ABSTRACT
Objective To investigate the expression of microRNA-373 (miR-373) in cutaneous squamous cell carcinoma (CSCC) tissues and cells,and to explore its effects on cell invasion.Methods Real-time PCR was performed to determine the expression of miR-373 in CSCC tissues and paralesional normal skin tissues,as well as in CSCC cell lines (A431 and SCL-1) and HaCaT cells.A431 cells were divided into 4 groups:miR-373 mimic group,miR-373 inhibitor group and negative control group which were transfected with miR-373 mimic,miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.Cell invasion assay was performed to evaluate effects of miR-373 downregulation on cell invasion.Western blot analysis was conducted to assess effects of miR-373 downregulation on the protein expression of matrix metalloproteinase-2 (MMP-2) and MMP-9.Results Expression of miR-373 was significantly higher in the CSCC tissues (2.465 ± 0.218) than in the paralesional normal skin tissues (1.000 ± 0.000,P ＜ 0.05),and higher in SCL-1 cells (1.864 ± 0.178) and A431 cells (2.919 ± 0.277) than in HaCaT cells (1.000 ± 0.000,P ＜ 0.05).Most notably,miR-373 expression was also markedly higher in metastatic CSCC tissues than in non-metastatic CSCC tissues (3.323 ± 0.344 vs.1.914 ± 0.161,t =4.158,P =0.000 4).Compared with the untreated group and negative control group,the miR-373 mimic group showed significantly increased miR-373 expression and invasive ability,while the miR-373 inhibitor group showed markedly decreased miR-373 expression and invasive ability (all P ＜ 0.05).Conclusion MiR-373 downregulation can significantly suppress the invasion of A431 cells,and obviously decrease the protein expression of MMP-2 and MMP-9.
ABSTRACT
Objective To investigate the expression of microRNA-373 (miR-373) in cutaneous squamous cell carcinoma (CSCC) tissues and cells,and to explore its effects on cell invasion.Methods Real-time PCR was performed to determine the expression of miR-373 in CSCC tissues and paralesional normal skin tissues,as well as in CSCC cell lines (A431 and SCL-1) and HaCaT cells.A431 cells were divided into 4 groups:miR-373 mimic group,miR-373 inhibitor group and negative control group which were transfected with miR-373 mimic,miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.Cell invasion assay was performed to evaluate effects of miR-373 downregulation on cell invasion.Western blot analysis was conducted to assess effects of miR-373 downregulation on the protein expression of matrix metalloproteinase-2 (MMP-2) and MMP-9.Results Expression of miR-373 was significantly higher in the CSCC tissues (2.465 ± 0.218) than in the paralesional normal skin tissues (1.000 ± 0.000,P ＜ 0.05),and higher in SCL-1 cells (1.864 ± 0.178) and A431 cells (2.919 ± 0.277) than in HaCaT cells (1.000 ± 0.000,P ＜ 0.05).Most notably,miR-373 expression was also markedly higher in metastatic CSCC tissues than in non-metastatic CSCC tissues (3.323 ± 0.344 vs.1.914 ± 0.161,t =4.158,P =0.000 4).Compared with the untreated group and negative control group,the miR-373 mimic group showed significantly increased miR-373 expression and invasive ability,while the miR-373 inhibitor group showed markedly decreased miR-373 expression and invasive ability (all P ＜ 0.05).Conclusion MiR-373 downregulation can significantly suppress the invasion of A431 cells,and obviously decrease the protein expression of MMP-2 and MMP-9.