Effect of RBP2 gene silencing on proliferation, migration and invasion of ovarian epithelial cancer SKOV3/DDP cells and its mechanism.
10.3760/cma.j.cn112152-20200612-00555
- Author:
Tong Fu FENG
1
;
Dong Mei YAO
1
;
Rong ZHENG
1
;
Duo Sheng JIANG
2
;
Ling Li ZHANG
1
;
Qi XING
1
;
Li LI
3
Author Information
1. Department of Gynecology, Maternal and Child Health Hospital of Hubei Province, Wuhan 430070, China.
2. Department of Traditional Chinese Medicine, Maternal and Child Health Hospital of Hubei Province, Wuhan 430070, China.
3. Key Laboratory of High-incidence Tumor Prevention and Treatment, Ministry of Education, Guangxi Medical University, Nanning 530021, China.
- Publication Type:Journal Article
- Keywords:
Epithelial-mesenchymal transformation;
Invasion;
Migration;
Ovarian neoplasms;
Retinol-binding protein 2
- MeSH:
Animals;
Apoptosis;
Carcinoma, Ovarian Epithelial/genetics*;
Cell Line, Tumor;
Cell Proliferation;
Cisplatin/pharmacology*;
Drug Resistance, Neoplasm/genetics*;
Female;
Gene Silencing;
Humans;
Mice;
Mice, Nude;
Ovarian Neoplasms/pathology*;
Retinol-Binding Proteins, Cellular/metabolism*
- From:
Chinese Journal of Oncology
2022;44(2):139-146
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To explore the effect of down-regulation of retinol binding protein 2 (RBP2) expression on the biological characteristics of ovarian cancer cells and its mechanism. Methods: Knockdown of RBP2 and cisplatin (DDP)-resistant ovarian cancer cell line SKOV3/DDP-RBP2i was established, the negative control group and blank control group were also set. Cell counting kit 8 (CCK-8) was used to detect the cell proliferation ability, flow cytometry was used to detect cell apoptosis, scratch test and Transwell invasion test were used to detect cell migration and invasion ability, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) and western blot were used to detect the expressions of molecular markers related to epithelial-mesenchymal transition (EMT). The effect of RBP2 on the growth of ovarian cancer was verified through experiment of transplanted tumors in nude mice, and the relationships between RBP2 expression and tumor metastasis and patient prognosis were analyzed using the clinical data of ovarian cancer in TCGA database. Results: After down-regulating the expression of RBP2, the proliferation ability of SKOV3/DDP cell was significantly reduced. On the fifth day, the proliferation activities of SKOV3/DDP-RBP2i group, negative control group and blank control group were (56.67±4.16)%, (84.67±3.51) and (87.00±4.00)% respectively, with statistically significant difference (P<0.001). The apoptosis rate of SKOV3/DDP-RBP2i group was (14.19±1.50)%, higher than (8.77±0.75)% of the negative control group and (7.48±0.52)% of the blank control group (P<0.001). The number of invasive cells of SKOV3/DDP-RBP2i group was (55.20±2.39), lower than (82.60±5.18) and (80.80±7.26) of the negative control group and the blank control group, respectively (P<0.001). The scratch healing rate of SKOV3/DDP-RBP2i group was (28.47±2.72)%, lower than (50.58±4.06)% and (48.92±4.63)% of the negative control group and the blank control group, respectively (P<0.001). The mRNA and protein expressions of E-cadherin in the SKOV3/DDP-RBP2i group were higher than those in the negative control group (P=0.015, P<0.001) and the blank control group (P=0.006, P<0.001). The mRNA and protein expression of N-cadherin in SKOV3/DDP-RBP2i group were lower than those in the negative control group (P=0.012, P<0.001) and the blank control group (P=0.005, P<0.001). The mRNA and protein expressions of vimentin in SKOV3/DDP-RBP2i group were also lower than those in the negative control group (P=0.016, P=0.001) and the blank control group (P=0.011, P=0.001). Five weeks after the cells inoculated into the nude mice, the tumor volume of SKOV3/DDP-RBP2i group, negative control group and blank control group were statistically significant different. The tumor volume of SKOV3/DDP-RBP2i group was smaller than those of negative control group and blank control group (P=0.001). Bioinformatics analysis showed that the expression of RBP2 in patients with metastatic ovarian cancer was higher than that without metastasis (P=0.043), and the median overall survival of ovarian cancer patients with high RBP2 expression was 41 months, shorter than 69 months of low RBP2 expression patients (P<0.001). Conclusion: Downregulation of the expression of RBP2 in SKOV3/DDP cells can inhibit cell migration and invasion, and the mechanism may be related to the inhibition of EMT.
