南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (9): 1367-1373.doi: 10.12122/j.issn.1673-4254.2022.09.13

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RHPN2在骨肉瘤细胞中高表达并与较差的预后相关

刘智宇，方锋助，李 京，赵广跃，臧全金，张 峰，牒 军   

  1. 西安市人民医院（西安市第四医院）骨科病院，陕西 西安 710004；西安交通大学第二附属医院骨科，陕西 西安 710004
  • 发布日期:2022-09-28

RHPN2 is highly expressed in osteosarcoma cells to promote cell proliferation and migration and inhibit apoptosis

LIU Zhiyu, FANG Fengzhu, LI Jing, ZHAO Guangyue, ZANG Quanjin, ZHANG Feng, DIE Jun   

  1. Orthopedic Hospital of Xi'an People's Hospital (Xi'an Fourth Hospital), Xi'an 710004, China; Department of Orthopedics, Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, China
  • Published:2022-09-28

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摘要： 目的 筛选骨肉瘤细胞中异常表达的基因，探究RHPN2对骨肉瘤细胞体外增殖、凋亡和迁移能力以及对体内成瘤能力的影响。方法 采用GEO2R分析GSE70414数据集中骨肉瘤细胞与正常细胞的基因差异表达，RT-qPCR和Western blot实验检测人骨肉瘤细胞系MG-63、143B和SAOS2中RHPN2的表达情况。分别采用两种RHPN2-shRNA和对照组NC-shRNA转染143B细胞，CCK8实验、CFU实验、Annexin V-FITC/PI染色以及划痕试验分别检测RHPN2沉默对143B细胞增殖能力、凋亡和迁移能力的影响。采用143B细胞和RHPN2-shRNA转染的143B细胞原位注射构建裸鼠骨肉瘤原位瘤模型，并对肿瘤组织进行HE染色来验证RHPN2沉默对骨肉瘤细胞体内成瘤的影响。采用KM生存曲线分析RHPN2的表达与骨肉瘤患者生存预后的相关性。结果 RHPN2在骨肉瘤细胞系MG-63、143B和SAOS2中的表达明显上调（P<0.01）；RHPN2的沉默能够显著抑制骨肉瘤143B细胞的体外增殖和迁移，促进凋亡过程（P<0.01），并且抑制143B细胞的体内成瘤能力；RHPN2高表达的骨肉瘤患者的总体生存率明显低于低表达患者（P<0.05）。结论 RHPN2在骨肉瘤细胞中高表达，能够促进骨肉瘤细胞的增殖和迁移，抑制凋亡，并且与骨肉瘤患者较差的预后相关。

关键词: 骨肉瘤；生信分析；RHPN2；增殖；预后

Abstract: Objective To screen for aberrantly expressed genes in osteosarcoma cells and investigate the role of RHPN2 in regulating the proliferation, apoptosis, migration and tumorigenic abilities of osteosarcoma cells. Methods We used GEO2R to analyze the differential gene expression profile between osteosarcoma cells and normal cells in the GSE70414 dataset. RT-qPCR and Western blotting were performed to detect RHPN2 expression in osteosarcoma cell lines MG-63, 143B and SAOS2. Two RHPN2-shRNA and a control NC-shRNA were designed to silence the expression of RHPN2 in 143B cells, and CCK8 assay, colony-forming assay, annexin V-FITC/PI staining and scratch assays were carried out to examine the changes in proliferation, apoptosis and migration of the cells. We also established nude mouse models bearing osteosarcoma xenografts derived 143B cells and RHPN2-shRNA-transfected 143B cells, and assessed the effect of RHPN2 silencing on osteosarcoma cell tumorigenesis using HE staining. Kaplan-Meier survival curves were used to analyze the correlation between RHPN2 expression and survival outcomes of patients with osteosarcoma. Results RHPN2 expression was significantly upregulated in osteosarcoma cell lines MG-63, 143B and SAOS2 (P<0.01). Silencing of RHPN2 significantly inhibited the proliferation and migration of 143B cells in vitro, promoted cell apoptosis (P<0.01), and suppressed tumorigenic capacity of the cells in nude mice. A high expression of RHPN2 was significantly correlated with a poor prognosis of patients with osteosarcoma (P<0.05). Conclusion RHPN2 is highly expressed in osteosarcoma cells to promote cell proliferation and migration and inhibits cell apoptosis. A high expression of RHPN2 is associated with a poorer prognosis of the patients with osteosarcoma.

Key words: osteosarcoma; bioinformatics analysis; RHPN2; proliferation; prognosis