Study on epithelial-mesenchymal transition induced by TGF-β1 enhances cisplatin-resistance capacity of human tongue squamous cell carci

doi:10.3877/cma.j.issn.1674-1366.2012.03.002

Abstract

Abstract:

Objective

The aim of this study was to establish an Epithelial-mesenchymal Transition model of oral squamous cell carcinoma cell line CAL27 and cisplatin-resistant cell line CAL27-res with TGF-β1 and investigate the effect of Epithelial-mesenchymal Transition on cisplatinresistance capacity， cell growth， and apoptosis.

Methods

Cultured CAL27 and CAL27-res cells were treated with 10 ng/ml TGF-β1 for 8 days. The morphological changes were observed by microscope； the changes of EMT relative marker proteins were assessed by Western blot and immunoflurescence staining.In addition， Pirquet test was used to investigate the change of motility in CAL27 and CAL27-res. A MTT-based method was used to analyze the half maximal inhibitory concentration （IC50） values.Immunofluorescence staining was used to investigate the expression of MDR， MRP and TopoⅡβ. Flow cytometry was used to analyze the growth and apoptosis.

Results

The results showed that TGF-β1 could induce CAL27 and CAL27-res cells morphological alteration from epithelial to mesenchymal and downregulate the expression of epithelial maker protein E-cadherin， upregulate the expression of mesenchymal maker protein Vimentin. Furthermore the motility was significantly enhanced. The IC50 of CAL27 and CAL27-res increased by 2.31 and 1.72 fold accompanied by MDR， MRP up-regulation and Topo Ⅱβ down-regulation. The Proliferation index （PI） of CAL27 and CAL27-res were significantly decreased （CAL27： χ²=16.799， P＜0.001； CAL27-res： χ²=25.375， P＜0.001）. The Apoptosis Index（AI） of CAL27 and CAL27-res increased significantly （CAL27： χ²=165.0797， P＜0.001； CAL27-res：χ²=196.5640， P＜0.001）.

Conclusion

EMT induced by TGF-β1 enhanced the cisplatin-resistance capacity in CAL27 and CAL27-res cells， accompanied by growth inhibition and apoptosis accelerated.

Key words: Tongue squamous cell carcinoma, Epithelial-mesenchymal transition, Transforming growth factor-β1, Drug resistance

Mo LIU, Bin ZHANG, Cheng WANG, Xi-qiang LIU, Jian-ning WANG, Hong-zhang HUANG. Study on epithelial-mesenchymal transition induced by TGF-β1 enhances cisplatin-resistance capacity of human tongue squamous cell carci[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2012, 6(03): 223-231.

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Figures/Tables 9

References 26

1	Franceschi D， Gupta R， Spiro RH， et al. Improved survival in the treatment of squamous carcinoma of the oral tongue. Am J Surg， 1993，166（4）：360-365.
2	Kajiyama H， Shibata K， Terauchi M， et al. Chemoresistance to paclitaxel induces epithelial-mesenchymal transition and enhances metastatic potential for epithelial ovarian carcinoma cells. Int J Oncol， 2007，31（2）：277-283.
3	Thomson S， Buck E， Petti F， et al. Epithelial to mesenchymal transition is a determinant of sensitivity of non-small-cell lung carcinoma cell lines and xenografts to epidermal growth factor receptor inhibition. Cancer Res， 2005，65（20）：9455-9462.
4	Yauch RL， Januario T， Eberhard DA， et al. Epithelial versus mesenchymal phenotype determines in vitro sensitivity and predicts clinical activity of erlotinib in lung cancer patients.Clin Cancer Res， 2005，11（24 Pt 1）：8686-8698.
5	Yang AD， Fan F， Camp ER， et al. Chronic oxaliplatin resistance induces epithelial-to-mesenchymal transition in colorectal cancer cell lines. Clin Cancer Res， 2006，12（14 Pt 1）：4147-4153.
6	Sabbah M， Emami S， Redeuilh G， et al. Molecular signature and therapeutic perspective of the epithelial-to-mesenchymal transitions in epithelial cancers. Drug Resist Updat， 2008，11（4-5）：123-151.
7	Wendt MK， Allington TM， Schiemann WP. Mechanisms of the epithelial-mesenchymal transition by TGF-beta. Future Oncol，2009，5（8）：1145-1168.
8	Yang AD， Fan F， Camp ER， et al. Chronic oxaliplatin resistance induces epithelial-to-mesenchymal transition in colorectal cancer cell lines. Clin Cancer Res， 2006，12（14 Pt 1）：4147-4153.
9	Kajiyama H， Shibata K， Terauchi M， et al. Chemoresistance to paclitaxel induces epithelial-mesenchymal transition and enhances metastatic potential for epithelial ovarian carcinoma cells. Int J Oncol， 2007，31（2）：277-283.
10	Hiscox S， Jiang WG， Obermeier K， et al. Tamoxifen resistance in MCF7 cells promotes EMT-like behaviour and involves modulation of beta-catenin phosphorylation. Int J Cancer， 2006，18（2）：290-301.
11	Güngör C， Zander H， Effenberger KE， et al. Notch signaling activated by replication stress-induced expression of midkine drives epithelial-mesenchymal transition and chemoresistance in pancreatic cancer. Cancer Res， 2011，71（14）：5009-5019.
12	Guttilla IK， Phoenix KN， Hong X， et al. Prolonged mammosphere culture of MCF-7 cells induces an EMT and repression of the estrogen receptor by microRNAs. Breast Cancer Res Treat， 2012，132（1）：75-85.
13	Hoshino H， Miyoshi N， Nagai K， et al. Epithelialmesenchymal transition with expression of SNAI1-induced chemoresistance in colorectal cancer. Biochem Biophys Res Commun， 2009，390（3）：1061-1065.
14	Su HY， Lai HC， Lin YW， et al. Epigenetic silencing of SFRP5 is related to malignant phenotype and chemoresistance of ovarian cancer through Wnt signaling pathway. Int J Cancer，2010，127（3）：555-567.
15	Miettinen PJ， Ebner R， Lopez AR， et al. TGF-beta induced transdifferentiation of mammary epithelial cells to mesenchymal cells： involvement of typeⅠreceptors. J Cell Biol， 1994，127（6 Pt 2）：2021-2036.
16	Yu C， Liu Y， Huang D， et al. TGF-β1 mediates epithelial to mesenchymal transition via the TGF-β/Smad pathway in squamous cell carcinoma of the head and neck. Oncol Rep，2011，25（6）：1581-1587.
17	Pirozzi G， Tirino V， Camerlingo R， et al. Epithelial to mesenchymal transition by TGF-β1 induction increases stemness characteristics in primary non small cell lung cancer cell line.PLoS One， 2011，6（6）：e21548.
18	Wendt MK， Smith JA， Schiemann WP. Transforming growth factor-β-induced epithelial-mesenchymal transition facilitates epidermal growth factor-dependent breast cancer progression.Oncogene， 2010，29（49）：6485-6498.
19	Richter P， Umbreit C， Franz M， et al. EGF/TGFβ1 costimulation of oral squamous cell carcinoma cells causes an epithelial-mesenchymal transition cell phenotype expressing laminin 332. J Oral Pathol Med， 2011，40（1）：46-54.
20	Azarova AM，Lyu YL，Lin CP，et al.Roles of DNA topoisomeraseⅡisozymes in chemotherapy and secondary malignancies. Proc Natl Acad Sci U S A， 2007，104（26）：11014-11019.
21	Kim SH， Bark H， Choi CH. Mercury induces multidrug resistance-associated protein gene through p38 mitogen-activated protein kinase. Toxicol Lett， 2005，155（1）：143-150.
22	Guo X， Ma N， Wang J， et al. Increased p38-MAPK is responsible for chemotherapy resistance in human gastric cancer cells. BMC Cancer， 2008，8：375.
23	Meyerzu Schwabedissen HE， Grube M， Dreisbach A， et al.Epidermal growth factor-mediated activation of the map kinase cascade results in altered expression and function of ABCG2（BCRP）. Drug Metab Dispos， 2006，34（4）：524-533.
24	Yoon H， Min JK， Lee JW， et al. Acquisition of chemoresistance in intrahepatic cholangiocarcinoma cells by activation of AKT and extracellular signal-regulated kinase （ERK）1/2. Biochem Biophys Res Commun， 2011，405（3）：333-337.
25	Nickl-Jockschat T， Arslan F， Doerfelt A， et al. An imbalance between Smad and MAPK pathways is responsible for TGF-beta tumor promoting effects in high-grade gliomas. Int J Oncol，2007，30（2）：499-507.
26	Song J. EMT or apoptosis： a decision for TGF-β. Cell Res，2007，17（4）：289-290.

项目	CAL27	CAL27/EMT	CAL27-res	CAL27-res/EMT
G0/G1	34.92±3.20	63.90±7.43	40.21±5.16	79.12±8.38
G2/M	7.29±2.01	6.37±2.87	19.58±3.02	11.55±3.14
S	57.79±6.74	29.73±2.23	40.21±5.87	9.33±3.11
PI	65.08±6.24	36.10±4.65^a	59.79±6.32	20.88±2.59^a
AI	0.53±0.21	2.89±0.24^a	1.97±0.17	5.83±1.04^a

项目	CAL27	CAL27/EMT	CAL27-res	CAL27-res/EMT
G0/G1	34.92±3.20	63.90±7.43	40.21±5.16	79.12±8.38
G2/M	7.29±2.01	6.37±2.87	19.58±3.02	11.55±3.14
S	57.79±6.74	29.73±2.23	40.21±5.87	9.33±3.11
PI	65.08±6.24	36.10±4.65^a	59.79±6.32	20.88±2.59^a
AI	0.53±0.21	2.89±0.24^a	1.97±0.17	5.83±1.04^a

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