SNHG1靶向调控miR-145对神经母细胞瘤细胞增殖、侵袭和迁移能力的影响

Effects of lncRNA SNHG1 targeting miR-145 on proliferation, invasion and migration of neuroblastoma cells

目的:探讨核仁小分子RNA宿主基因1(SNHG1)对神经母细胞瘤SK-N-SH细胞增殖、侵袭、迁移的影响及分子机制。方法:用Lipofectamine2000将SNHG1 shRNA(sh-SNHG1组)或阴性对照shRNA(sh-NC组)分别转染入SK-N-SH细胞中,以未转染细胞为空白对照。转染24 h后,qRT-PCR法检测细胞中SNHG1的表达,CCK-8测定细胞增殖能力,Transwell小室实验检测细胞侵袭和迁移能力,Western blot法测定细胞中MMP-9、MMP-2蛋白的表达。利用在线靶基因预测软件Starbase预测SNHG1与miR-145的结合位点,双荧光素酶报告实验鉴定两者的靶向关系。将SNHG1 shRNA、miR-145抑制物共转染到SK-N-SH细胞中(sh-SNHG1+anti-miR-145组),将SNHG1 shRNA、抑制物阴性对照共转染到SK-N-SH细胞中(sh-SNHG1+anti-NC),测定细胞增殖、侵袭、迁移能力,以及细胞中miR-145、MMP-9和MMP-2蛋白的表达。结果:与空白对照、sh-NC组比较,sh-SNHG1组细胞增殖、侵袭和迁移能力,细胞中SNHG1、MMP-2和MMP-9蛋白表达水平下降(P<0.05)。双荧光素酶报告实验结果证实SNHG1靶向负调控SK-N-SH细胞中miR-145的表达。与sh-SNHG1+anti-NC组比较,sh-SNHG1+anti-miR-145组细胞miR-145表达水平降低(P<0.05),MMP-2和MMP-9表达增强,细胞增殖、侵袭和迁移能力增强(P<0.05)。结论:SNHG1可通过靶向负调控miR-145的表达,促进神经母细胞瘤细胞的增殖、侵袭和迁移能力。

Aim:To explore the effects and the molecular mechanism of lncRNA SNHG1 on the proliferation,invasion and migration of neuroblastoma SK-N-SH cells.Methods:SK-N-SH cells were transfected with SNHG1 shRNA(sh-SNHG1 group)or negative control shRNA(sh-NC group)by Lipofectamine2000,and the cells without transfection were the blank control.After 24 hours of culture,the expression of SNHG1 was detected by qRT-PCR,cell proliferation was detected by CCK-8 method,cell invasion and migration were detected by Transwell chamber test,and the expressions of MMP-9 and MMP-2 proteins were detected by Western blot.The binding sites of SNHG1 and miR-145 were predicted by online target gene prediction software Starbase,and the targeting relationship between them was identified by double luciferase report test.SNHG1 shRNA and miR-145 inhibitor were co-transfected into SK-N-SH cells(sh-SNHG1+anti-miR-145 group),and SNHG1 shRNA and inhibitor negative control were co-transfected into SK-N-SH cells(sh-SNHG1+anti-NC),then cell proliferation,invasion and migration were measured,as well as the expressions of miR-145,MMP-9 and MMP-2 proteins.Results:Compared with those of the blank control and sh-NC groups,the expressions of SNHG1,MMP-9 and MMP-2 proteins decreased in sh-SNHG1 group,and the proliferation,invasion and migration capabilities decreased(P<0.05).The results of double luciferase report showed that SNHG1 negatively regulated the expression of miR-145 in SK-N-SH cells.Compared with those of the sh-SNHG1+anti-NC group,the expression of miR-145 in sh-SNHG1+anti-miR-145 group was lower(P<0.05),the expressions of MMP-9 and MMP-2 proteins increased(P<0.05),and the proliferation,invasion and migration capabilities increased(P<0.05).Conclusion:SNHG1 could improve the proliferation,invasion and migration of neuroblastoma cells by downregulating the expression of miR-145.