Role of the RECK Gene in the Inhibition of Cutaneous Malignant Melanoma Proliferation by Vemurafenib
Objective:To explore the role of the reversion inducing cysteine rich protein with Kazal motifs(RECK)gene in inhibit-ing of cutaneous malignant melanoma(CMM)proliferation by vemurafenib.Methods:The expression of RECK gene in CMM tissue and adjacent tissue was verified through bioinformatics analysis,and divided the sample into low expression and high expression groups based on the expression of RECK gene in CMM tissue.The differences between the RECK gene low expression group and the high expression group were analysed by Kaplan Meier survival curve.Human malignant melanoma A375 cells were used to construct a nude mouse transplanted tumor model.When the tumor volume reached about 100 mm3,the tumor-bearing mice were divided into vemu-rafenib group(75 mg/kg/day gavage)and the control group(equal amount of solvent gavage),with 6 mice in each group.After treatment,CMM tissue was collected,and real-time fluorescence quantitative PCR was used to detect and compare the mRNA expression levels of RECK gene between the verbenib group and the control group.Western blot and immunohistochemical staining were used to detect and compare the differences in RECK protein expression levels between the remofenib group and the control group.Results:Bioinformatics analysis showed that the expression level of RECK gene in CMM tissues was significantly lower than that in adjacent tissues,with statis-tical significance(P＜0.001).Kaplan Meier survival curve analysis showed that the level of RECK gene expression was correlated with the prognosis of CMM patients,the survival time of the RECK gene low expression group was shorter than that of the high expression group(P=0.007).After 16 consecutive days of administration,the relative tumor proliferation rate and tumor inhibition rate of the vemurafenib group were 37.60％and 63.45％,respectively,and successfully constructed a CMM nude mouse tumor inhibition model for transplanta-tion.The relative amounts of RECK mRNA in the vemurafenib group and the control group were 1.02±0.13 and 0.99±0.15,respectively,with no statistically significant difference(P＞0.05);The expression of RECK protein in the vemurafenib group and the control group was 0.19±0.01 and 0.18±0.01,respectively,with no statistically significant difference(P＞0.05);Immunohistochemical staining showed that the RECK gene was negatively expressed in both the vemurafenib group and the control group.Conclusion:The RECK gene is correlated with the development and prognosis of CMM,but there is no significant change in RECK during the inhibition of CMM proliferation by veimofenib.
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