Influence of the Twist gene on the invasion and metastasis of colon cancer

Abstract

The present study investigated the role of the Twist gene in epithelial-mesenchymal transition (EMT) and its effects on the invasion and metastasis of malignant tumors. In vitro, we transfected SW480, HCT116 and HT29 cells with recombinant plasmids, pTracer-CMV/BSD-Twist and pGenesil1.2-Twist-shRNA, to influence expression of Twist. The transfection efficacy of the plasmids in the cell lines was confirmed by flow cytometry. The relative mRNA and protein expression levels of Twist, E-cadherin and vimentin in the transfected cells were detected by RT-PCR and western blotting, respectively. In addition, migration and invasion were assessed by Transwell assays. In vivo, we established a xenogenic liver metastasis mouse model by intrasplenic injection with transfected SW480, HCT116 or HT29 human colon cancer cells and used hematoxylin and eosin (H&E) staining to demonstrate the effective establishment of the model. The relative mRNA levels of Twist and vimentin were detected by RT-PCR. In vitro, RT-PCR and western blotting showed higher relative mRNA and protein expression levels of Twist and vimentin in cell lines transfected with the recombinant, highly expressed Twist plasmid than in non-transfected cell lines (P<0.05), while E-cadherin was inhibited (P<0.05). After transfection with the plasmid pGenesil1.2-Twist-shRNA, the relative mRNA and protein levels of Twist and vimentin were markedly inhibited in the HCT116 cells (P<0.05), and the levels of E-cadherin were not changed (P>0.05), along with inhibition of the migration and invasion abilities of the cell line (P<0.01). In vivo, relative mRNA levels of Twist and vimentin in both the liver and spleen of the mouse model were higher in the groups that were injected with one of the three cell lines transfected with pTracer-CMV/BSD-Twist than in the groups injected with cells transfected with pGenesil1.2-Twist-shRNA (P<0.05). In conclusion, upregulation of Twist gene expression can promote EMT molecular events. Interfering with the Twist gene can effectively silence Twist gene expression in HCT116 cells and consequently inhibit colon cancer cell migration and invasion.

Knockdown of MMP‑1 inhibits the progression of colorectal cancer by suppressing the PI3K/Akt/c‑myc signaling pathway and EMT

“…A study by Satelli and Li provided evidence that vimentin promotes tumorigenesis via various signaling pathways (42). Twist is a typical gene in EMT, and its function of promoting the invasion and metastasis of colon cancer cells has been demonstrated by Wang et al (43). These results indicated that MMP-1 promoted invasion and metastasis not only by degrading the ECM but also by activating EMT.…”

Functions of chemokines in the perineural invasion of tumors (Review)

“…Upregulation of Twist was found in certain human tumors, including breast cancer (111), prostate cancer (112) and gastric cancer (113). Twist mainly functions by promoting EMT in malignant tumors (111,114).…”

Leptin Promotes Expression of EMT-Related Transcription Factors and Invasion in a Src and FAK-Dependent Pathway in MCF10A Mammary Epithelial Cells

“…In this study, we observed that β-catenin expression increased, and was located in the nucleus of leptin-stimulated cells for 3 h. These findings may correlate to the decrease in the expression or changes in the location of proteins from the adherent junctions, in particular, E-cadherin and β-catenin [90]. The leptin-induced overexpression of both TFs and their nuclear localization could be related to the expression of EMT marker genes, such as MMPs expression, which are essential for enhanced invasive capabilities of mammary epithelial cells MCF10A, as previously described [91,92,93].…”