Objective:To investigate the effect of miR-125b targeting Sema4C on STAT3 signaling pathway on invasion and metastasis of non-Hodgkin′s lymphoma.Methods: Expression of miR-125b in non-Hodgkin′s lymphoma tissues and cell lines was detected by QT-PCR.Expression of Sema4C in normal lymphocyte tissues and non-Hodgkin′s lymphoma tissues was detected by immunohistochemistry.Dual luciferase effect of miR-125b on the transcriptional activity of Sema4C was examined by the reporter gene system.Transwell invasion assay was used to detect the expression of miR-125b in the non-Hodgkin′s lymphoma cell line NK-92.Scratch test Western blot was used to detect the expression of Sema4C.Western blot was used to detect the protein expression of STAT3 signal pathway after silencing Sema4C.The protein expression of Sema4C was detected by Western blot.Results: Expression of miR-125b was significantly lower in non-Hodgkin′s lymphoma tissues than that in normal tissues[(0.48±0.05)% vs (1.59±0.38)%,P<0.05].Sema4C was highly expressed in non-Hodgkin′s lymphoma[(326.25±7.75)% vs (58.75±5.76)%].After overexpression of miR-125b,non-Hodgkin′s lymphoma cells expression level of Sema4C was down-regulated after silencing Sema4C[(326.25±7.75)% vs (58.75±5.76)%],and the expression of JAK and STAT3 protein were down-regulated after miR-125b overexpression[(85.26±6.94)% vs (12.61±4.32)%,P<0.05].The dual luciferase reporter gene system showed that miR-125b could directly regulate the transcriptional activity of Sema4C.Conclusion: miR-125b can regulate the invasion and migration of non-Hodgkin′s lymphoma cells by targets the expression of Sema4C.%目的:研究miR-125b靶向Sema4C调控STAT3信号通路影响非霍奇金淋巴瘤的侵袭迁移.方法:运用QT-PCR法检测miR-125b在非霍奇金淋巴瘤组织和细胞株中的表达;运用免疫组化法检测Sema4C在正常淋巴组织和非霍奇金淋巴瘤组织中的表达;用双荧光素酶报告基因系统检测miR-125b对Sema4C转录活性的影响;用Transwell小室侵袭实验和划痕实验检测miR-125b的表达对NK-92细胞侵袭能力的影响;用 Western blot法检测过表达miR-125b后NK-92细胞Sema4C的表达变化;Western blot法检测沉默Sema4C后NK-92细胞STAT3信号通路的蛋白表达水平变化.结果:与正常淋巴组织比较,miR-125b在非霍奇金淋巴瘤组织中表达明显降低[(0.48±0.05)% vs (1.59±0.38)%,P<0.05],Sema4C在非霍奇金淋巴瘤中表达较高[(1.36±0.25)% vs (0.34±0.08)%,P<0.05];过表达miR-125b后,NK-92细胞的侵袭和转移能力明显降低[(326.25±7.75)% vs (58.75±5.76)%],Sema4C的表达水平下调[(84.26±6.94)% vs (15.61±4.68)%,P<0.05].沉默Sema4C后,NK-92细胞JAK和STAT3蛋白表达下调[(85.26±6.94)% vs (12.61±4.32)%,P<0.05];双荧光素酶报告基因系统检测结果显示,miR-125b可以直接调控Sema4C的转录活性.结论:在非霍奇金淋巴瘤组织和细胞株中,miR-125b可靶向Sema4C的表达,从而调控非霍奇金淋巴瘤细胞的侵袭和迁移能力.
展开▼
