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长链非编码 RNA-H19 基因在调控神经胶质瘤细胞功能方面发挥着重要作用。

LncRNA-H19 gene plays a significant role in regulating glioma cell function.

机构信息

Department of Oncology, Heping Hospital Affiliated to Changzhi Medical College, Changzhi, China.

出版信息

Mol Genet Genomic Med. 2021 Oct;9(10):e1480. doi: 10.1002/mgg3.1480. Epub 2021 Sep 3.

Abstract

BACKGROUND

Glioma is an aggressive adult primary cancer, and is characterized by low cure rate, poor prognosis, and high recurrence. The present study aimed to investigate the effect of lncRNA-H19 gene silencing on glioma cell function.

METHODS

lncRNA-H19 interference vector (LV3-si-H19) and negative control vector (LV3-NC) were stably transfected into U251 and U87-MG cells, respectively. Quantitative real-time PCR (qRT-PCR) was performed to investigate the expression of lncRNA-H19. Cell proliferation capacity was tested by adopting cell counting kit (CCK8), and propidium iodide (PI) was used for cell cycle analysis. Meanwhile, flow cytometry (FCM) method was used to investigate cell apoptosis, cell migration capacity was detected via wound healing and transwell experiments, and sphere-forming ability was examined in serum-free suspension culture. Additionally, glioma animal models were conducted through injecting U251 cells to estimate the effects of lncRNA-H19 on glioma growth in vivo.

RESULTS

Knocking down lncRNA-H19 gene could effectively suppress the proliferation of U251 and U87-MG cells. The knockdown of lncRNA-H19 remarkably inhibited the migration and blocked cycle progressions of U251 and U87-MG cells, yet, no obvious changes were observed in cell apoptosis. Besides, inhibiting lncRNA-H19 expression could attenuate sphere-forming function of U251 and U87-MG cells. Additionally, tumor volume and weight were significantly reduced in rats injected with U251 LV-si-H19 cell line compared to untransfected and negative controls, when survival time was obviously prolonged in U251 LV-si-H19 injection groups.

CONCLUSION

LncRNA-H19 gene plays a carcinogenic role in glioma progression via enhancing aggressive behavior of glioma cells.

摘要

背景

脑胶质瘤是一种侵袭性的成人原发性癌症,其特点是治愈率低、预后差、复发率高。本研究旨在探讨长链非编码 RNA-H19 基因沉默对脑胶质瘤细胞功能的影响。

方法

将 lncRNA-H19 干扰载体(LV3-si-H19)和阴性对照载体(LV3-NC)分别稳定转染至 U251 和 U87-MG 细胞中,采用实时定量 PCR(qRT-PCR)检测 lncRNA-H19 的表达。采用细胞计数试剂盒(CCK8)检测细胞增殖能力,碘化丙啶(PI)进行细胞周期分析。同时,采用流式细胞术(FCM)检测细胞凋亡,划痕愈合和 Transwell 实验检测细胞迁移能力,无血清悬浮培养检测球体形成能力。另外,通过注射 U251 细胞构建脑胶质瘤动物模型,评估 lncRNA-H19 对体内脑胶质瘤生长的影响。

结果

敲低 lncRNA-H19 基因可有效抑制 U251 和 U87-MG 细胞的增殖。敲低 lncRNA-H19 显著抑制 U251 和 U87-MG 细胞的迁移并阻断细胞周期进程,但对细胞凋亡无明显影响。此外,抑制 lncRNA-H19 表达可减弱 U251 和 U87-MG 细胞的球体形成功能。另外,与未转染组和阴性对照组相比,注射 U251 LV-si-H19 细胞系的大鼠肿瘤体积和重量明显减小,而 U251 LV-si-H19 注射组的生存时间明显延长。

结论

lncRNA-H19 基因通过增强脑胶质瘤细胞的侵袭行为在脑胶质瘤进展中发挥致癌作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e083/8580082/f787b293852c/MGG3-9-e1480-g002.jpg

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