Effect of CXCL1 gene expression on the biological function of epithelial ovarian carcinoma cells in vitro.
- Author:
Wei ZHANG
1
;
Ning HUANG
;
Qi WANG
;
Zhi-jun YANG
;
Li LI
Author Information
- Publication Type:Journal Article
- MeSH: Cell Cycle; Cell Line, Tumor; Cell Movement; Cell Proliferation; Chemokine CXCL1; genetics; metabolism; Female; Gene Expression Regulation, Neoplastic; Humans; Lentivirus; genetics; Neoplasm Invasiveness; Neoplasms, Glandular and Epithelial; genetics; metabolism; pathology; Ovarian Neoplasms; genetics; metabolism; pathology; Plasmids; RNA, Messenger; metabolism; Recombinant Proteins; genetics; metabolism; Transfection
- From: Chinese Journal of Oncology 2013;35(2):109-113
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of CXCL1 gene expression on biological function of ovarian cancer cells and its mechanism of action.
METHODSRT-PCR was used to amplify the full-length sequence of CXCL1, which was used to construct the recombinant lentiviral plasmids, and then the plasmids were packaged with human renal epithelial cell line 293T cells, and the ovarian cancer 2780 cells were transfected with CXCL1 gene by virus supernatant particles. The expression of CXCL1 mRNA and protein in 2780 cells with lentivirus-mediated CXCL1expression were determined by RT-PCR and ELISA, respectively. The growth curve, cell cycle and colony formation in vitro were measured by MTT assay, flow cytometry and colony formation counting, respectively. The cell invasion and migration in vitro was detected by Transwell chambers and migration attack methods, respectively.
RESULTSThe sequencing results showed that the recombinant lentiviral plasmid of CXCL1-pWPI had 100% homology with the CXCL1 gene, identified by BLAST analysis. RT-PCR and ELISA results showed positive expression of CXCL1 mRNA and protein in the CXCL1-PWPI group. The doubling time of the CXCL1-PWPI group was significantly shorter than that in the A2780 and PWPI groups (P < 0.05). The rate of colony-formation in the CXCL1-PWPI group was also significantly higher than that of the A2780 and PWPI groups (P < 0.05). The proportion of cells in G1 phase (44.0%) in the CXCL1-PWPI group was also significantly lower compared with that in the PWPI and A2780 groups (61.4% and 62.7%), with a significant difference (P < 0.001). There was no significant difference between the cell migration capacity (P > 0.05) of the CXCL1-PWPI, PWPI and A2780 groups, but the invasion capacity of the CXCL1-PWPI group was significantly higher than that of the PWPI and A2780 groups (P < 0.05).
CONCLUSIONThe over-expression of CXCL1 gene may promote the proliferation and invasion ability of A2780 cells in vitro.
