长链非编码RNA MALAT1 对胶质母细胞瘤细胞增殖和侵袭的影响及其机制
| 作者: |
1莫美绒,
2崔尖,
1蔺煦红,
1赵晓慧
1 北京中医医院顺义医院检验科，北京 101300 2 陕西省结核病医院检验科，西安 710000) |
| 通讯: |
莫美绒
Email: momeirongbj@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2018.07.002 |
摘要
目的：探讨长链非编码RNA MALAT1(long non-coding RNA MALAT1，lncRNA MALAT1)对胶 质母细胞瘤细胞增殖和侵袭的影响及其机制。方法：采用实时定量PCR(quant itative real-time polymerase chain reaction，qRT-PCR)检测胶质母细胞瘤细胞系LN-229，U87，A172，U373和 正常人脑胶质细胞系HEB中lncRNA MALAT1的表达。将LN-229细胞系分成MALAT1组与对照 组(EV组)，采用LipofectamineTM 2000分别转染pcDNA3.1-lncRNA-MALAT1质粒和阴性对照质 粒pcDNA3.1。CCK-8法测定细胞增殖能力，Transwel l试验测定侵袭能力，Western印迹法测定 基质金属蛋白酶2(matrix metalloproteinase 2，MMP-2)和pERK1/2蛋白的表达。结果：LncRNA MALAT1在胶质母细胞瘤细胞系LN-229，U87，A172，U373中的相对表达量低于正常人脑胶质 细胞系HEB。转染后0，24，48 h，MALAT1组与EV组450 nm光密度值(OD450nm)差异无统计学意 义(P>0.05)；转染后72，96 h，MALAT1组OD450nm值低于EV组，差异有统计学意义(P<0.05)。 MALAT1组侵袭细胞数为95.8±9.1，显著少于EV组(185.3±13.9)，差异有统计学意义(P<0.05)。 MALAT1组MMP-2蛋白相对表达量低于EV组(P<0.05)；MALAT1组pERK1/2蛋白相对表达量 低于EV组(P<0.05)。结论：LncRNA MALAT1在胶质母细胞瘤中下调表达，过表达lncRNA MALAT1可抑制胶质母细胞瘤细胞增殖和侵袭，其机制可能与MMP-2及pERK1/2蛋白下调表达 有关。
关键词：
胶质母细胞瘤；增殖；侵袭；长链非编码RNA MALAT1
Effect of long non-coding RNA MALAT1 on proliferation and invasion of glioblastoma cells and its mechanism
CorrespondingAuthor: MO Meirong Email: momeirongbj@126.com
DOI: 10.3978/j.issn.2095-6959.2018.07.002
Abstract
Objective: To investigate the effect and mechanism of long non-coding RNA MALAT1 (lncRNA MALAT1) on proliferation and invasion of glioblastoma cells its underlying mechanism. Methods: Real-time fluorescent quantitative PCR (qRT-PCR) was used to detect the expression level of lncRNA MALAT1 in glioblastoma cell lines (LN-229, U87, A172, U373) and normal glial cell line HEB. The LN-229 cell line was divided into the MALAT1 group and EV group, which was transfected with pcDNA3.1-lncRNA-MALAT1 and pcDNA3.1 plasmid by LipofectamineTM 2000, respectively. The proliferation and invasion ability were measured by cholecystokinin-octopeptide-8 (CCK-8) and Transwell assay, respectively. The expression level of MMP-2 and pERK1/2 protein was measured by Western blot. Results: LncRNA MALAT1 was found to be lower expressed in all 4 glioblastoma cell lines (LN-229, U87, A172, and U373), compared with normal glial cell line HEB. There was no significantly difference between MALAT1 group and EV group regarding OD450nm value after transfect for 0, 24, and 48 h (P>0.05). After transfection for 72 and 96 h. The OD450nm value of MALAT1 group was significantly lower than that in the EV group (P<0.05). The invasive cell number of the MALAT1 group was 95.8±9.1, which was significantly less than 185.3±13.9 in the EV group (P<0.05). The expression level of MMP-2 protein in the MALAT1 group was less than the EV group (P<0.05). The expression level of pERK1/2 protein in the MALAT1 group was lower than the EV group (P<0.05). Conclusion: LncRNA MALAT1 down-regulats the expression in glioblastoma cell lines. Over-expression of lncRNA MALAT1 can inhibit the proliferation and invasion of glioblastoma cell lines, which may down-regulate MMP-2 and pERK1/2 protein expression.
Keywords：
glioblastoma; proliferation; invasion; long non-coding RNA MALAT1