the migration distance of tongue squamous cell carcinoma Tca8113 cell was observed by wound healing assay,Transwell migration assay was used to detected the influence of the migration of surfactin for tongue squamous cancer cells;The expression of matrix metalloproteinase MMPs was verified by Western blot assay.Result:1. The SRB assay display that surfactin can effectively inhabit the proliferation of tongue squamous cell carcinoma Tca8113 cell with dose-dependent manner,and the IC50 were74μg/m L and 53μg/m L for 24 and 48 hours respectively;2. Clony formation assay results show that Surfactin can inhibit the tongue squamou s cancer cell clone formation, when the concentration of Surfactin were 25μg/m L、50μg/m L、100μg/m L respectively, cloning aggregation rate of 28.3%±0.42%、26.5%±0.71%、20.7%±0.42%, surfactin can strongly inhibit the number of cell clones.3. Hochest/PI double fluorescent the results showed that the cells highlighting,nuclear pyknosis.The flow cytomery detected that when Surfactin concentration of 25μg/m L、50 μg/m L、100μg/m L respectively,the early apoptosis rate were 11.4%、43.45、72.74%.Compared with control group,drug intervention group cell apoptosis rate in creased significantly.4. Healing wound assay observe that Surfactin can significantly inhibit the healing o fcell scratches;5. Transwell Migration assay results show that the Migration cells were 429.33±28.92、234.44±7.02、141.67±29.40、52.000±17.69 when the Tongue squamous cell carcinoma cell Tca8113 were exposed to the drug concentration of 0μg/m L、25μg/m L、50μg/m L、100μg/m L respectively,With the increase of drug concentration,Cell migration distance was decreasing.6. Western blotting results show that Surfactin can inhibit Tca8113 cell migration by down-regulating the expression of MMPs.Conclusion:Surfactin may as a potential treatment for recurrent and metastatic tongue cancer patients...