miR-449b-5p inhibits ovarian cancer cell growth and cell cycle progression by targeting Cyclin E2
DOI: 10.3872/j.issn.1007-385x.2021.02.007
- VernacularTitle:miR-449b-5p 通过靶向Cyclin E2 抑制卵巢癌细胞生长和细胞周期进展
- Author:
LI Lingling
1
,
2
,
3
;
WANG Yujue
1
,
2
,
3
;
YUE Jun
1
,
2
,
3
;
MEI Jie
1
,
2
,
3
;
ZHAO Xiaolan
1
,
2
,
3
Author Information
1. Department of Obstetrics and Gynecology, Sichuan Academy of Medical Sciences &
2. Sichuan Provincial People'
3. s Hospital, Chengdu 610027, Sichuan, China
- Publication Type:Clinical Trial
- Keywords:
miR-449b-5p;
Cyclin E2;
ovarian cancer;
growth;
cell cycle
- From:
Chinese Journal of Cancer Biotherapy
2021;28(2):143-150
- CountryChina
- Language:Chinese
-
Abstract:
[Abstract] Objective: To investigate the effect and mechanism of miR-449b-5p on the proliferation of ovarian cancer cells. Methods:
Cancer tissue and corresponding para-cancerous tissue specimens from 20 patients who underwent surgery in the Department of
Obstetrics and Gynecology of Sichuan Provincial People's Hospital from June 2018 to June 2020 were collected for this study; in
addition, normal ovarian epithelial cell line (HOSEpiC) and six human cervical cancer cell lines (SKOV3, ES-2, OVCAR-3, HO8910,
CaOV-3 and A2780) were also selected. mRNA expressions of miR-449b-5p and CCNE2 in ovarian cancer tissues and cells were
detected by qPCR. The plasmids miR-NC, miR-499b-5p mimic, miR-499b-5p inhibitor and pc-CCNE2 were transfected into SKOV3
cells separately or in combination. Cell growth and cell cycle were measured by the CCK-8 method and Flow cytometry, the expression
of CCNE2 protein was detected by WB assay, respectively. The targeting relationship between miR-449b-5p and CCNE2 was verified
by Dual luciferase reporter assay. miR-499b-5p transfected SKOV3 cells were injected subcutaneously in nude mice to construct
xenograft model, and the tumor volume was measured weekly. Nude mice were sacrificed at day 42. The weight of the subcutaneous
tumors was weighed by an electronic balance, and the expressions of CCNE2 and Ki67 were detected by immunohistochemistry.
Results: Compared with normal ovarian tissues and epithelial cell line HOSEpiC, miR-499b expression was significantly downregulated
in human cervical cancer tissues and cell lines SKOV3, ES-2, OVCAR-3, HO8910, CaOV-3 and A2780 (P<0.01). Compared
with the Control group, the proliferation of SKOV3 cells in the miR-499b mimic group was significantly reduced (P<0.01) and the cell
proportion in G0/G1 phase was significantly increased ( P<0.01); while the proliferation of SKOV3 cells in the miR-499b inhibitor
group was significantly increased (P<0.01) and the cell proportion in G0/G1 phase was significantly reduced (P<0.01). Over-expression
of miR-499b-5p significantly inhibited the luciferase activity of wild type CCNE2 plasmid (P<0.01) but had no effect on the luciferase activity of the mutant CCNE2 plasmid. Compared with the miR-499b mimic group, the growth of SKOV3 cells in the miR-499b
mimic+pc-CCNE2 group was significantly increased (P<0.01) and the cell proportion in G0/G1 phase was significantly reduced
(P<0.01). Compared with the miR-NC group, the tumor volume and weight of nude mice in the miR-499b mimic group were
significantly reduced (all P<0.01), and the proportion of CCNE2 and Ki67 positive cells was significantly decreased (P<0.01).
Conclusion: miR-449b-5p inhibits the growth and cell cycle progression of ovarian cancer cells by targeting Cyclin E2.
- Full text:zgzlswzlzz-2021-2-143.pdf
