MiR-30c对骨肉瘤细胞生物学功能的影响研究#br#

doi:10.3969/j.issn.1007-3205.2020.06.009

河北医科大学学报

MiR-30c对骨肉瘤细胞生物学功能的影响研究#br#

新疆医科大学第六附属医院关节一科，新疆乌鲁木齐 830002

出版日期:2020-06-25 发布日期:2020-06-29
作者简介:舒莉（1982-），男，新疆乌鲁木齐人，新疆医科大学第六附属医院主治医师，医学硕士，从事骨关节疾病诊治研究。
基金资助:
新疆维吾尔自治区自然科学基金项目（2017D01C257）

The research on the biological functions of miR-30c in osteosarcoma cells#br#

First Department of Joint Surgery， Sixth Affiliated Hospital of Xinjiang Medical University， Urumqi 830002， China

Online:2020-06-25 Published:2020-06-29

摘要/Abstract

摘要： 目的探究微小RNA(microRNA，miR)-30c在骨肉瘤细胞中可能发挥的生物学功能。
方法荧光定量PCR检测6种骨肉瘤细胞系及人正常成骨细胞中miR-30c的表达水平，筛选目的细胞系；将细胞分为空白组、对照组及miR-30c过表达组，克隆形成和CCK-8实验检测miR-30c对骨肉瘤细胞体外增殖能力的影响；流式细胞术检测miR-30c对骨肉瘤细胞凋亡的影响；Transwell小室实验分析骨肉瘤细胞体外侵袭和迁移能力的改变；裸鼠皮下成瘤实验验证miR-30c在体内对骨肉瘤细胞增殖的影响。
结果 U2OS、Saos2、Mg63、HuO9、KIKU、143B 6种OS细胞系中miR-30c表达水平均明显低于hFOB1.19细胞，U2OS细胞中miR-30c表达水平最低（P＜0.05）。 miR-30c过表达组克隆数目明显少于对照组和空白组（P＜0.05）。随时间延长，各组细胞增殖能力呈升高趋势，但miR-30c过表达组细胞增殖能力明显低于空白组和对照组，组间、时点间、组间·时点间交互作用差异有统计学意义（P＜0.05）。过表达miR-30c组U2OS细胞凋亡率明显高于空白组和对照组（P＜0.05）。miR-30c过表达组细胞转染miR-30c mimic后侵袭细胞数和迁移细胞数明显低于空白组和对照组（P＜0.05）。裸鼠皮下接瘤4周后，miR-30c过表达组小鼠体内移植瘤体积和重量明显小于空白组和对照组(P＜0.05)。
结论 miR-30c可能作为抑癌基因参与骨肉瘤细胞U2OS的增殖及转移。

关键词: 骨肉瘤, 微RNAs, 生物学功能

Abstract: Objective To investigate the potential biological functions of miR-30c in osteosarcoma(OS) cells.
Methods RT-PCR was used to detect the expression of miR-30c in seven cell lines. The candidate cells were divided into 3 groups: the mock group, the negative group(NC) and miR-30c overexpression group(miR-30c mimic). Colony formation and CCK-8 assays were used to the detect the changes in cell proliferation after transfection. Flow cytometry was used to detect the effect of miR-30c on cell apoptosis in candidate OS cell. Transwell invasion and migration assays were conducted to assess the effects of miR-30c on cell metastasis capabilities in vitro after transfection. Subcutaneous tumor formation assay in nude mice was used to confirm the effect of miR-30c on OS cell proliferation in vivo.
Results The expression of miR-30c in U2OS, Saos2, Mg63, HuO9, KIKU, 143B cell lines were significantly lower than in hFOB1.19 while U2OS cells have the biggest fold changes with hFOB1.19(P＜0.05). The miR-30c mimic group had fewer cell clones than the mock group and NC group(P＜0.05). The cell proliferation ability of each group showed an increasing trend, the cell proliferation ability of miR-30c mimic group was significantly lower than the other groups, which had statistically significant differences in interactions between groups, time points and group·time points(P＜0.05). The apoptosis rate of U2OS cells in the miR-30c mimic was significantly higher than that in the mock group and NC group(P＜0.05). The capacity of cell invasion and migration were significantly suppressed in U2OS after transfection with miR-30c mimics when compared with NC group(P＜0.05). After 4 weeks of subcutaneous tumor implantation in nude mice, the volume and weight of transplanted tumors in miR-30c mimic group were significantly smaller than those in mock group and NC group(P＜0.05).
Conclusion miR-30c may be involved in the proliferation and metastasis of U2OS as a tumor suppressor gene.

Key words: osteosarcoma, microRNAs, biological functions

舒莉，巨啸晨，柴浩，杨德勇，孙荣鑫*. MiR-30c对骨肉瘤细胞生物学功能的影响研究#br#[J]. 河北医科大学学报, doi: 10.3969/j.issn.1007-3205.2020.06.009.

SHU Li, JU Xiao-chen, CHAI Hao, YANG De-yong, SUN Rong-xin*. The research on the biological functions of miR-30c in osteosarcoma cells#br#[J]. Journal of Hebei Medical University, doi: 10.3969/j.issn.1007-3205.2020.06.009.

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MiR-30c对骨肉瘤细胞生物学功能的影响研究#br#

The research on the biological functions of miR-30c in osteosarcoma cells#br#

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