Study On The Inhibition Of CAL27 Cell Migration And Epithelial Stromal Transformation By Cold Atmosphere Plasma Through JNK/STAT3/MMP9 Signaling Pathway

Objective : Atmospheric pressure low temperature plasma technology has many advantages,such as high efficiency,low energy consumption,no pollution,close to normal temperature environment and no need of complex vacuum system.It is currently used in many fields,such as disinfection and sterilization,material surface modification,film deposition,environmental purification and food safety.In recent years,there has been a huge wave in the field of medicine.The atmospheric pressure plasma(CAP)is an electrically neutral ionized gas composed of positive and negative charged particles and a variety of neutral particles.Cap is also involved in the transformation of some malignant tumors,which brings a new dawn for clinical treatment of malignant tumors.Tongue cancer is a kind of head and neck cancer which is prone to lymph node metastasis in two thirds of men.The purpose of this paper is to study the effects of low temperature plasma on the proliferation,migration and invasion of tongue cancer cells,and to provide new technical support and theoretical basis for clinical treatment of tongue cancer.Methods:CAP was used to treat cal27 cells for 0s,15 s,30s and 60 s respectively,and the morphology of cal27 cells was observed under inverted microscope at different stimulation time;DCFH-DA probe was loaded under different treatment conditions,and ROS content in cells was observed under fluorescence microscope;CCK-8 technology was used to calculate cell proliferation capacity;cell scratch test and Transwell method were used to detect Western blot was used to detect the expression of migration related proteins in tongue cancer cells.The data were calculated by one-way ANOVA and analyzed by SPSS.Results:1.It was found that under the inverted microscope,cap could make the cells smaller and rounder.When the treatment time reached 60 s,the attachment ability of some adherent cells to the basement decreased and they floated in a mass.2.The results of fluorescence microscopy showed that the content of ROS in tongue cancer cells increased with the increase of stimulation time,which indicated that cap stimulationcould increase the content of ROS in a certain period of time.3.The results of CCK-8showed that cap stimulated the proliferation of tongue cancer cells significantly(P <0.001),and the effect of this change was related to the stimulation time.4.The results of cell scratch technique and Transwell experiment indicate that the time of cap stimulation can inhibit the migration of tongue cancer cells to a certain extent.(P<0.05)。 5.The results of Western blot showed that the content of E-cadherin increased,the content of STAT3,JNK,N-cadherin and MMP9 decreased.The results showed that cap could inhibit the migration of tongue cancer cells(P < 0.05).When the stimulation dose was60 s,the migration related protein(EMT)changed significantly(P < 0.05).Conclusion:Atmospheric pressure plasma cap can improve the ROS content of tongue cancer cells,and the migration of tongue cancer cells is inhibited by atmospheric pressure plasma to some extent.At the same time,it can inhibit the proliferation of tumor cells to a certain extent.