Expression of miR-616 in osteosarcoma and its role in proliferation, apoptosis, migration and invasion of tumor cells

FU Wanlei; HAO Xianglin; CAO Ya; XIA Jiying; ZHOU Xiaogang; XU Jiayi; GUO Qiaonan

doi:10.16016/j.2097-0927.202508009

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Expression of miR-616 in osteosarcoma and its role in proliferation, apoptosis, migration and invasion of tumor cells

Basic Medicine | 更新时间：2025-12-25

- Expression of miR-616 in osteosarcoma and its role in proliferation, apoptosis, migration and invasion of tumor cells
- Journal of Army Medical University Vol. 47, Issue 20, Pages: 2461-2473(2025)
- 作者机构：
  
  陆军军医大学（第三军医大学）第二附属医院病理科，重庆
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(8197102971)
- DOI：10.16016/j.2097-0927.202508009
  CLC： R394.3;R730.23;R738.1
- Received：04 August 2025，
  
  Revised：2025-10-14，
  
  Accepted：16 October 2025，
  
  Published：30 October 2025
- 稿件说明：
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付万垒，郝翔麟，曹亚，等.MiR-616在骨肉瘤中的表达及其对细胞增殖、凋亡、迁移侵袭的作用［J］.陆军军医大学学报，2025，47（20）：2461-2473.DOI：10.16016/j.2097-0927.202508009.

FU W L，HAO X L，CAO Y，et al.Expression of miR-616 in osteosarcoma and its role in proliferation， apoptosis， migration and invasion of tumor cells［J］.J Army Med Univ，2025，47（20）：2461-2473.DOI：10.16016/j.2097-0927.202508009.
付万垒，郝翔麟，曹亚，等.MiR-616在骨肉瘤中的表达及其对细胞增殖、凋亡、迁移侵袭的作用［J］.陆军军医大学学报，2025，47（20）：2461-2473.DOI：10.16016/j.2097-0927.202508009. DOI：

FU W L，HAO X L，CAO Y，et al.Expression of miR-616 in osteosarcoma and its role in proliferation， apoptosis， migration and invasion of tumor cells［J］.J Army Med Univ，2025，47（20）：2461-2473.DOI：10.16016/j.2097-0927.202508009. DOI：

摘要

目的

阐释miR-616对骨肉瘤恶性生物学进程的影响，并初步探索其可能机制。

方法

采用原位杂交法（ISH）分析2018年1月至2019年12月陆军军医大学第二附属医院病理科11例骨肉瘤石蜡标本中miR-616表达；采用qRT-PCR比较成骨细胞hFOB1.19和骨肉瘤细胞系143B、HOS中miR-616的表达；通过慢病毒转染法构建miR-616敲低和过表达稳定细胞系，将细胞分为miR-616过表达组、过表达对照组、miR-616 敲低组和敲低对照组。采用流式细胞术检测各组细胞增殖和凋亡情况，采用Transwell和平板克隆形成实验检测各组细胞的侵袭和迁移情况。将10只4周龄雌性裸鼠（18~20 g）采用随机数字表法分为对照组和miR-616过表达组，采用皮下动物荷瘤实验比较两组移植瘤生长情况；采用二代测序和双萤光素酶报告系统确定miR-616的靶基因。

结果

ISH结果显示，与人骨肉瘤癌旁组织比较，miR-616在骨肉瘤组织中表达增高，主要存在于细胞质中，qRT-PCR结果表明miR-616在骨肉瘤细胞系143B、HOS中均高于成骨细胞hFOB1.19（

0.05）。体外实验表明，与过表达对照组比较，过表达miR-616组143B和HOS细胞的增殖能力增强（

0.01），凋亡抑制（

0.01），迁移能力增强（

0.01），侵袭能力增加（

0.01）；相反，敲低miR-616后，与敲低对照组比较，miR-616敲低组143B和HOS细胞增殖能力减弱（

0.05），迁移力减弱（

0.05），侵袭能力下降（

0.05），且miR-616-3p对骨肉瘤

的凋亡影响强于miR-616-5p。动物实验表明，过表达miR-616组的小鼠移植瘤质量（98.00±17.22）mg大于对照组（33.60±8.08）mg（

0.01）。二代测序和双荧光素酶报告系统结果显示，miR-616的靶蛋白为KLF2。

结论

miR-616可促进骨肉瘤恶性生物学行为发展，其表达水平可能成为骨肉瘤潜在治疗的靶点。

Abstract

Objective

To elucidate the effects of miR-616 on the malignant biological processes of osteosarcoma and to preliminarily explore its potential mechanisms.

Methods

In situ

hybridization (ISH) was employed to analyze miR-616 expression in 11 paraffin-embedded osteosarcoma specimens collected in our department during January 2018 to December 2019. Quantitative real-time PCR (qRT-PCR) was used to compare the mRNA expression level of miR-616 in the osteoblast cell line hFOB1.19 and osteosarcoma cell lines 143B and HOS. Stable cell lines with miR-616 knockdown or overexpression were established

via

lentiviral transfection in 143B and HOS cells. Cell proliferation and apoptosis were detected by flow cytometry

while cell invasion and migration were assessed using Transwell and colony formation assays

respectively. To evaluate the effect of miR-616 on tumor growth in

vivo

10 female nude mice (4 weeks old

weighing 18~20 g) were randomized into a control group and a miR-616 overexpression group. After the xenograft tumor model was constructed

the growth of subcutaneous tumors was monitored. Finally

next-generation sequencing and a dual-luciferase reporter assay were performed to identify the target genes of miR-616.

Results

ISH results showed that miR-616 expression was up-regulated in osteosarcoma tissues than adjacent tissues

and primarily localized in the cytoplasm. qRT-PCR confirmed that miR-616 level was significantly higher in 143B and HOS cells than hFOB1.19 cells (

0.05).

In vitro

experiments revealed that miR-616 overexpression enhanced the prol

iferation

migration and invasion

while suppressing apoptosis in 143B and HOS cells (

0.01). Conversely

miR-616 knockdown weakened these malignant phenotypes (

0.05)

with miR-616-3p showing a stronger effect on apoptosis than miR-616-5p. Animal experiments demonstrated that the tumor weight in the miR-616 overexpression group was significantly greater than that of the control group (98.00±17.22

33.60±8.08 mg

0.01). Furthermore

KLF2 was identified and confirmed as a direct target of miR-616.

Conclusion

MiR-616 promotes malignant biological behaviors in osteosarcoma

and its expression level indicates that it may serve as a potential therapeutic target.

关键词

Keywords

references

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