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

Lu,Sumei; Yu,Liang; Mu,Yakui; Ma,Juke; Tian,Jiajun; Xu,Wei; Wang,Haibo

doi:10.3892/mmr.2014.2212

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

Authors:
- Sumei Lu
- Liang Yu
- Yakui Mu
- Juke Ma
- Jiajun Tian
- Wei Xu
- Haibo Wang
View Affiliations

Affiliations: Department of Otorhinolaryngology Head and Neck Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong 250022, P.R. China
Published online on: May 6, 2014 https://doi.org/10.3892/mmr.2014.2212
Pages: 53-60
Copyright: © Lu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

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.

View Figures

View References

1	Mehta PS and Harrison LB: Function and organ preservation in adult cancers of the head and neck. Expert Rev Anticancer Ther. 7:361–371. 2007. View Article : Google Scholar : PubMed/NCBI
2	Vokes EE: Induction chemotherapy for head and neck cancer: recent data. Oncologist. 15(Suppl 3): 3–7. 2010. View Article : Google Scholar : PubMed/NCBI
3	Shukla S, Wu CP and Ambudkar SV: Development of inhibitors of ATP-binding cassette drug transporters: present status and challenges. Expert Opin Drug Metab Toxicol. 4:205–223. 2008. View Article : Google Scholar : PubMed/NCBI
4	Neyns B, Tosoni A, Hwu WJ and Reardon DA: Dose-dense temozolomide regimens: antitumor activity, toxicity, and immunomodulatory effects. Cancer. 116:2868–2877. 2010. View Article : Google Scholar : PubMed/NCBI
5	Baguley BC: Multiple drug resistance mechanisms in cancer. Mol Biotechnol. 46:308–316. 2010. View Article : Google Scholar : PubMed/NCBI
6	O’Connor R: The pharmacology of cancer resistance. Anticancer Res. 27:1267–1272. 2007.
7	Kruse AL and Grätz KW: Oral carcinoma after hematopoietic stem cell transplantation - a new classification based on a literature review over 30 years. Head Neck Oncol. 1:292009.PubMed/NCBI
8	Longley DB and Johnston PG: Molecular mechanisms of drug resistance. J Pathol. 205:275–292. 2005. View Article : Google Scholar : PubMed/NCBI
9	Dizdarevic S and Peters AM: Imaging of multidrug resistance in cancer. Cancer Imaging. 11:1–8. 2011. View Article : Google Scholar
10	Vasiliou V, Vasiliou K and Nebert DW: Human ATP-binding cassette (ABC) transporter family. Hum Genomics. 3:281–290. 2009. View Article : Google Scholar : PubMed/NCBI
11	Cianfriglia M, Cenciarelli C, Barca S, Tombesi M, Flego M and Dupuis ML: Monoclonal antibodies as a tool for structure-function studies of the MDR1-P-glycoprotein. Curr Protein Pept Sci. 3:513–530. 2002. View Article : Google Scholar : PubMed/NCBI
12	van den Heuvel-Eibrink MM, van der Holt B, Burnett AK, et al: CD34-related coexpression of MDR1 and BCRP indicates a clinically resistant phenotype in patients with acute myeloid leukemia (AML) of older age. Ann Hematol. 86:329–337. 2007.PubMed/NCBI
13	Kuo MT: Roles of multidrug resistance genes in breast cancer chemoresistance. Adv Exp Med Biol. 608:23–30. 2007. View Article : Google Scholar : PubMed/NCBI
14	Zhu K, Chen L, Han X and Wang J and Wang J: Short hairpin RNA targeting Twist1 suppresses cell proliferation and improves chemosensitivity to cisplatin in HeLa human cervical cancer cells. Oncol Rep. 27:1027–1034. 2012.PubMed/NCBI
15	Qin Q, Xu Y, He T, Qin C and Xu J: Normal and disease-related biological functions of Twist1 and underlying molecular mechanisms. Cell Res. 22:90–106. 2012. View Article : Google Scholar : PubMed/NCBI
16	Zhan X, Feng X, Kong Y, Chen Y and Tan W: JNK signaling maintains the mesenchymal properties of multi-drug resistant human epidermoid carcinoma KB cells through snail and twist1. BMC Cancer. 13:1802013. View Article : Google Scholar
17	Ma J, Lu S, Yu L, et al: FaDu cell characteristics induced by multidrug resistance. Oncol Rep. 26:1189–1195. 2011.PubMed/NCBI
18	Liang Y, Meleady P, Cleary I, McDonnell S, Connolly L and Clynes M: Selection with melphalan or paclitaxel (Taxol) yields variants with different patterns of multidrug resistance, integrin expression and in vitro invasiveness. Eur J Cancer. 37:1041–1052. 2001. View Article : Google Scholar
19	Laborde E: Glutathione transferases as mediators of signaling pathways involved in cell proliferation and cell death. Cell Death Differ. 17:1373–1380. 2010. View Article : Google Scholar : PubMed/NCBI
20	Yu L, Li HZ, Lu SM, et al: Alteration in TWIST expression: possible role in paclitaxel-induced apoptosis in human laryngeal carcinoma Hep-2 cell line. Croat Med J. 50:536–542. 2009. View Article : Google Scholar : PubMed/NCBI
21	Lemma S, Karihtala P, Haapasaari KM, et al: Biological roles and prognostic values of the epithelial-mesenchymal transition-mediating transcription factors Twist, ZEB1 and Slug in diffuse large B-cell lymphoma. Histopathology. 62:326–333. 2013. View Article : Google Scholar
22	Banerjee A, Qian P, Wu ZS, et al: Artemin stimulates radio- and chemo-resistance by promoting TWIST1-BCL-2-dependent cancer stem cell-like behavior in mammary carcinoma cells. J Biol Chem. 287:42502–42515. 2012. View Article : Google Scholar
23	Zhuo WL, Wang Y, Zhuo XL, Zhang YS and Chen ZT: Short interfering RNA directed against TWIST, a novel zinc finger transcription factor, increases A549 cell sensitivity to cisplatin via MAPK/mitochondrial pathway. Biochem Biophys Res Commun. 369:1098–1102. 2008. View Article : Google Scholar : PubMed/NCBI
24	Li L, Jiang AC, Dong P, Wang H, Xu W and Xu C: MDR1/P-gp and VEGF synergistically enhance the invasion of Hep-2 cells with multidrug resistance induced by taxol. Ann Surg Oncol. 16:1421–1428. 2009. View Article : Google Scholar : PubMed/NCBI
25	Saxena M, Stephens MA, Pathak H and Rangarajan A: Transcription factors that mediate epithelial-mesenchymal transition lead to multidrug resistance by upregulating ABC transporters. Cell Death Dis. 2:e1792011. View Article : Google Scholar : PubMed/NCBI
26	Wallerand H, Robert G, Pasticier G, et al: The epithelial-mesenchymal transition-inducing factor TWIST is an attractive target in advanced and/or metastatic bladder and prostate cancers. Urol Oncol. 28:473–479. 2010. View Article : Google Scholar : PubMed/NCBI
27	Piotrowska H, Kucinska M and Murias M: Biological activity of piceatannol: leaving the shadow of resveratrol. Mutat Res. 750:60–82. 2012. View Article : Google Scholar : PubMed/NCBI
28	Wyllie AH: “Where, O death, is thy sting?” A brief review of apoptosis biology. Mol Neurobiol. 42:4–9. 2010.
29	Tomiyama A, Tachibana K, Suzuki K, et al: MEK-ERK-dependent multiple caspase activation by mitochondrial proapoptotic Bcl-2 family proteins is essential for heavy ion irradiation-induced glioma cell death. Cell Death Dis. 1:e602010. View Article : Google Scholar
30	Qin H, Srinivasula SM, Wu G, Fernandes-Alnemri T, Alnemri ES and Shi Y: Structural basis of procaspase-9 recruitment by the apoptotic protease-activating factor 1. Nature. 399:549–557. 1999. View Article : Google Scholar : PubMed/NCBI
31	Shang L, Liu J, Zhu Q, et al: Gypenosides protect primary cultures of rat cortical cells against oxidative neurotoxicity. Brain Res. 1102:163–174. 2006. View Article : Google Scholar : PubMed/NCBI
32	Wang JQ, Chen Q, Wang X, et al: Dysregulation of mitochondrial calcium signaling and superoxide flashes cause mitochondrial genomic DNA damage in Huntington disease. J Biol Chem. 288:3070–3084. 2013. View Article : Google Scholar : PubMed/NCBI
33	Demirci S, Kutluhan S, Naziroğlu M, Uğuz AC, Yürekli VA and Demirci K: Effects of selenium and topiramate on cytosolic Ca(2+) influx and oxidative stress in neuronal PC12 cells. Neurochem Res. 38:90–97. 2013.PubMed/NCBI
34	Naziroglu M: Molecular role of catalase on oxidative stress-induced Ca(2+) signaling and TRP cation channel activation in nervous system. J Recept Signal Transduct Res. 32:134–141. 2012.
35	Zhang X, Wang Q, Ling MT, Wong YC, Leung SC and Wang X: Anti-apoptotic role of TWIST and its association with Akt pathway in mediating taxol resistance in nasopharyngeal carcinoma cells. Int J Cancer. 120:1891–1898. 2007. View Article : Google Scholar : PubMed/NCBI