The Study On The Mechanism Of Sinoporphyrin Sodium-mediated Sonodynamic Therapy In Anti-proliferation Of Oral Squamous Cell Carcinoma SCC154 Cell

Background and purposeOral squamous cell carcinoma(OSCC)is the most malignant and most harmful tumor in the head and neck malignancies.Because of its special location,it will damage the oral function after clinical surgery and affect the patient’s quality of life.Sonodynamic Therapy(SDT)is a new non-invasive treatment method that combines ultrasound with a sonosensitizer to specifically kill target cells.The previous experimental results of our group showed that SDT mediated by 8μmol/L sinoporphyrin sodium(DVDMS)can inhibit the growth and proliferation of oral squamous cell carcinoma SCC154 in vivo and in vitro,and induce apoptosis of SCC154 cells in vitro.The purpose of this study was to detect DVDMS-mediated SDT damage to DNA,changes in the expression levels of DNA damage-related factors,and the expression level of mechanosensitive ion channel protein PIEZO1,and simply explored the possible mechanism of DVDMS-mediated SDT to promote cell apoptosis,and lay a theoretical foundation for the application of DVDMS-mediated SDT in the treatment of OSCC in the clinic.MethodsIn this experiment,the sonosensitizer DVDMS(8μmol/L)combined with lowintensity ultrasound(1.0MHz,duty cycle 10%,1W/cm2)was used to process SCC154 cells,and the cells were divided into four groups: control group(C),Drug alone group(D),ultrasound alone group(U),and drug combined ultrasound group(DU).Detect the distribution of DVDMS in SCC154 cells by nuclear fluorescent probe;detect the level of ROS generation in cells after different treatments by DCFH-DA probe;single-cell gel electrophoresis(comet test)was used to detect the DNA damage of SCC154 cells after different treatments;the expression levels of DNA damage factor γ-H2 A.X and DNA damage-inducing protein GADD45α in SCC154 cells after different treatments were detected by western blotting and immunofluorescence;the expression position of the mechanically sensitive ion channel protein PIEZO1 in SCC154 cells after different treatments was detected by immunofluorescence,the expression level of the mechanically sensitive ion channel protein PIEZO1 in SCC154 cells after different treatments was detected by Western blotting.ResultsThe experimental results showed that DVDMS adhered to the nuclear membrane in oral cancer SCC154 cells;after 8μmol/L DVDMS-mediated SDT treatment of SCC154 cells,compared with the control group(C),the ROS level of the combined treatment group(DU)significant increase,DNA comet tail tailing phenomenon was obvious,DNA damage factor γ-H2 A.X,DNA damage-inducing protein GADD45αexpression was increased;mechanosensitive ion channel protein PIEZO1 was mainly expressed on the cell membrane,combined treatment group(DU)increased expression of mechanosensitive ion channel protein PIEZO1ConclusionsDVDMS-mediated SDT can damage the DNA of oral squamous cell carcinoma SCC154 cells and increase the expression of cell DNA damage repair related protein GADD45α;DVDMS-mediated SDT can increase the expression of mechanosensitive ion channel protein PIEZO1.