下调人类白细胞抗原复合体18靶向微小RNA-146b-5p对胶质瘤SHG44细胞迁移和侵袭的影响

Effects of down-regulating human leucocyte antigen complex group 18 targeting microRNA-146b-5p on migration and invasion of glioma SHG44 cells

目的研究下调长链非编码RNA (lncRNA)人类白细胞抗原复合体18(HCG18)靶向微小RNA-146b-5p(miR-146b-5p)对胶质瘤SHG44细胞迁移和侵袭的影响。方法以实时定量-PCR法检测胶质瘤SHG44、CHG-5和BT325细胞与正常胶质HEB细胞中HCG18基因表达,用SHG44细胞做后续实验。将SHG44细胞分成7组:空白对照组、第1转染组(转染shRNA control)、第2转染组(转染HCG18 shRNA)、第3转染组(共转染HCG18 shRNA、inhibitor control)、第4转染组(共转染HCG18 shRNA、miR-146b-5p inhibitor),第5转染组(转染mimics control)和第6转染组(转染miR-146b-5p mimics),均为100 pmol。Transwell小室检测细胞侵袭和迁移能力变化。荧光素酶报告系统鉴定HCG18和miR-146b-5p靶向关系。结果 HCG18基因在胶质瘤SHG44、CHG-5、BT325细胞中表达水平分别为3.68±0.24,2.73±0.22和1.95±0.13,均高于正常胶质HEB细胞的1.00±0.11,差异均有统计学意义(均P<0.05),且胶质瘤SHG44细胞中HCG18表达水平最高。空白对照组、第1转染组、第2转染组、第3转染组和第4转染组细胞侵袭数目分别为156.32±12.58,155.80±13.97,99.76±8.52,100.20±9.63和146.16±10.58;迁移数目分别为196.74±15.52,199.86±16.39,148.13±13.28,150.89±10.76和181.72±12.39。第2转染组的上述指标与第1转染组相比,或第4转染组的上述指标与第3转染组相比,差异均有统计学意义(均P<0.05)。经wt处理的第6转染组和第5转染组细胞荧光素活性分别为1.00±0.11和0.41±0.04,第6转染组与第5转染组相比,差异有统计学意义(P<0.05)。结论下调长链非编码RNA HCG18靶向miR-146b-5p可抑制胶质瘤SHG44细胞侵袭和迁移。

Objective To study the effect of down-regulating long non-coding RNA(lncRNA)human leucocyte antigen complex group 18(HCG18)targeting microRNA-146 b-5 p(miR-146 b-5 p)on the migration and invasion of glioma SHG44 cells.Methods Real time quantitative-PCR was used to detect the changes of HCG18 gene expression in glioma SHG44,CHG-5,BT325 cells and normal glial HEB cells.SHG44 cells were used for follow-up experiments.The SHG44 cells were divided into 7 groups:blank control group,transfection-1 group(transfected with shRNA control),transfection-2 group(transfected with HCG18 shRNA),and transfection-3 group(co-transfected with HCG18 shRNA,inhibitor control),transfection-4 group(co-transfection of HCG18 shRNA,miR-146 b-5 p inhibitor),transfection-5 group(transfected with mimics control),and transfection-6 group(transfected with miR-146 b-5 p mimics),with 100 pmol in each group.Transwell cells were used to detect changes in cell invasion and migration ability.The luciferase reporter system identified HCG18 and miR-146 b-5 p targeting relationships.Results The expression of HCG18 gene in glioma SHG44,CHG-5,and BT325 cells were respectively 3.68±0.24,2.73±0.22,1.95±0.13,higher than that of normal glioma HEB cells(1.00±0.11)with significant difference(all P<0.05),and the expression of HCG18 was highest in SHG44 cells.The number of cell invasions in the blank control group,transfection-1 group,transfection-2 group,transfection-3 group,and transfection-4 group were 156.32±12.58,155.80±13.97,99.76±8.52,100.20±9.63,146.16±10.58,respectively;the number of migrations were 196.74±15.52,199.86±16.39,148.13±13.28,150.89±10.76,181.72±12.39,respectively.Compared between transfection-2 group and transfection-1 group,or comparison between transfection-4 group and transfection-3 group,the difference of the above indicators were significant(all P<0.05).The cytofluorescein activity of the transfection-5 group and the transfection-6 group treated with wt were 1.00±0.11,0.41±0.04,compared between transfection-6 group and transfection-5 group,the difference was statistically significant(P<0.05).Conclusion Down-regulating HCG18 targets miR-146 b-5 p can inhibit glioma SHG44 cell invasion and migration.