Mechanisms of miR-103a-3p/CHI3L1 in proliferation and vascular mimicry of ovarian cancer cells
10.3760/cma.j.cn371439-20191224-00031
- VernacularTitle:miR-103a-3p/CHI3L1在卵巢癌细胞增殖和血管拟生中的作用机制
- Author:
Lifen YANG
1
;
Wei SONG
;
Dawei XU
;
Jun WU
;
Ran GAO
Author Information
1. 唐山市妇幼保健院妇产科 063000
- From:
Journal of International Oncology
2020;47(6):333-339
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the mechanisms of microRNA (miR)-103a-3p/chitinase-3-like protein 1 (CHI3L1) in the proliferation and vascular mimicry of ovarian cancer cells and its effect on transforming growth factor-β (TGF-β) pathway.Methods:The relationship between the expression level of miR-103a-3p and the overall survival rate of ovarian cancer patients was analyzed by bioinformatics. The human ovarian adenocarcinoma SKOV3 cells were divided into 4 groups: control group, miR-103a-3p mimic group, miR-103a-3p mimic+ CHI3L1 group and CHI3L1 group. Quantitative polymerase chain reaction (qPCR) and Western blotting were used to detect the expression levels of miR-103a-3p, CHI3L1 mRNA and CHI3L1 protein respectively. The expression level of YKL-40 in cell culture fluid was detected by enzyme-linked immunosorbent assay. The cell viability, proliferation ability and angiogenesis ability of the 4 groups were detected by CCK-8 method, clone formation experiment and angiogenesis experiment. The dual luciferase report verified that miR-130a-3p targeted CHI3L1.Results:The overall survival rate of ovarian cancer patients with high expression of miR-103a-3p was higher than that of patients with low expression of miR-103a-3p ( χ2=6.187, P=0.048). The differences in miR-103a-3p and CHI3L1 mRNA levels among the control group, miR-103a-3p mimic group, miR-103a-3p mimic+ CHI3L1 group and CHI3L1 group were statistically significant ( F=198.254, P<0.001; F=60.214, P<0.001), miR-103a-3p mimic group and miR-103a-3p mimic+ CHI3L1 group had higher miR-103a-3p levels than the control group (all P<0.001), CHI3L1 group had higher CHI3L1 mRNA level than the control group ( P<0.001). The expression levels of CHI3L1 protein in the 4 groups were 2.25±0.23, 1.19±0.12, 2.29±0.28 and 4.31±0.37, and the difference was statistically significant ( F=18.675, P<0.001). The expression levels of YKL-40 in the cell culture fluids of the 4 groups were (1.84±0.20) ng/ml, (0.95±0.08) ng/ml, (2.64±0.25) ng/ml, (6.27±0.79) ng/ml, and the difference was statistically significant ( F=35.297, P<0.001). The YKL-40 level of the CHI3L1 group was significantly higher than that of the control group ( P<0.001), the miR-103a-3p mimic group was lower than the control group ( P<0.001), and the miR-103a-3p mimic+ CHI3L1 group was higher than the miR-103a-3p mimic group ( P<0.001). The cell viabilities of the 4 groups were 100%±2.54%, 76.23%±2.13%, 104.89%±3.56% and 137.42%±2.80%, and the difference was statistically significant ( F=23.584, P<0.001). The cell viability of the miR-103a-3p mimic group was significantly lower than that of the control group ( P<0.001), the CHI3L1 group was higher than the control group ( P<0.001), and the miR-103a-3p mimic+ CHI3L1 group was higher than the miR-103a-3p mimic group ( P<0.001). The number of clones formed in the 4 groups were 76.85±4.67, 21.56±2.85, 72.06±5.07 and 169.63±9.21, and the difference was statistically significant ( F=31.541, P<0.001). The proliferation capacity of the miR-103a-3p mimic group was significantly lower than that of the control group ( P<0.001), the CHI3L1 group was higher than the control group ( P<0.001), and the miR-103a-3p mimic+ CHI3L1 group was significantly higher than the miR-103a-3p mimic group ( P<0.001). The differences in the relative tube lengths and the tube bramches of the 4 groups were both statistically significant ( F=24.254, P<0.001; F=27.564, P<0.001). The differences in TGF-β and Smad levels of the 4 groups were both statistically significant ( F=30.254, P<0.001; F=34.187, P<0.001). The results of dual luciferase experiments showed that compared with the NC group, the luciferase activity in cells co-transfected of miR-103a-3p and CHI3L1-wt was significantly reduced. The difference of luciferase activity between the cells transfected with NC and co-transfected with miR-103a-3p and CHI3L1-mut was not significant. Conclusion:MiR-103a-3p can directly inhibit the expression of CHI3L1 and inhibit the proliferation and angiogenesis of ovarian cancer SKOV3 cells to inhibit ovarian lymphatic metastasis and distant metastasis, which may be related to the TGF-β pathway.
