CircRNA circTNPO1 promotes the proliferation and metastasis of osteosarcoma by sponging miR-338-3p.
10.3760/cma.j.cn112152-20200529-00496
- Author:
Jian Hong LU
1
;
Xiao Wen HUANG
2
;
Guo Qiang ZHANG
3
;
Yan MA
4
;
Jun Xin CHEN
4
Author Information
1. Department of Clinical Laboratory and Pathology, China Coast Guard Hospital of the People's Armed Police Force, Jiaxing 314000, China.
2. Department of Clinical Laboratory, Jiaxing Hospital of Traditional Chinese Medicine, Jiaxing 314001, China.
3. Department of Internal Medicine, China Coast Guard Hospital of the People's Armed Police Force, Jiaxing 314000, China.
4. Department of Orthopedic Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 310020, China.
- Publication Type:Journal Article
- Keywords:
Cell metastasis;
Cell proliferation;
Osteosarcoma;
circTNPO1;
miR-338-3p
- MeSH:
Animals;
Bone Neoplasms/pathology*;
Cell Line, Tumor;
Cell Movement;
Cell Proliferation/genetics*;
Gene Expression Regulation, Neoplastic;
Humans;
Ki-67 Antigen/metabolism*;
Mice;
MicroRNAs/metabolism*;
Osteosarcoma/secondary*;
RNA, Circular/metabolism*;
Sincalide/metabolism*
- From:
Chinese Journal of Oncology
2022;44(9):968-974
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To explore the effects of circTNPO1 on the proliferation and metastasis of osteosarcoma (OS) by sponging miR-338-3p. Methods: The expression of circTNPO1 on osteoblasts and multiple OS cell lines were detected by qRT-PCR. CircTNPO1 stable knockdown 143B cell line was constructed by sh-circTNPO1. Cell count kit 8 (CCK-8) assay and wound healing assay were applied to evaluate the proliferation and metastasis of this cell. Luciferase reporter assay was used to explore the binding between circTNPO1 and miR-338-3p. In xenograft tumor model, miR-338-3p inhibitor or its control was injected into the circTNPO1 knockdown tumors. The weight and size of the tumors were evaluated and Ki-67 expression was detected by immunohistochemistry. Results: The RNA expression of circTNPO1 in OS cell lines U2OS, HOS, MG63, 143B, ZOS and ZOSM were 2.73±0.27, 3.18±0.54, 4.33±0.52, 5.75±0.65, 4.50±0.49 and 3.96±0.35, respectively, higher than 1.00±0.09 in hFOB1.19 (P<0.001). CCK-8 assay revealed that after 48 h and 72 h, the absorbance of sh-circTNPO1 #1 was 0.81±0.05 and 1.09±0.06, while sh-circTNPO1 #2 143B cells was 0.84±0.04 and 1.2±0.04, which were sharply reduced compared with the control (1.00±0.06 and 1.49±0.06, P<0.001); after 48 h and 72 h, the absorbance of 143B cells transfected with circTNPO1 #1 and miR-338-3p (0.92±0.06 and 1.32±0.07) were higher than those of cells transfected with sh-circTNPO1 cells and miR NC (0.92±0.06 and 1.32±0.07, P<0.050). Wound healing assay demonstrated that the 24 hour-migration rates of sh-circTNPO1 #1 and sh-circTNPO1 #2 cells were (24.43±2.15)% and (39.70±4.20)% respectively, which were significantly lower than that of the control [(56.51±3.27)%, P<0.010]; the migration rates of sh-circTNPO1 #1+ miR NC and sh-circTNPO1 #1+ miR-338-3p inhibitor were (26.70±2.21)% and (46.10±5.71)%, with a significant difference (P<0.005). In xenograft tumor model, the weight and size of tumors in control, sh-circTNPO1 #1+ miR NC and sh-circTNPO1 #1+ miR-338-3p inhibitor mice were (458.80±158.10) mg, (262.50±82.09) mg, (395.40±137.60) mg and (593.00±228.40) mm(2,) (203.30±144.20) mm(2,) (488.60±208.60) mm(2,) respectively. Compared with control, sh-circTNPO1 tumors were significantly smaller (P<0.01). Injection with miR-338-3p inhibitor significantly reversed both the weight and size of tumors (P<0.05). Conclusion: CircTNPO1 promotes the proliferation and metastasis of OS by sponging miR-338-3p, which could be a new target for OS treatments.
