Tang Xianglian,Liu Denghui,Li Ming,et al.Effect of mir-29b in the proliferation,invasion and migration of neuroblastoma cells[J].Journal of Clinical Pediatric Surgery,2020,19(08):711-716.[doi:10.3969/j.issn.1671-6353.2020.08.010]
miR-29b对神经母细胞瘤增殖、侵袭和转移的影响研究
- Title:
- Effect of mir-29b in the proliferation,invasion and migration of neuroblastoma cells
- Keywords:
- Neuroblastoma; Neoplasm Invasiveness; Neoplasm Metastasis
- 分类号:
- R739.4;R730.2
- 摘要:
- 目的 检测miR-29b在肾上腺和神经母细胞瘤组织中的表达情况，探讨miR-29b对神经母细胞瘤细胞增殖、侵袭和转移能力的影响。方法 收集12例神经母细胞瘤患儿的肿瘤组织和癌旁肾上腺组织标本，实时定量PCR检测miR-29b的表达水平；miR-29相关核苷酸（mimics、inhibitor、NC）瞬时转入SK-N-SH神经母细胞瘤细胞后，CCK-8实验检测miR-29b对该细胞系增殖的抑制作用，Transwell小室、划痕实验进行侵袭、迁移能力实验，Western blot方法检测上皮转化相关蛋白（E-cadherin、Vimentin）的表达水平。结果 与癌旁肾上腺组织比较，miR-29b在神经母细胞瘤组织的表达水平明显降低（P<0.05）；miR-29相关核苷酸（mimics、inhibitor、NC）转染细胞后，miR-29b mimics组出现抑制细胞增殖的效应（P<0.05）；Transwell小室实验显示miR-29b mimics组的穿膜细胞量（36.33±8.08）少于miR-29b inhibitor组（112.30±11.68）和NC组（112.30±11.68）；划痕实验显示转染miR-29b mimics后，其愈合率（51.67±4.50）%较miR-29b inhibitor组（87.67±3.79）%和NC组（85.00±3.00）%明显下降，细胞的侵袭、迁移能力明显下降；Western blot实验显示，与NC组相比，miR-29b mimics组E-cadherin表达增高，Vimentin表达降低，而miR-29b inhibitor组中E-cadherin表达降低，Vimentin表达增高（P<0.05）。结论 神经母细胞瘤组织中miR-29b表达水平下降，miR-29b过表达可以抑制细胞的增殖、侵袭和迁移能力，这种抑癌作用可能与抑制EMT有关。
- Abstract:
- Objective To detect the expression of miR-29b in adrenal gland and neuroblastoma and examine the effect of miR-29b on the proliferation, invasion and migration of neuroblastoma cells. Methods The expression of miR-29b was detected by real-time quantitative polymerase chain reaction (PCR) in neuroblastoma tissues and paracancerous adrenal tissues. And miR-29b related nucleotides (mimics/inhibitor/NC) were transiently transferred into neuroblastoma SK-N-SH cells. The growth inhibition of miR-29b on SK-N-SH was determined bv CCK-8 assay, cell migration and invasion ability was evaluated by Transwell invasion and Scratch test and the expressions of epithelial transformation related proteins (E-cadherin & vimentin) were detected by Western blot. Results miR-29b became down-regulated in neuroblastoma tissues compared with that in paracancerous adrenal tissues (P<0.05). After transfection with miR-29-related nucleotides (mimics/inhibitor/NC), miR-29b mimics exhibited the effect of inhibiting cell proliferation (P<0.05); Transwell tests showed that the migrated cells in miR-29b mimics group (36.33±8.08) were less than that in miR-29b inhibitor (112.30±11.68) and NC group (112.30±11.68); Scratch test showed that the healing rate (51.67±4.50) of mir-29b was significantly lower than that of mir-29b inhibitor group (87.67±3.79) and NC group (85.00±3.00). And migration and invasion ability of K-N-SH cells decreased significantly (P<0.05); Western blot showed that the expression of E-cadherin increased in miR-29b mimics group and the expression of Vimentin declined while the opposite result appeared in miRNA-29b inhibitor group (P<0.05). Conclusion miR-29b is down-regulated in neuroblastoma tissues. And an overexpression of miR-29b can inhibit cell proliferation, invasion and migration of neuroblastoma SK-N-SH cells. It may be related with an inhibition of epithelial mesenchymal transition (EMT).
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备注/Memo
收稿日期:2019-09-25。
基金项目:湖南省自然科学基金面上项目（编号：2018JJ2210）
通讯作者:李勇,Email:liyongpuwaike@163.com