Effects of lentivirus vector-mediated Kruppel-like factor 5 transfection on proliferation and metastasis of RKO cells

Abstract

Objective To investigate the effects of Kruppel-like factor 5 ( KLF5 ) on biological behaviors of RKO cell line.Methods With total RNA extracted from human intestinal cells as the template,KLF5 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) with primers designed according to the sequence of GenBank,and the product was inserted into the pCDH-CMV-KLF5-EF1-copGFP to construct KLF5 lentivirus vector.RKO cells were divided into two groups:blank control group,KLF5 group. The synthesized KLF5 lentivirus vector was transfected into RKO cells. Real-time quantitative PCR and Western blotting were used to detect the KLF5 mRNA and protein levels.The proliferation of RKO cells was measured by cell counting Kit-8 (CCK-8) assay.The ability of motility and invasion of RKO cells was assessed by Transwell invasion assay.Results In the KLF5 group,the expression of KLF5 mRNA and protein was increased as compared with the blank control group ( P ＜ 0.05 ).The overexpression of KLF5 could significantly suppressed proliferation of RKO cells ( P ＜ 0.05 ).KLF5 could significantly inhibit the abilities of migration and invasion of RKO cells as compared with the blank control group ( 50.26 ± 2.17 vs 25.12 ± 2.27 ; 45.48 ± 1.53 vs 22.13 ± 2.25 ) (P＜0.05).Conclusion KLF5 could regulate the biological behaviors of RKO cell and be used as a target to prevent the colon cancer. Key words: Colon cancer; Lentivirus vector; Kruppel-like factor 5