| Background/Objectives: Skin squamous cell carcinoma(SCC)is the second most common malignant skin cancer in the world after basal cell carcinoma(BCC).In recent years,it has an obvious upward trend worldwide.The pathogenesis of this disease is not clear.Ultraviolet irradiation,burn(trauma),precancerous dermatosis,human papillomavirus infection and so on may be related to the occurrence of this disease.At present,the common treatment methods include surgical excision,laser,cryotherapy and so on.It is easy to form large scars after operation,and the recurrence rate of laser and cryotherapy is high.In addition,there is no satisfactory treatment for patients with large area,metastasis or special site.Itraconazole is a commonly used antifungal drug.In recent years,studies have shown that itraconazole has a good inhibitory effect on hemangioma,non-small cell lung cancer,basal cell carcinoma,melanoma,and cervical squamous cell carcinoma cells.Therefore,we speculate that itraconazole may also inhibit SCC.The purpose of this study is to explore the effect of itraconazole on SCL-1 cells and provide new ideas and basis for the treatment of SCC.Methods: 1.The morphological changes of SCL-1 cells before and after itraconazole treatment were observed under inverted microscope.2.CCK8 assay was used to detect the inhibitory effect of itraconazole on SCL-1 cells.The suspension of SCL-1 cells was cultured in 6-well plate.After cell adherence(about 8 hours),itraconazole was added into each hole,and the concentration of itraconazole was 0μM,1μM,2μM,4μM,8μM,respectively.CCK8 was detected at 0,24,48,72 and 96 hours.OD value was measured and cell inhibition level was calculated.3.Flow cytometry was used to detect cell apoptosis.Cells in each group were collected at 12,24 and 36 hours,respectively,and the apoptosis level was detected by computer.Results: 1.The cells in the control group were polygonal and plump in shape.The organelles and large nuclei could be seen in the cells.The cells grew adherently and connected tightly.In the experimental group,some cells became round,some were long spindle-shaped,the volume was reduced and bright,the cell membrane was ruptured,organelles and nuclei flowed out,the number of cells was significantly reduced,and a large number of dead cell debris was suspended in the culture medium.In addition,a special "chrysanthemum-like" structure was observed outflowing from nuclears under the microscope.2.The results of CCK8 experiment showed that the OD value of the control group increased with time and the growth curve was similar to "s".The OD value of the experimental group increased first and then decreased,and the growth curve was close to inverted "v".At 72 hours,there was no significant difference in OD value and inhibition rate between 2 u M and 8 u M groups(p > 0.05).At 96 hours,the OD value of 2 u M and 4 u M groups were the lowest,the inhibition rate was the highest.There were no significant difference in OD value and inhibition rate between the two groups.(p > 0.05).3.When itraconazole was treated for 12,24 and 36 hours,no obvious apoptotic peak was found,but the cell necrosis rate increased with the increase of time and drug concentration.Conclusion: 1.Itraconazole has an inhibitory effect on SCL-1 cells.In a certain range,the inhibition increased with time.2.The inhibitory effect of itraconazole on SCL-1 cells was not positively correlated with the concentration of itraconazole.3.Itraconazole has no significant effect on apoptosis of SCL-1 cells,but can promote cell necrosis.The mechanism of inhibiting SCL-1 cell proliferation and promoting SCL-1 cell necrosis remains to be further studied.4.Unlike previous cell death modes,the outflow of chrysanthemum-like structure from nuclears is the characteristic expression of dead cells in this experiment,which may be a new mode of cell death. |
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