The Inhibition Effect Of Cold Atmospheric Plasma-Activated Media In Cutaneous Squamous Carcinoma Cells

Objective Cutaneous squamous cell carcinoma(SCC)is a common nonmelanoma skin cancer t,usual suffered old people.According to the latest data,the increasing incidence and mortality of skin squamous cell carcinoma make the treatment of this disease become the focus of clinical dermatology.The main clinical treatment methods include operation,chemotherapy and radiotherapy alone or in combination.Although the treatment effect of most patients is satisfactory,especially the young patients.However,the prognosis of patients with invasive cutaneous squamous cell carcinoma is poor,especially for the elderly for most patients cannot tolerate the three treatments mentioned above and miss the best treatment time.CAP activated media was produced by the cold plasma(CAP)treatment.In recent years,many studies have confirmed that it has significant inhibitory effect on various tumor cells in vivo and vitro,and has little effect on normal tissue.In this study,PBS and DMEM was treated by the CAP device for different time and the effect of the activated media on human skin squamous cell carcinoma cells and the related mechanism were observed.Method This study included two parts:the study of cell viability and its related mechanism.Firstly,CAP device was used to treat PBS and DMEM respectively to obtain CAP-PBS and CAP-DMEM on Os,60s,120s,180s,240s,300s and 360s.A431 cells and HaCat cells were immediately treated with CAP media,and the effect of CAP media on the proliferation of skin squamous cell carcinoma cells and normal skin keratinocytes was determined by MTT assay.At the same time,the plasma activation solution will be prepared.In this study,Annexin V and PI assay were used to detect apoptosis,and Western blot was used to detect the expression of PARP and caspase-7.CM-H2DCFDA probe was used to detect the intracellular reactive oxygen(ROS)level in A431 cells treated by CAP media.Results MTT results showed that CAP-PBS and CAP-DMEM significantly inhibited the proliferation of A431 cells in a dose-dependent and time-dependent manner,but the inhibition of HaCat cells was weak.At the same time,the stored CAP media still showed significant inhibitory effect.Annexin-V and PI and Western blot results showed that the inhibitory effect of CAP on A431 cells was achieved by inducing apoptosis.CM-H2DCFDA probe was used to measure intracellular ROS level after CAP media treatment,and the results showed that the intracellular ROS level of A431 increased after CAP-PBS treatment,and the intracellular ROS level increased significantly with the increase exposure time of CAP treatment time,but CAP-DMEM had little effect on the intracellular ROS level of A431.Conclusion CAP media has obvious inhibitory effect on 431 cells,and this inhibitory effect is produced by apoptosis.The increase of intracellular ROS induced by CAP may be one of the factors induce apoptosis.