Expression And Significance Of Hsa-miR-100-5P In Skin Squamous Cell Carcinoma And Arsenite-induced HaCaT Malignant Epithelial Cells

Objective:Analysis of hsa-mi R-100-5P expression in cutaneous squamous cell carcinoma cells(c SCC cells)and arsenic-induced Ha Ca T malignant phenotype cells(AS-TM cells)to verify the possibility of AS-TM cells becoming arsenic-induced skin cancer models,to explore the common molecular basis between arsenic-induced Ha Ca T cell malignancy and skin squamous cell carcinoma,and to provide a new target for the diagnosis and treatment of cutaneous squamous cell carcinoma.Methods:1.Preparation of AS-TM cells: Ha Ca T cells were cultured in a 100 n M arsenic trioxide culture medium for 180 days,and cells were collected and then prepared into AS-TM cells.The soft agar colony formation assay was used to calculate the colony formation rate.2.Detection of hsa-mi R-100-5P expression: Normal Ha Ca T cells,AS-TM cells,and c SCC cells were subcultured at the same time.Three types of cells in the logarithmic growth phase were selected and divided into three groups.Real-time quantitative polymerase chain reaction(RT-PCR)was used to detect the relative expression of hsa-mi R-100-5P in human immortalized keratinocytes(Ha Ca T cells),cutaneous squamous cell carcinoma cells(c SCC cells),and arsenic-induced Ha Ca T malignant phenotype cells(AS-TM cells).Results:1.The Ha Ca T cells cultured in arsenic-containing medium for 180 days showed significant changes in morphology.Under the microscope,the volume of the cells became smaller,the cell morphology varied,and multinucleated giant cells frequently appeared.Ha Ca T cells in the arsenic-free control group were still epithelioid cells.The colony formation rate of AS-TM cells group in soft agar colony formation assay was significantly higher than that of Ha Ca T cell group,and the monoclonal formation rates of Ha Ca T cell group and AS-TM cell group were 0.12� and 0.52�,respectively.There was a significant difference between the two groups.(P<0.05).2.RT-PCR results showed that there was a significant difference between the expression levels of hsa-mi R-100-5P in the three groups of cells by one-way ANOVA(F=44.866,P=0.000*).Multiple comparison using the LSD-t method showed that hsa-mi R-100-5P was significantly up-regulated in c SCC cells compared to Ha Ca T cells(LSD-t = 8.344,P = 0.000*),the relative expression levels of which in c SCC cells and in Ha Ca T cells were 2.445�0.261 and 1.000�0.000,respectively.Compared with Ha Ca T cells,the relative expression level of hsa-mi R-100-5P in AS-TM cells was also significantly up-regulated(LSD-t = 3.893,P = 0.008*),the relative expression levels of which in AS-TM cells and in Ha Ca T cells were 1.636�0.192 and 1.000�0.000,respectively.Furthermore,the expression of hsa-mi R-100-5P was significantly up-regulated in c SCC cells compared with AS-TM cells(LSD-t = 4.452,P = 0.004*).The results demonstrated that the relative expression of hsa-mi R-100-5P was up-regulated in c SCC cells and AS-TM cells compared with normal Ha Ca T cells.Conclusions:1.Hsa-mi R-100-5P was significantly up-regulated in arsenic-induced Ha Ca T cell malignant phenotypic cells and cutaneous squamous cell carcinoma cells,indicating that there was a similar expression of mi RNA between arsenic-induced Ha Ca T cell malignant phenotypic cells and cutaneous squamous cell carcinoma cells,which confirmed that arsenic-induced Ha Ca T cell malignant phenotypic cells were reliable as arsenic-induced skin cancer cell models.2.Hsa-mi R-100-5P may play a role in the malignant transformation of arsenic-induced Ha Ca T cells and the development of cutaneous squamous cell carcinoma,and may be a potential target for the diagnosis and treatment of cutaneous squamous cell carcinoma.