Abstract
Study on Effect of Vitexicarpin on the Biological Behavior of Colorectal Cancer Cells by Circ_0000419/MicroRNA-224
Medical school of Yangzhou University, Yangzhou 225009, Jiangsu, 1Department of Radiotherapy oncology, Nanjing Benq medical center, Jiangsu, Nanjing 210019, 2Laboratory Department, Yangzhou Center for Disease Control and Prevention, Yangzhou 225009, Jiangsu, 3Medical school of Yangzhou University, Yangzhou Center for Disease Control and Prevention & Jiangsu Key Laboratory of Zoonosis, Jiangsu, Yangzhou 225009, China
Correspondence Address:
Guimei Kong, Medical school of Yangzhou University, Yangzhou Center for Disease Control and Prevention & Jiangsu Key Laboratory of Zoonosis, Jiangsu, Yangzhou 225009, China, E-mail: gmkong123456@163.com
To investigate the effect and molecular mechanism of vitexicarpin on the biological behavior of colorectal cancer cells is the objective of the study. The human epithelial cell line Caco-2 cells were divided into control group, low, middle and high dose groups of vitexicarpin, plasmid cloning DNA-circ_0000419 group, plasmid cloning DNA group, anti-microRNA-224 group, anti-microRNA-224-negative control group, vitexicarpin+small interfering-circ_0000419 group and vitexicarpin+microRNA-224 group. Cell activity was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. Colony formation number was detected by plate clone formation experiment. Apoptosis was detected by flow cytometry. Scratch test was used to detect the migration distance of cells. Transwell method was used to detect the number of invasive cells; protein expression was detected by western blot; the expression levels of circ_0000419 and microRNA-224 were detected by real-time fluorescence quantitative polymerase chain reaction; dual-luciferase reporter assay was used to detect the targeting relationship between circ_0000419 and microRNA-224. After treatment with low, middle and high doses of vitexicarpin, the viability of Caco- 2 cells decreased, the number of colony formation decreased, the apoptosis rate increased, the migration distance decreased, the number of invasive cells decreased, the expression level of E-cadherin increased, the expression level of N-cadherin decreased, the expression level of circ_0000419 increased and the expression level of microRNA-224 decreased (p<0.05). Overexpression of circ_0000419 or inhibition of miR-224 expression decreased Caco-2 cell viability, decreased colony formation, increased apoptosis rate, decreased cell migration distance, decreased invasive cell number, increased expression of E-cadherin and decreased expression of N-cadherin (p<0.05). Interference with circ_0000419 or overexpression of microRNA-224 reverses the effects of vitexicarpin on proliferation, apoptosis, migration and invasion of Caco-2 cells; circ_0000419 targets the regulation of microRNA-224 expression. Vitexicarpin can inhibit the malignant biological behavior of colorectal cancer cells by circ_0000419/microRNA-224.
