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.
Objective: Transoral scarless thyroid surgery has proven to be a popular alternative to traditional approaches. Transoral robotic thyroidectomy (TORT) has been reported using ports on the lower lip and axilla. Avoiding axillary incision can further reduce scars on the armpit. Here, we present our preliminary data from the initial 20 consecutive patients to explore the feasibility of three-port TORT without axillary incision.Methods: From September 2017 to June 2019, we performed TORT at Beijing United Family Hospital using three intraoral ports without axillary incision via the da Vinci Si system with three robotic arms. The outcomes of the procedure were retrospectively reviewed.Results: Among 20 patients (mean age 30 ± 7 years; mean tumor size 1.64 ± 0.96 cm), 16 patients underwent unilateral thyroid lobectomy and four had total thyroidectomy with or without central neck dissection. Eighteen patients had papillary thyroid carcinomas (PTC), one had a follicular thyroid carcinoma, and one had a thyroid adenoma. The mean surgical time was 221 ± 68 min. The mean number of retrieved central lymph nodes in the PTC patients was 5.6 ± 5. There was no permanent vocal cord palsy or hypocalcemia postoperatively. One patient had transient vocal cord palsy, which resolved within 1 week. Paresthesia of the lower lip and the chin was observed in nine patients, and one patient had a first-degree burn of the skin flap due to the lens. Conclusion:Three-port TORT without axillary incision is feasible for selected patients and would be a potential alternative for remote-access thyroid surgery to avoid leaving scars on the neck or the armpit.
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