S1pr1沉默对弥漫大B细胞淋巴瘤OCI-LY1细胞增殖和迁移的影响

Effects of S1pr1 silencing on the cell proliferation and cell migration of diffuse large B-cell lymphoma OCI-LY1 cells

目的探讨S1pr1沉默对弥漫大B细胞淋巴瘤(DLBCL)OCI-LY1细胞增殖和迁移的影响。方法收集确诊为DLBCL的患者和健康体检者各10例,测定外周血S1pr1 mRNA的含量,用S1pr1特异性siRNA和阴性对照(negative control,NC)转染到OCI-LY1细胞,用RT-PCR和Western Blot法检测OCI-LY1细胞中S1pr1的表达,以鉴定S1pr1特异性siRNA沉默效果。检测OCI-LY1细胞增殖率和细胞迁移能力,同时检测STAT3和p-STAT3蛋白表达水平。结果DLBCL患者外周血单个核细胞中S1pr1 mRNA相对中位表达水平高于健康者(P<0.05);给予S1pr1特异性siRNA干预后,S1pr1沉默组OCI-LY1细胞中S1pr1 mRNA和蛋白表达较空白对照组和阴性对照组显著降低(P<0.05);S1pr1沉默组OCI-LY1细胞增殖率和迁移细胞数较空白对照组和阴性对照组显著降低(P<0.05);S1pr1沉默组OCI-LY1细胞STAT3和p-STAT3蛋白表达较空白对照组和阴性对照组显著降低,差异有统计学意义(P<0.05)。结论S1pr1在DLBCL患者外周血中高表达,对S1pr1基因沉默后,OCI-LY1细胞增殖率和迁移细胞数显著降低,特异性抑制了STAT3的活性。

Objective To investigate the effects of S1pr1 silencing on the cell proliferation and cell migration of diffuse large B-cell lymphoma(DLBCL)OCI-LY1 cells.Methods Ten patients with DLBCL and 10 healthy subjects were enrolled in the study.The mRNA levels of S1pr1 in peripheral blood were detected.S1pr1 specific siRNA and negative control(NC)were transfected into OCI-LY1 cells.The expression levels of S1pr1 in OCI-LY1 cells were detected by RT-PCR and Western Blot to identify the silencing effects of S1pr1 specific siRNA.Moreover the proliferation and migration abilities of OCI-LY1 cells were detected,and the expression levels of STAT3 and p-STAT3 protein were also detected.Results The relative median expression levels of S1pr1 mRNA in peripheral blood mononuclear cells of patients with DLBCL were significantly higher than those in healthy subjects(P<0.05).After the intervention of s1pr1-specific siRNA,the mRNA and protein expression levels of S1pr1 in OCI-LY1 cells in S1pr1 silencing group were significantly lower than those in blank control group and the negative control group(P<0.05).The proliferation rate and migration number of OCI-LY1 cells in S1pr1 silencing group were significantly lower than those in blank control group and negative control group(P<0.05).In addition the expression levels of STAT3 and p-STAT3 protein in OCI-LY1 cells in S1pr1 silencing group were significantly lower than those in blank control group and negative control group(P<0.05).Conclusion S1pr1 is highly expressed in peripheral blood of patients with DLBCL.After the silencing of S1pr1 gene,the cell proliferation rate and migration number of OCI-LY1 cells are significantly decreased,which inhibits specifically the activity of STAT3.