MiR-200a inhibits epithelial-mesenchymal transition of pancreatic cancer stem cell

BMC Cancer

Zhiwei Wang¹  Mingyan Zhu¹  Yan Huang¹  Lei Wang¹  Qingsong Guo¹  Yao Wang¹  Shajun Zhu¹  Xiangjun Fan¹  Hui Zhu³  Xiaohong Li³  Jingjing Lu³  Yuhua Lu² 

[1]Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province 226001, P. R. China

[2]Visitor scholar of Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, NC 27101, USA

[3]Surgical comprehensive laboratory, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province 226001, P. R. China

关键词: miR-200a;    Pancreatic cancer;    EMT;    CSC;   

Others  :  859072
DOI  :  10.1186/1471-2407-14-85

 received in 2013-08-12, accepted in 2014-01-27,  发布年份 2014

【 摘 要 】

Background

Pancreatic cancer is one of the most aggressive cancers, and the aggressiveness of pancreatic cancer is in part due to its intrinsic and extrinsic drug resistance characteristics, which are also associated with the acquisition of epithelial-to-mesenchymal transition (EMT). Increasing evidence suggests that EMT-type cells share many biological characteristics with cancer stem-like cells. And miR-200 has been identified as a powerful regulator of EMT.

Methods

Cancer Stem Cells (CSCs) of human pancreatic cancer cell line PANC-1 were processed for CD24, CD44 and ESA multi-colorstaining, and sorted out on a BD FACS Aria II machine. RT-qPCR was performed using the miScript PCR Kit to assay the expression of miR-200 family. In order to find the role of miR-200a in the process of EMT, miR-200a mimic was transfected to CSCs.

Results

Pancreatic cancer cells with EMT phenotype displayed stem-like cell features characterized by the expression of cell surface markers CD24, CD44 and epithelial-specific antigen (ESA), which was associated with decreased expression of miR-200a. Moreover, overexpression of miR-200a was resulted in down-regulation of N-cadherin, ZEB1 and vimentin, but up-regulation of E-cadherin. In addition, miR-200a overexpression inhibited cell migration and invasion in CSCs.

Conclusion

In our study, we found that miR-200a played an important role in linking the characteristics of cancer stem-like cells with EMT-like cell signatures in pancreatic cancer. Selective elimination of cancer stem-like cells by reversing the EMT phenotype to mesenchymal-to-epithelial transition (MET) phenotype using novel agents would be useful for prevention and/or treatment of pancreatic cancer.

【 授权许可】

   
2014 Lu et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Xie CG, Wei SM, Chen JM, Xu XF, Cai JT, Chen QY, Jia LT: Down-regulation of GEP100 causes increase in E-cadherin levels and inhibits pancreatic cancer cell invasion. PLoS One 7(5):e37854.
• [2]Kallifatidis G, Rausch V, Baumann B, Apel A, Beckermann BM, Groth A, Mattern J, Li Z, Kolb A, Moldenhauer G, et al.: Sulforaphane targets pancreatic tumour-initiating cells by NF-kappaB-induced antiapoptotic signalling. Gut 2009, 58(7):949-963.
• [3]Bao B, Ali S, Kong D, Sarkar SH, Wang Z, Banerjee S, Aboukameel A, Padhye S, Philip PA, Sarkar FH: Anti-tumor activity of a novel compound-CDF is mediated by regulating miR-21, miR-200, and PTEN in pancreatic cancer. PLoS One 6(3):e17850.
• [4]Hermann PC, Bhaskar S, Cioffi M, Heeschen C: Cancer stem cells in solid tumors. Semin Cancer Biol 20(2):77-84.
• [5]Shackleton M: Normal stem cells and cancer stem cells: similar and different. Semin Cancer Biol 20(2):85-92.
• [6]Wen J, Park JY, Park KH, Chung HW, Bang S, Park SW, Song SY: Oct4 and Nanog expression is associated with early stages of pancreatic carcinogenesis. Pancreas 39(5):622-626.
• [7]Li Y, VandenBoom TG 2nd, Kong D, Wang Z, Ali S, Philip PA, Sarkar FH: Up-regulation of miR-200 and let-7 by natural agents leads to the reversal of epithelial-to-mesenchymal transition in gemcitabine-resistant pancreatic cancer cells. Cancer Res 2009, 69(16):6704-6712.
• [8]Kurahara H, Takao S, Maemura K, Mataki Y, Kuwahata T, Maeda K, Ding Q, Sakoda M, Iino S, Ishigami S, et al.: Epithelial-mesenchymal transition and mesenchymal-epithelial transition via regulation of ZEB-1 and ZEB-2 expression in pancreatic cancer. J Surg Oncol 105(7):655-661.
• [9]Watanabe S, Ueda Y, Akaboshi S, Hino Y, Sekita Y, Nakao M: HMGA2 maintains oncogenic RAS-induced epithelial-mesenchymal transition in human pancreatic cancer cells. Am J Pathol 2009, 174(3):854-868.
• [10]Peinado H, Portillo F, Cano A: Transcriptional regulation of cadherins during development and carcinogenesis. Int J Dev Biol 2004, 48(5–6):365-375.
• [11]Santisteban M, Reiman JM, Asiedu MK, Behrens MD, Nassar A, Kalli KR, Haluska P, Ingle JN, Hartmann LC, Manjili MH, et al.: Immune-induced epithelial to mesenchymal transition in vivo generates breast cancer stem cells. Cancer Res 2009, 69(7):2887-2895.
• [12]Kasper S: Identification, characterization, and biological relevance of prostate cancer stem cells from clinical specimens. Urol Oncol 2009, 27(3):301-303.
• [13]Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M, et al.: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 2008, 133(4):704-715.
• [14]Korpal M, Lee ES, Hu G, Kang Y: The miR-200 family inhibits epithelial-mesenchymal transition and cancer cell migration by direct targeting of E-cadherin transcriptional repressors ZEB1 and ZEB2. J Biol Chem 2008, 283(22):14910-14914.
• [15]Bullock MD, Sayan AE, Packham GK, Mirnezami AH: MicroRNAs: critical regulators of epithelial to mesenchymal (EMT) and mesenchymal to epithelial transition (MET) in cancer progression. Biol Cell 104(1):3-12.
• [16]Mongroo PS, Rustgi AK: The role of the miR-200 family in epithelial-mesenchymal transition. Cancer Biol Ther 10(3):219-222.
• [17]Hu X, Macdonald DM, Huettner PC, Feng Z, el Naqa IM, Schwarz JK, Mutch DG, Grigsby PW, Powell SN, Wang X: A miR-200 microRNA cluster as prognostic marker in advanced ovarian cancer. Gynecol Oncol 2009, 114(3):457-464.
• [18]Paterson EL, Kolesnikoff N, Gregory PA, Bert AG, Khew-Goodall Y, Goodall GJ: The microRNA-200 family regulates epithelial to mesenchymal transition. Sci World J 2008, 8:901-904.
• [19]Schliekelman MJ, Gibbons DL, Faca VM, Creighton CJ, Rizvi ZH, Zhang Q, Wong CH, Wang H, Ungewiss C, Ahn YH, et al.: Targets of the tumor suppressor miR-200 in regulation of the epithelial-mesenchymal transition in cancer. Cancer Res 71(24):7670-7682.
• [20]Soubani O, Ali AS, Logna F, Ali S, Philip PA, Sarkar FH: Re-expression of miR-200 by novel approaches regulates the expression of PTEN and MT1-MMP in pancreatic cancer. Carcinogenesis 33(8):1563-1571.
• [21]Chiou SH, Wang ML, Chou YT, Chen CJ, Hong CF, Hsieh WJ, Chang HT, Chen YS, Lin TW, Hsu HS, et al.: Coexpression of Oct4 and Nanog enhances malignancy in lung adenocarcinoma by inducing cancer stem cell-like properties and epithelial-mesenchymal transdifferentiation. Cancer Res 70(24):10433-10444.
• [22]Kong D, Li Y, Wang Z, Banerjee S, Ahmad A, Kim HR, Sarkar FH: miR-200 regulates PDGF-D-mediated epithelial-mesenchymal transition, adhesion, and invasion of prostate cancer cells. Stem Cells 2009, 27(8):1712-1721.
• [23]Polytarchou C, Iliopoulos D, Struhl K: An integrated transcriptional regulatory circuit that reinforces the breast cancer stem cell state. Proc Natl Acad Sci U S A 109(36):14470-14475.
• [24]Wang G, Guo X, Hong W, Liu Q, Wei T, Lu C, Gao L, Ye D, Zhou Y, Chen J, et al.: Critical regulation of miR-200/ZEB2 pathway in Oct4/Sox2-induced mesenchymal-to-epithelial transition and induced pluripotent stem cell generation. Proc Natl Acad Sci U S A 110(8):2858-2863.
• [25]Shankar S, Nall D, Tang SN, Meeker D, Passarini J, Sharma J, Srivastava RK: Resveratrol inhibits pancreatic cancer stem cell characteristics in human and KrasG12D transgenic mice by inhibiting pluripotency maintaining factors and epithelial-mesenchymal transition. PLoS One 6(1):e16530.
• [26]Ischenko I, Seeliger H, Kleespies A, Angele MK, Eichhorn ME, Jauch KW, Bruns CJ: Pancreatic cancer stem cells: new understanding of tumorigenesis, clinical implications. Langenbecks Arch Surg 395(1):1-10.
• [27]Wei HJ, Yin T, Zhu Z, Shi PF, Tian Y, Wang CY: Expression of CD44, CD24 and ESA in pancreatic adenocarcinoma cell lines varies with local microenvironment. Hepatobiliary Pancreat Dis Int 10(4):428-434.
• [28]Gach PC, Attayek PJ, Herrera G, Yeh JJ, Allbritton NL: Isolation and in vitro culture of rare cancer stem cells from patient-derived xenografts of pancreatic ductal adenocarcinoma. Anal Chem 2013, 85(15):7271-7278.
• [29]Fredebohm J, Boettcher M, Eisen C, Gaida MM, Heller A, Keleg S, Tost J, Greulich-Bode KM, Hotz-Wagenblatt A, Lathrop M, et al.: Establishment and characterization of a highly tumourigenic and cancer stem cell enriched pancreatic cancer cell line as a well defined model system. PLoS One 7(11):e48503.
• [30]Huang P, Wang CY, Gou SM, Wu HS, Liu T, Xiong JX: Isolation and biological analysis of tumor stem cells from pancreatic adenocarcinoma. World J Gastroenterol 2008, 14(24):3903-3907.
• [31]Ben-Ze’ev A: The use of two-dimensional gel electrophoresis in studies on the role of cytoskeletal plaque proteins as tumor suppressors. Electrophoresis 1996, 17(11):1752-1763.
• [32]Gou S, Liu T, Wang C, Yin T, Li K, Yang M, Zhou J: Establishment of clonal colony-forming assay for propagation of pancreatic cancer cells with stem cell properties. Pancreas 2007, 34(4):429-435.
• [33]Musch T, Oz Y, Lyko F, Breiling A: Nucleoside drugs induce cellular differentiation by caspase-dependent degradation of stem cell factors. PLoS One 5(5):e10726.
• [34]Chiou SH, Yu CC, Huang CY, Lin SC, Liu CJ, Tsai TH, Chou SH, Chien CS, Ku HH, Lo JF: Positive correlations of Oct-4 and Nanog in oral cancer stem-like cells and high-grade oral squamous cell carcinoma. Clin Cancer Res 2008, 14(13):4085-4095.
• [35]Luo W, Li S, Peng B, Ye Y, Deng X, Yao K: Embryonic stem cells markers SOX2, OCT4 and Nanog expression and their correlations with epithelial-mesenchymal transition in nasopharyngeal carcinoma. PLoS One 8(2):e56324.
• [36]Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y, Goodall GJ: The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol 2008, 10(5):593-601.
• [37]Sarkar FH, Li Y, Wang Z, Kong D: Pancreatic cancer stem cells and EMT in drug resistance and metastasis. Minerva Chir 2009, 64(5):489-500.
• [38]Gavert N, Ben-Ze’ev A: Epithelial-mesenchymal transition and the invasive potential of tumors. Trends Mol Med 2008, 14(5):199-209.
• [39]Cufi S, Vazquez-Martin A, Oliveras-Ferraros C, Martin-Castillo B, Joven J, Menendez JA: Metformin against TGFbeta-induced epithelial-to-mesenchymal transition (EMT): from cancer stem cells to aging-associated fibrosis. Cell Cycle 9(22):4461-4468.
• [40]Wu Y, Zhou BP: New insights of epithelial-mesenchymal transition in cancer metastasis. Acta Biochim Biophys Sin (Shanghai) 2008, 40(7):643-650.
• [41]Zavadil J, Bottinger EP: TGF-beta and epithelial-to-mesenchymal transitions. Oncogene 2005, 24(37):5764-5774.
• [42]Leong KG, Niessen K, Kulic I, Raouf A, Eaves C, Pollet I, Karsan A: Jagged1-mediated Notch activation induces epithelial-to-mesenchymal transition through Slug-induced repression of E-cadherin. J Exp Med 2007, 204(12):2935-2948.
• [43]Pirozzi G, Tirino V, Camerlingo R, Franco R, la Rocca A, Liguori E, Martucci N, Paino F, Normanno N, Rocco G: Epithelial to mesenchymal transition by TGFbeta-1 induction increases stemness characteristics in primary non small cell lung cancer cell line. PLoS One 6(6):e21548.