Page 64 - 南京医科大学学报自然科学版

南京医科大学学报（自然科学版） 第43卷第4期
·502 · Journal of Nanjing Medical University（Natural Sciences） 2023年4月

·基础研究·

miR⁃27b⁃3p通过靶向SSRP1调控骨肉瘤细胞的生长

段伟豪，俞学成，吴璟斌，袁秀琛，范世杰，谭亚东，翁益平 *
南京医科大学附属常州市第二人民医院骨科，南京医科大学常州医学中心，江苏 常州 213003

［摘 要］ 目的：探究miR⁃27b⁃3p对骨肉瘤（osteosarcoma，OS）细胞生长的影响及其潜在的分子机制。方法：①采用实时荧光
定量PCR（RT⁃qPCR）分析miR⁃27b⁃3p在骨肉瘤细胞系中的表达；②CCK⁃8实验、集落形成实验及流式细胞术检测miR⁃27b⁃3p
以及SSRP1对骨肉瘤细胞生长的影响；③使用在线数据库预测miR⁃27b⁃3p的靶基因并进行筛选，使用双荧光素报告酶实验测
定miR⁃27b⁃3p与结构特异性识别蛋白1（structure⁃specific recognition protein⁃1，SSRP1）的关系；④干扰效率以及过表达效率使
用Western blot实验验证。结果：①miR⁃27b⁃3p在骨肉瘤细胞和临床样本中低表达，过表达miR⁃27b⁃3p会抑制骨肉瘤细胞的生
长；②生物信息学分析和双荧光素报告酶实验证实SSRP1为miR⁃27b⁃3p的靶基因，SSRP1表达受miR⁃27b⁃3p的直接调控；③干
扰 SSRP1 的表达会抑制骨肉瘤细胞的增殖，促进凋亡；④过表达 SSRP1 会部分逆转 miR⁃27b⁃3p 对骨肉瘤细胞生长的抑制效
应。结论：miR⁃27b⁃3p在骨肉瘤细胞中低表达，通过靶向SSRP1从而影响骨肉瘤细胞的生长。miR⁃27b⁃3p可能是骨肉瘤的一
个潜在的分子靶点，靶向miR⁃27b⁃3p/SSRP1轴可能成为骨肉瘤治疗的新策略。
［关键词］ 骨肉瘤；miR⁃27b⁃3p；SSRP1；细胞增殖；细胞凋亡
［中图分类号］ R730.231 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）04⁃502⁃08
doi：10.7655/NYDXBNS20230408

MiR⁃27b⁃3p inhibits osteosarcoma cells growth by targeting SSRP1

DUAN Weihao，YU Xuecheng，WU Jingbin，YUAN Xiuchen，FAN Shijie，TAN Yadong，WENG Yiping *
Department of Orthopedics，the Affiliated Changzhou Second People’s Hospital of Nanjing Medical University，
Changzhou Medical Center，Nanjing Medical University，Changzhou 213003，China

［Abstract］ Objective：To investigate the effect of miR⁃27b⁃3p on osteosarcoma cells growth and its potential molecular mechanism.
Methods：①The expression of miR⁃27b⁃3p in osteosarcoma cell lines was analyzed by RT⁃qPCR. ②CCK⁃8 assay，colony formation
assay and flow cytometry analysis were used to detect the effect of miR⁃27b⁃3p and structure⁃specific recognition protein⁃1（SSRP1）on
osteosarcoma cell growth. ③The miRNA target prediction database was used to predict potential target gene of miR⁃27b⁃3p，and dual
luciferase reporter assay was performed to demonstrate the relationship of miR⁃27b⁃3p and its target genes. ④Efficiency of knockdown
or overexpression was validated by Western blot analysis. Results：①The expression of miR⁃27b⁃3p in osteosarcoma cell lines and
clinical samples was low，and overexpression of miR⁃27b⁃3p inhibited the growth of osteosarcoma cells. ②Bioinformatics analysis and
dual⁃fluorescence reporter assay confirmed that SSRP1 was a target gene of miR⁃27b⁃3p，and the expression of SSRP1 was regulated by
miR⁃27b⁃3p. ③Silencing of SSRP1 significantly inhibited cell proliferation and increased cell apoptosis of osteosarcoma cells.
④Overexpression of SSRP1 partly reversed the inhibitory effect of miR⁃27b⁃3p on osteosarcoma cell growth. Conclusion：miR⁃27b⁃3p
is low expressed in osteosarcoma cells，and it regulates the growth of osteosarcoma cells by targeting SSRP1. Targeting the miR⁃27b⁃3p/
SSRP1 axis may become a new therapeutic strategy for the treatment of osteosarcoma.
［Key words］ osteosarcoma；miR⁃27b⁃3p；SSRP1；cell proliferation；cell apoptosis
［J Nanjing Med Univ，2023，43（04）：502⁃509］

［基金项目］ 常州市科技计划项目（CE20215020）；南京医科大学常州医学中心科研项目（CMCC202217）
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通信作者（Corresponding author），E⁃mail：czeywengyiping@163.com