Mechanism of lncRNA MIR4435-2HG targeting miR-376a-3p to regulate the biological behavior of cholangiocarcinoma cells

LIU Wen-dong; ZHANG Jia-lin; YU Zi-dan

doi:10.11659/jjssx.12E022138

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Mechanism of lncRNA MIR4435-2HG targeting miR-376a-3p to regulate the biological behavior of cholangiocarcinoma cells

Basic Research | 更新时间：2024-01-22

- Mechanism of lncRNA MIR4435-2HG targeting miR-376a-3p to regulate the biological behavior of cholangiocarcinoma cells
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 33, Issue 1, Pages: 30-35(2024)
- 作者机构：
  
  1.惠州市第三人民医院肿瘤科，广东惠州 516001
  2.惠州市第三人民医院康复中心，广东惠州 516001
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.12E022138
  CLC： R735.7
- Published：25 January 2024，
  
  Received：27 December 2022，
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刘文东,张嘉麟,余紫丹.lncRNA MIR4435-2HG靶向miR-376a-3p调控胆管癌细胞生物学行为的机制研究[J].局解手术学杂志,2024,33(01):30-35.

LIU Wen-dong,ZHANG Jia-lin,YU Zi-dan.Mechanism of lncRNA MIR4435-2HG targeting miR-376a-3p to regulate the biological behavior of cholangiocarcinoma cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(01):30-35.
刘文东,张嘉麟,余紫丹.lncRNA MIR4435-2HG靶向miR-376a-3p调控胆管癌细胞生物学行为的机制研究[J].局解手术学杂志,2024,33(01):30-35. DOI： 10.11659/jjssx.12E022138.

LIU Wen-dong,ZHANG Jia-lin,YU Zi-dan.Mechanism of lncRNA MIR4435-2HG targeting miR-376a-3p to regulate the biological behavior of cholangiocarcinoma cells[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2024,33(01):30-35. DOI： 10.11659/jjssx.12E022138.

摘要

目的

探讨长链非编码RNA（lncRNA）MIR4435-2HG（MIR4435-2HG）对胆管癌细胞增殖、迁移、侵袭、凋亡的影响及其对微小RNA-376a-3p（miR-376a-3p）的调控作用。

方法

qRT-PCR法检测人肝内胆管上皮细胞HIBEpic与人胆管癌细胞RBE中MIR4435-2HG、miR-376a-3p的表达。将si-NC、si-MIR4435-2HG、miR-NC、miR-376a-3p mimics、si-MIR4435-2HG联合anti-miR-NC、si-MIR4435-2HG联合anti-miR-376a-3p分别转染至RBE细胞，作为si-NC组、si-MIR4435-2HG组、miR-NC组、miR-376a-3p组、si-MIR4435-2HG+anti-miR-NC组、si-MIR4435-2HG+anti-miR-376a-3p组；采用MTT法、Transwell小室法及流式细胞仪分别检测细胞增殖、迁移、侵袭及凋亡情况；双荧光素酶报告基因实验验证MIR4435-2HG与miR-376a-3p的靶向关系。Western blot检测相关蛋白表达。

结果

RBE细胞中MIR4435-2HG表达量升高（

0.05），miR-376a-3p表达量降低（

0.05）。与si-NC组比较，si-MIR4435-2HG组MIR4435-2HG表达、细胞活力及CyclinD1、MMP-2、MMP-9蛋白水平降低（

0.05），迁移及侵袭细胞数减少（

0.05），细胞凋亡率升高（

0.05）；与miR-NC组比较，miR-376a-3p组细胞活力及CyclinD1、MMP-2、MMP-9蛋白水平降低（

0.05），迁移及侵袭细胞数减少（

0.05），miR-376a-3p表达、细胞凋亡率升高（

0.05）。MIR4435-2HG可靶向调控miR-376a-3p；与si-MIR4435-2HG+anti-miR-NC组比较，si-MIR4435-2HG+anti-miR-376a-3p组细胞活力及CyclinD1、MMP-2、MMP-9蛋白水平升高（

0.05），迁移及侵袭细胞数增多（

0.05），miR-376a-3p表达、细胞凋亡率降低（

0.05）。

结论

敲低MIR4435-2HG可通过靶向调控miR-376a-3p进而抑制RBE细胞增殖、迁移、侵袭，并诱导其凋亡。

Abstract

Objective

To explore the effects of long non-coding RNA （lncRNA） MIR4435-2HG （MIR4435-2HG） on the proliferation， migration， invasion and apoptosis of cholangiocarcinoma cells and its regulatory effect on microRNA-376a-3p （miR-376a-3p）.

Methods

qRT-PCR method was used to detect the expression of MIR4435-2HG and miR-376a-3p in human intrahepatic bile duct epithelial cells HIBEpic and human cholangiocarcinoma cells RBE. si-NC， si-MIR4435-2HG， miR-NC， miR-376a-3p mimics， si-MIR4435-2HG and anti-miR-NC，and si-MIR4435-2HG and anti-miR-376a-3p were transfected into RBE cells， respectively，as the si-NC group， the si-MIR4435-2HG group， the miR-NC group， the miR-376a-3p group， the si-MIR4435-2HG+anti-miR-NC group， the si-MIR4435-2HG+anti-miR-376a-3p group. MTT method， Transwell chamber method and flow cytometry were used to detect cell proliferation， migration， invasion and apoptosis； dual luciferase reporter gene assay was used to verify the targeting relationship between MIR4435-2HG and miR-376a-3p.Western blot was used to detect the expression of related proteins.

Results

The expression of MIR4435-2HG was increased in RBE cells， while the expression of miR-376a-3p was decreased （

0.05）. Compared with the si-NC group， the MIR4435-2HG expression， cell viability， and protein levels of CyclinD1

MMP-2

MMP-9 in the si-MIR4435-2HG group were reduced （

0.05）， the numbers of migrating and invading cells were reduced （

0.05）， while the MIR4435-2HG expression and apoptosis rate were increased （

0.05）. Compared with the miR-NC group， the cell viability and protein levels of CyclinD1

MMP-2

MMP-9 in the miR-376a-3p group were decreased （

0.05）， the numbers of migrating and invading cells were decreased （

0.05）， while the MIR4435-2HG expression and apoptosis rate were increased （

0.05）. MIR4435-2HG was of targeted regulation on miR-376a-3p. Compared with the si-MIR4435-2HG+anti-miR-NC group， the cell viability and protein levels of CyclinD1

MMP-2

MMP-9 in the si-MIR4435-2HG+anti-miR-376a-3p group were increased （

0.05）， the numbers of migrating and invading cells were increased （

0.05）， while the MIR4435-2HG expression and apoptosis rate were decreased （

0.05）.

Conclusion

Knockdown of MIR4435-2HG can inhibit the proliferation， migration， invasion and induce apoptosis of RBE cells by targeting miR-376a-3p.

关键词

lncRNA MIR4435-2HGmiR-376a-3p胆管癌细胞增殖迁移侵袭

Keywords

lncRNA MIR4435-2HGmiR-376a-3pcholangiocarcinomacell proliferationmigrationinvasion

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