Role and mechanism of Twist1 in modulating the chemosensitivity of FaDu cells

Abstract

Multidrug resistance (MDR) is one of the most important obstacles affecting the efficacy of chemotherapy treatments for numerous types of cancer. In the present study, we have demonstrated the possible function of Twist1 in the chemosensitivity of head and neck squamous cell carcinoma (HNSCC) and have identified that its mechanism maybe associated with MDR1/P-gp regulation. To investigate this, the hypopharyngeal cancer cell line, FaDu, and its MDR cell line induced by taxol, FaDu/T, were employed. Stable transfectants targeted to Twist1 overexpression and Twist1 silencing based on FaDu were also conducted. Morphological observation, flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), western blotting and laser scanning confocal microscope detection were utilized to detect the associations between Twist1 and the chemosensitivity of FaDu cells. Our results demonstrated that Twist1 and MDR1/P-gp were upregulated in FaDu/T cells in a MDR dose-dependent manner. The anti-apoptotic capabilities of FaDu/T cells were enhanced during MDR progression, with apoptosis-related proteins (Bcl-2, Bax, activated caspase-3 and caspase-9) changing to resist apoptosis. Twist1 overexpression decreased the sensitivity of cells to taxol as revealed by a significant increase in MDR1/P-gp and IC50 (P<0.05). This overexpression also enhanced the resistance to apoptosis, with apoptotic proteins changing to resist cell death, and inhibited Ca2+ release induced by taxol (P<0.05). Detections in Twist1 silencing cells also confirmed this result. This study provided evidence that alterations of Twist1 expression modulates the chemosensitivity of FaDu cells to taxol. Therefore, Twist1 knockdown may be a promising treatment regimen for advanced hypopharyngeal carcinoma patients with MDR.

The relationship between EMT, CD44high/EGFRlow phenotype, and treatment response in head and neck cancer cell lines

“…Interestingly, a high expression of the TWIST1 gene was found to correlate with a poor radioresponse, suggesting that there may be a link between a mesenchymal phenotype and radioresistance in HNSCC. This is in line with the finding that overexpression of Twist1 enhances the resistance to apoptosis in the human hypopharyngeal carcinoma cell line FaDu and with our previous study where cells of the mesenchymal‐like CD44 high /EGFR low subpopulation of HNSCC cell lines were shown to be less sensitive to radiotherapy than their epithelial counterparts . Moreover, in a meta‐analysis, Twist1 was shown to correlate with low differentiation, lymph node metastasis, and local recurrence, features that all may be associated with an increased EMT phenotype of the tumor .…”

Knockdown of TWIST enhances the cytotoxicity of chemotherapeutic drugs in doxorubicin-resistant HepG2 cells by suppressing MDR1 and EMT

“…TWIST expression is elevated in hepatocellular carcinoma, breast cancer, nasopharyngeal carcinoma and oral carcinoma (21,(23)(24)(25). Previous studies have revealed a novel function of TWIST, and it has been reported to be involved in the development of acquired chemoresistance in human cancer cells (15,16,26). In the present study, TWIST and MDR1 were expressed to a significantly higher level in liver tissues compared with in non-cancerous tissues.…”

“…Consistent with a previous report (14), it was revealed that TWIST is involved in the development of acquired drug resistance in human cancer cells. TWIST overexpression is also correlated with chemotherapy resistance in various types of cancer and leads to a poorer prognosis (15,16). Therefore, TWIST may be considered a novel therapeutic target in overcoming MDR in liver cancer.…”

Expression of α-fetoprotein in gastric cancer AGS cells contributes to invasion and metastasis by influencing anoikis sensitivity

“…Reverse transcription (RT)-PCR and sequencing. Total RNA was extracted and RT-PCR was conducted as previously described (10). The annealing temperature was 56-58˚C for AFP and 60˚C for β-actin amplification.…”