臺灣博碩士論文加值系統

肺癌是目前國人死亡率最高的疾病之一，肺癌的轉移是導致病患死亡的主因，而肺癌的治療上又缺乏診斷轉移的生物指標，因此肺癌的轉移一直都是大家努力研究的目標。Desmosome (橋粒)是細胞與細胞黏附的主要結構之一，許多研究認為Desmosome在訊息傳遞亦扮演重要的角色。Desmocollin-2 (DSC2)是一種跨膜蛋白為Desmosome主要組成蛋白之一，分布於大部分的上皮細胞。近年來研究指出DSC2在大腸直腸癌、胃癌和食道癌都的研究證明DSC2表現減少與癌細胞的增生、轉移、侵入有關。然而DSC2基因與肺癌的關係性及在肺癌細胞中所扮演的角色尚未清楚。因此本研究探討DSC2是否會影響肺癌細胞的功能。首先利用即時定量PCR和西方墨點法分析DSC2在CL1-0、CL1-5和A549肺癌細胞株mRNA與蛋白質的表現量，結果顯示DSC2在CL1-0的表現量比CL1-5和A549高出許多。初步確定DSC2在不同肺癌細胞株表現會有差異。所以我們利用CL1-0細胞株藉由shRNA的方式抑制DSC2之後分析DSC2基因對細胞功能的影響。結果證明抑制DSC2之後會促進肺癌細胞增生、遷移和侵入能力，並且細胞會產生Epithelial to mesenchymal transition (EMT)現象。這表示DSC2在腫瘤發展中扮演一個重要的角色。為了瞭解DSC2是如何影響肺癌細胞增生、遷移和侵入能力，我們利用Microarray assay方法分析發現DSC2可能藉由調節：1. MMP10、SHISA3、NDRG1和SLIT2表現可能影響細胞轉移。2. Desmosome的其他組成蛋白JUP、PKP2、DSP、DSC3、DSC1和DSG2表現則影響細胞黏附及移動能力。3. EGFR和DLC1表現可能影響細胞生長。4. IL18和SOX4表現可能影響細胞凋亡。本研究針對DSC2在肺癌中的調控機制的探討對於未來肺癌病人的治療與管理是有很大的幫助。

Lung cancer is the most common cause of cancer death in the world, lung cancer patients died mainly due to metastasis, the lack of treatment of lung cancer diagnostic biomarkers transfer. Thus lung metastasis has been a major research goal. Desmosome are cell adhesion structures, many research now believe desmosome is the signal center. Desmocollin-2 (DSC2) is a transmembrane protein as the one of main component of the desmosomal proteins, major distribution in the epithelial cells. Recent studies indicate DSC2 in colorectal cancer, stomach cancer, oral cancer, esophageal cancer, and have proven DSC2 expression reducing cancer cell proliferation, metastasis, invasion. However, the role of the DSC2 gene in lung cancer cells is still unclear. Therefore, this study examines the DSC2 whether affect the development of lung cancer. Using real-time PCR and Western blot analysis DSC2 in CL1-0, CL1-5 and A549 lung cancer cell line mRNA and protein expression levels, the results indicate CL1-0 DSC2 expression than CL1-5 and A549 higher. Preliminary evidence in various cancer cell lines, DSC2 expression is different. Using CL1-0 cell line inhibition by shRNA after analysis of gene function DSC2. The results demonstrate inhibition DSC2 could promote cell proliferation, migration and invasion capabilities, and also lead to cell epithelial to mesenchymal transition (EMT) phenomenon. DSC2 expressed in tumor development plays an important role. In order to understand how DSC2 affects cell proliferation, migration and invasion capacity, we used microarray assay method of analysis found DSC2 possible by adjusting: 1. MMP10, SHISA3, NDRG1 and SLIT2 performance may affect cell metastasis. 2. Desmosome other components of the protein JUP, PKP2, DSP, DSC3, DSC1 DSG2 performance and impact of cell adhesion and movement. 3. EGFR and DLC1 expression may affect cell growth. 4. IL18 and SOX4 performance may affect apoptosis. This study further explored DSC2 regulatory mechanisms in lung cancer patients for future treatment and management is a great help.

封面內頁
簽名頁
中文摘要 iii
英文摘要 v
誌謝 vii
目錄 viii
圖目錄 xi
表目錄 xii

1. 前言 1
1.1 Desmosome的結構及組成 1
1.2 Desmosome與其他疾病和癌症的關係 2
1.3 Desmocollin-2 (DSC2)與癌症相關研究 3
1.4 肺癌 4
1.5 癌轉移 6
1.6 肺癌轉移相關研究 8
1.7 上皮細胞轉間葉細胞 (Epithelial–mesenchymal transition，EMT) 9
2. 研究動機 13
3. 實驗設計與流程 14
4. 材料與方法 15
4.1 細胞株 15
4.2 細胞繼代培養 15
4.3細胞凍存 16
4.4 細菌凍存 16
4.5 建立抑制DSC2之系統 17
4.6 質體DNA萃取 17
4.6.1 傳統法質體DNA萃取 17
4.6.2 萃取質體DNA 18
4.7 DNA電泳 19
4.8 DNA膠體純化 20
4.9 轉染 (Transfection) 20
4.10 連續稀釋挑Stable Clone 21
4.11 RNA萃取 22
4.12 cDNA合成反應 22
4.13 即時定量PCR (QPCR) 23
4.14 西方墨點法(Western blot) 24
4.14.1 SDS膠體的製備 24
4.14.2蛋白質樣品製備 25
4.14.3蛋白質的定量 26
4.14.4 SDS膠體電泳 26
4.14.5 半乾式電轉 (Semi-Dry Transfer) 27
4.14.6 抗體雜合 28
4.15 MTT assay 28
4.16 細胞群落分析 (Colony formation assay) 29
4.17 Soft agar assay 29
4.18 細胞遷移能力分析 (Wound healing assay) 30
4.19 細胞侵入能力分析 (Transwell invasion assay) 30
4.20 Micoarray分析 31
5. 結果 33
5.1 分析DSC2基因在CL1-0、CL1-5和A549肺癌細胞株中mRNA表現量的差異 33
5.2 分析DSC2基因在CL1-0、CL1-5和A549肺癌細胞株中 蛋白質表現量的差異 33
5.3 利用即時定量PCR方法確定抑制DSC2基因之肺癌模式 細胞的建立 34
5.4 利用西方墨點法確定抑制DSC2基因之肺癌模式細胞的 建立 35
5.5 抑制DSC2基因表現會促進肺癌細胞的增生能力 35
5.6 抑制DSC2基因表現會促進肺癌細胞群落形成的能力 36
5.7 抑制DSC2基因表現會促進肺癌細胞在Anchorage-independent環境下細胞株群落形成的能力 36
5.8 抑制DSC2基因表現會促進肺癌細胞的遷移能力 37
5.9 抑制DSC2基因表現會促進肺癌細胞侵入能力 37
5.10 抑制DSC2基因表現會促進肺癌細胞表現EMT型態 38
5.11 抑制DSC2基因表現可能影響的訊號路徑 39
5.12 在增生、細胞凋亡、細胞黏附、細胞週期和轉移這五個主要的細胞功能中受DSC2所影響的基因 39
5.13 抑制DSC2基因表現所促進的基因 40
5.14 Microarray assay data與即時定量PCR data 41
6. 討論 42
7. 結論 49
參考文獻 66
附錄 81

1.Chemotherapy in non-small cell lung cancer: a meta-analysis using updated data on individual patients from 52 randomised clinical trials. Non-small Cell Lung Cancer Collaborative Group. BMJ. 1995;311:899-909.
2.Aktary Z, Pasdar M. Plakoglobin: role in tumorigenesis and metastasis. Int J Cell Biol. 2012;2012:189521.
3.Alazawi WO, Morris LS, Stanley MA, Garrod DR, Coleman N. Altered expression of desmosomal components in high-grade squamous intraepithelial lesions of the cervix. Virchows Arch. 2003;443:51-6.
4.Alcalai R, Metzger S, Rosenheck S, Meiner V, Chajek-Shaul T. A recessive mutation in desmoplakin causes arrhythmogenic right ventricular dysplasia, skin disorder, and woolly hair. J Am Coll Cardiol. 2003;42:319-27.
5.Alroy J, Pauli BU, Weinstein RS. Correlation between numbers of desmosomes and the aggressiveness of transitional cell carcinoma in human urinary bladder. Cancer. 1981;47:104-12.
6.Anami K, Oue N, Noguchi T, Sakamoto N, Sentani K, Hayashi T, et al. Search for transmembrane protein in gastric cancer by the Escherichia coli ampicillin secretion trap: expression of DSC2 in gastric cancer with intestinal phenotype. J Pathol. 2010;221:275-84.
7.Armstrong DK, McKenna KE, Purkis PE, Green KJ, Eady RA, Leigh IM, et al. Haploinsufficiency of desmoplakin causes a striate subtype of palmoplantar keratoderma. Hum Mol Genet. 1999;8:143-8.
8.Bazzi H, Christiano AM. Broken hearts, woolly hair, and tattered skin: when desmosomal adhesion goes awry. Curr Opin Cell Biol. 2007;19:515-20.
9.Beug H. Breast cancer stem cells: eradication by differentiation therapy? Cell. 2009;138:623-5.
10.Bhat-Nakshatri P, Appaiah H, Ballas C, Pick-Franke P, Goulet R, Jr., Badve S, et al. SLUG/SNAI2 and tumor necrosis factor generate breast cells with CD44+/CD24- phenotype. BMC Cancer. 2010;10:411.
11.Biedermann K, Vogelsang H, Becker I, Plaschke S, Siewert JR, Hofler H, et al. Desmoglein 2 is expressed abnormally rather than mutated in familial and sporadic gastric cancer. J Pathol. 2005;207:199-206.
12.Biesalski HK, Bueno de Mesquita B, Chesson A, Chytil F, Grimble R, Hermus RJ, et al. European Consensus Statement on Lung Cancer: risk factors and prevention. Lung Cancer Panel. CA Cancer J Clin. 1998;48:167-76; discussion 4-6.
13.Birchmeier W, Behrens J. Cadherin expression in carcinomas: role in the formation of cell junctions and the prevention of invasiveness. Biochim Biophys Acta. 1994;1198:11-26.
14.Brabletz T, Jung A, Spaderna S, Hlubek F, Kirchner T. Opinion: migrating cancer stem cells - an integrated concept of malignant tumour progression. Nat Rev Cancer. 2005;5:744-9.
15.Bremnes RM, Veve R, Hirsch FR, Franklin WA. The E-cadherin cell-cell adhesion complex and lung cancer invasion, metastasis, and prognosis. Lung Cancer. 2002;36:115-24.
16.Brennan D, Hu Y, Joubeh S, Choi YW, Whitaker-Menezes D, O'Brien T, et al. Suprabasal Dsg2 expression in transgenic mouse skin confers a hyperproliferative and apoptosis-resistant phenotype to keratinocytes. J Cell Sci. 2007;120:758-71.
17.Brennan D, Mahoney MG. Increased expression of Dsg2 in malignant skin carcinomas: A tissue-microarray based study. Cell Adh Migr. 2009;3:148-54.
18.Breuninger S, Reidenbach S, Sauer CG, Strobel P, Pfitzenmaier J, Trojan L, et al. Desmosomal plakophilins in the prostate and prostatic adenocarcinomas: implications for diagnosis and tumor progression. Am J Pathol. 2010;176:2509-19.
19.Buchanan FG, Wang D, Bargiacchi F, DuBois RN. Prostaglandin E2 regulates cell migration via the intracellular activation of the epidermal growth factor receptor. J Biol Chem. 2003;278:35451-7.
20.Cao R, Farnebo J, Kurimoto M, Cao Y. Interleukin-18 acts as an angiogenesis and tumor suppressor. FASEB J. 1999;13:2195-202.
21.Chambers AF, MacDonald IC, Schmidt EE, Morris VL, Groom AC. Clinical targets for anti-metastasis therapy. Adv Cancer Res. 2000;79:91-121.
22.Charpentier E, Lavker RM, Acquista E, Cowin P. Plakoglobin suppresses epithelial proliferation and hair growth in vivo. J Cell Biol. 2000;149:503-20.
23.Chen YJ, Chang JT, Lee L, Wang HM, Liao CT, Chiu CC, et al. DSG3 is overexpressed in head neck cancer and is a potential molecular target for inhibition of oncogenesis. Oncogene. 2007;26:467-76.
24.Cheng X, Den Z, Koch PJ. Desmosomal cell adhesion in mammalian development. Eur J Cell Biol. 2005;84:215-23.
25.Cheng X, Koch PJ. In vivo function of desmosomes. J Dermatol. 2004;31:171-87.
26.Chidgey M, Dawson C. Desmosomes: a role in cancer? Br J Cancer. 2007;96:1783-7.
27.Chu YW, Yang PC, Yang SC, Shyu YC, Hendrix MJ, Wu R, et al. Selection of invasive and metastatic subpopulations from a human lung adenocarcinoma cell line. Am J Respir Cell Mol Biol. 1997;17:353-60.
28.Collins JE, Legan PK, Kenny TP, MacGarvie J, Holton JL, Garrod DR. Cloning and sequence analysis of desmosomal glycoproteins 2 and 3 (desmocollins): cadherin-like desmosomal adhesion molecules with heterogeneous cytoplasmic domains. J Cell Biol. 1991;113:381-91.
29.Cronier L, Crespin S, Strale PO, Defamie N, Mesnil M. Gap junctions and cancer: new functions for an old story. Antioxid Redox Signal. 2009;11:323-38.
30.Cui T, Chen Y, Yang L, Knosel T, Huber O, Pacyna-Gengelbach M, et al. The p53 target gene desmocollin 3 acts as a novel tumor suppressor through inhibiting EGFR/ERK pathway in human lung cancer. Carcinogenesis. 2012;33:2326-33.
31.Culkins CC, Setzer SV. Spotting desmosomes: the first 100 years. J Invest Dermatol. 2007;127:E2-3.
32.Curran S, Murray GI. Matrix metalloproteinases in tumour invasion and metastasis. J Pathol. 1999;189:300-8.
33.D'Amico TA, Massey M, Herndon JE, 2nd, Moore MB, Harpole DH, Jr. A biologic risk model for stage I lung cancer: immunohistochemical analysis of 408 patients with the use of ten molecular markers. J Thorac Cardiovasc Surg. 1999;117:736-43.
34.Dasari V, Gallup M, Lemjabbar H, Maltseva I, McNamara N. Epithelial-mesenchymal transition in lung cancer: is tobacco the "smoking gun"? Am J Respir Cell Mol Biol. 2006;35:3-9.
35.de Boer CJ, van Dorst E, van Krieken H, Jansen-van Rhijn CM, Warnaar SO, Fleuren GJ, et al. Changing roles of cadherins and catenins during progression of squamous intraepithelial lesions in the uterine cervix. Am J Pathol. 1999;155:505-15.
36.de Herreros AG, Peiro S, Nassour M, Savagner P. Snail family regulation and epithelial mesenchymal transitions in breast cancer progression. J Mammary Gland Biol Neoplasia. 2010;15:135-47.
37.Delva E, Tucker DK, Kowalczyk AP. The desmosome. Cold Spring Harb Perspect Biol. 2009;1:a002543.
38.Demirag GG, Sullu Y, Yucel I. Expression of Plakophilins (PKP1, PKP2, and PKP3) in breast cancers. Med Oncol. 2012;29:1518-22.
39.Derynck R, Akhurst RJ, Balmain A. TGF-beta signaling in tumor suppression and cancer progression. Nat Genet. 2001;29:117-29.
40.Dusek RL, Attardi LD. Desmosomes: new perpetrators in tumour suppression. Nat Rev Cancer. 2011;11:317-23.
41.Edme N, Downward J, Thiery JP, Boyer B. Ras induces NBT-II epithelial cell scattering through the coordinate activities of Rac and MAPK pathways. J Cell Sci. 2002;115:2591-601.
42.El-Rayes BF, LoRusso PM. Targeting the epidermal growth factor receptor. Br J Cancer. 2004;91:418-24.
43.Fang JY, Richardson BC. The MAPK signalling pathways and colorectal cancer. Lancet Oncol. 2005;6:322-7.
44.Fang WK, Gu W, Li EM, Wu ZY, Shen ZY, Shen JH, et al. Reduced membranous and ectopic cytoplasmic expression of DSC2 in esophageal squamous cell carcinoma: an independent prognostic factor. Hum Pathol. 2010;41:1456-65.
45.Funakoshi S, Ezaki T, Kong J, Guo RJ, Lynch JP. Repression of the desmocollin 2 gene expression in human colon cancer cells is relieved by the homeodomain transcription factors Cdx1 and Cdx2. Mol Cancer Res. 2008;6:1478-90.
46.Furukawa C, Daigo Y, Ishikawa N, Kato T, Ito T, Tsuchiya E, et al. Plakophilin 3 oncogene as prognostic marker and therapeutic target for lung cancer. Cancer Res. 2005;65:7102-10.
47.Garrod D, Chidgey M. Desmosome structure, composition and function. Biochim Biophys Acta. 2008;1778:572-87.
48.Gavert N, Ben-Ze'ev A. beta-Catenin signaling in biological control and cancer. J Cell Biochem. 2007;102:820-8.
49.Giannoni E, Bianchini F, Masieri L, Serni S, Torre E, Calorini L, et al. Reciprocal activation of prostate cancer cells and cancer-associated fibroblasts stimulates epithelial-mesenchymal transition and cancer stemness. Cancer Res. 2010;70:6945-56.
50.Giard DJ, Aaronson SA, Todaro GJ, Arnstein P, Kersey JH, Dosik H, et al. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst. 1973;51:1417-23.
51.Gill JH, Kirwan IG, Seargent JM, Martin SW, Tijani S, Anikin VA, et al. MMP-10 is overexpressed, proteolytically active, and a potential target for therapeutic intervention in human lung carcinomas. Neoplasia. 2004;6:777-85.
52.Goldstraw P. Editorial comment: Revisions to the 7th edition of TNM for lung cancer: data are good but prospective data are better! Eur J Cardiothorac Surg. 2012;42:811-2.
53.Grivaux M, Zureik M, Marsal L, Asselain B, Peureux M, Chavaillon JM, et al. Five-year survival for lung cancer patients managed in general hospitals. Rev Mal Respir. 2011;28:e31-8.
54.Gupta PB, Onder TT, Jiang G, Tao K, Kuperwasser C, Weinberg RA, et al. Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell. 2009;138:645-59.
55.Hakimelahi S, Parker HR, Gilchrist AJ, Barry M, Li Z, Bleackley RC, et al. Plakoglobin regulates the expression of the anti-apoptotic protein BCL-2. J Biol Chem. 2000;275:10905-11.
56.Harada T, Shinohara M, Nakamura S, Shimada M, Oka M. Immunohistochemical detection of desmosomes in oral squamous cell carcinomas: correlation with differentiation, mode of invasion, and metastatic potential. Int J Oral Maxillofac Surg. 1992;21:346-9.
57.Harrington KJ, Syrigos KN. The role of E-cadherin-catenin complex: more than an intercellular glue? Ann Surg Oncol. 2000;7:783-8.
58.Hecht SS, Hochalter JB, Villalta PW, Murphy SE. 2'-Hydroxylation of nicotine by cytochrome P450 2A6 and human liver microsomes: formation of a lung carcinogen precursor. Proc Natl Acad Sci U S A. 2000;97:12493-7.
59.Heid HW, Schmidt A, Zimbelmann R, Schafer S, Winter-Simanowski S, Stumpp S, et al. Cell type-specific desmosomal plaque proteins of the plakoglobin family: plakophilin 1 (band 6 protein). Differentiation. 1994;58:113-31.
60.Hiraki A, Shinohara M, Ikebe T, Nakamura S, Kurahara S, Garrod DR. Immunohistochemical staining of desmosomal components in oral squamous cell carcinomas and its association with tumour behaviour. Br J Cancer. 1996;73:1491-7.
61.Hiratsuka S, Nakamura K, Iwai S, Murakami M, Itoh T, Kijima H, et al. MMP9 induction by vascular endothelial growth factor receptor-1 is involved in lung-specific metastasis. Cancer Cell. 2002;2:289-300.
62.Huber O. Structure and function of desmosomal proteins and their role in development and disease. Cell Mol Life Sci. 2003;60:1872-90.
63.Hur W, Rhim H, Jung CK, Kim JD, Bae SH, Jang JW, et al. SOX4 overexpression regulates the p53-mediated apoptosis in hepatocellular carcinoma: clinical implication and functional analysis in vitro. Carcinogenesis. 2010;31:1298-307.
64.Jemal A, Thomas A, Murray T, Thun M. Cancer statistics, 2002. CA Cancer J Clin. 2002;52:23-47.
65.Jenndahl LE, Isakson P, Baeckstrom D. c-erbB2-induced epithelial-mesenchymal transition in mammary epithelial cells is suppressed by cell-cell contact and initiated prior to E-cadherin downregulation. Int J Oncol. 2005;27:439-48.
66.Justilien V, Regala RP, Tseng IC, Walsh MP, Batra J, Radisky ES, et al. Matrix metalloproteinase-10 is required for lung cancer stem cell maintenance, tumor initiation and metastatic potential. PLoS One. 2012;7:e35040.
67.Khan K, Hardy R, Haq A, Ogunbiyi O, Morton D, Chidgey M. Desmocollin switching in colorectal cancer. Br J Cancer. 2006;95:1367-70.
68.Kljuic A, Bazzi H, Sundberg JP, Martinez-Mir A, O'Shaughnessy R, Mahoney MG, et al. Desmoglein 4 in hair follicle differentiation and epidermal adhesion: evidence from inherited hypotrichosis and acquired pemphigus vulgaris. Cell. 2003;113:249-60.
69.Knosel T, Chen Y, Hotovy S, Settmacher U, Altendorf-Hofmann A, Petersen I. Loss of desmocollin 1-3 and homeobox genes PITX1 and CDX2 are associated with tumor progression and survival in colorectal carcinoma. Int J Colorectal Dis. 2012;27:1391-9.
70.Kogan G, Soltes L, Stern R, Gemeiner P. Hyaluronic acid: a natural biopolymer with a broad range of biomedical and industrial applications. Biotechnol Lett. 2007;29:17-25.
71.Kolegraff K, Nava P, Helms MN, Parkos CA, Nusrat A. Loss of desmocollin-2 confers a tumorigenic phenotype to colonic epithelial cells through activation of Akt/beta-catenin signaling. Mol Biol Cell. 2011;22:1121-34.
72.Kolligs FT, Kolligs B, Hajra KM, Hu G, Tani M, Cho KR, et al. gamma-catenin is regulated by the APC tumor suppressor and its oncogenic activity is distinct from that of beta-catenin. Genes Dev. 2000;14:1319-31.
73.Kottke MD, Delva E, Kowalczyk AP. The desmosome: cell science lessons from human diseases. J Cell Sci. 2006;119:797-806.
74.Kundu ST, Gosavi P, Khapare N, Patel R, Hosing AS, Maru GB, et al. Plakophilin3 downregulation leads to a decrease in cell adhesion and promotes metastasis. Int J Cancer. 2008;123:2303-14.
75.Kurzen H, Munzing I, Hartschuh W. Expression of desmosomal proteins in squamous cell carcinomas of the skin. J Cutan Pathol. 2003;30:621-30.
76.Liao YC, Lo SH. Deleted in liver cancer-1 (DLC-1): a tumor suppressor not just for liver. Int J Biochem Cell Biol. 2008;40:843-7.
77.Lorch JH, Klessner J, Park JK, Getsios S, Wu YL, Stack MS, et al. Epidermal growth factor receptor inhibition promotes desmosome assembly and strengthens intercellular adhesion in squamous cell carcinoma cells. J Biol Chem. 2004;279:37191-200.
78.Lustig B, Behrens J. The Wnt signaling pathway and its role in tumor development. J Cancer Res Clin Oncol. 2003;129:199-221.
79.MacDonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components, mechanisms, and diseases. Dev Cell. 2009;17:9-26.
80.Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 2008;133:704-15.
81.Markowitz SD, Bertagnolli MM. Molecular origins of cancer: Molecular basis of colorectal cancer. N Engl J Med. 2009;361:2449-60.
82.Matsugaki T, Zenmyo M, Hiraoka K, Fukushima N, Shoda T, Komiya S, et al. N-myc downstream-regulated gene 1/Cap43 expression promotes cell differentiation of human osteosarcoma cells. Oncol Rep. 2010;24:721-5.
83.Mazieres J, He B, You L, Xu Z, Jablons DM. Wnt signaling in lung cancer. Cancer Lett. 2005;222:1-10.
84.McLean GW, Carragher NO, Avizienyte E, Evans J, Brunton VG, Frame MC. The role of focal-adhesion kinase in cancer - a new therapeutic opportunity. Nat Rev Cancer. 2005;5:505-15.
85.Medina PP, Castillo SD, Blanco S, Sanz-Garcia M, Largo C, Alvarez S, et al. The SRY-HMG box gene, SOX4, is a target of gene amplification at chromosome 6p in lung cancer. Hum Mol Genet. 2009;18:1343-52.
86.Neal DE, Marsh C, Bennett MK, Abel PD, Hall RR, Sainsbury JR, et al. Epidermal-growth-factor receptors in human bladder cancer: comparison of invasive and superficial tumours. Lancet. 1985;1:366-8.
87.Neuber S, Muhmer M, Wratten D, Koch PJ, Moll R, Schmidt A. The desmosomal plaque proteins of the plakophilin family. Dermatol Res Pract. 2010;2010:101452.
88.Nolan DJ, Ciarrocchi A, Mellick AS, Jaggi JS, Bambino K, Gupta S, et al. Bone marrow-derived endothelial progenitor cells are a major determinant of nascent tumor neovascularization. Genes Dev. 2007;21:1546-58.
89.Nollet F, Kools P, van Roy F. Phylogenetic analysis of the cadherin superfamily allows identification of six major subfamilies besides several solitary members. J Mol Biol. 2000;299:551-72.
90.O'Keefe EJ, Erickson HP, Bennett V. Desmoplakin I and desmoplakin II. Purification and characterization. J Biol Chem. 1989;264:8310-8.
91.Onder TT, Gupta PB, Mani SA, Yang J, Lander ES, Weinberg RA. Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 2008;68:3645-54.
92.Oshiro MM, Kim CJ, Wozniak RJ, Junk DJ, Munoz-Rodriguez JL, Burr JA, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 2005;7:R669-80.
93.Oving IM, Clevers HC. Molecular causes of colon cancer. Eur J Clin Invest. 2002;32:448-57.
94.Pantel K, Passlick B, Vogt J, Stosiek P, Angstwurm M, Seen-Hibler R, et al. Reduced expression of plakoglobin indicates an unfavorable prognosis in subsets of patients with non-small-cell lung cancer. J Clin Oncol. 1998;16:1407-13.
95.Parker HR, Li Z, Sheinin H, Lauzon G, Pasdar M. Plakoglobin induces desmosome formation and epidermoid phenotype in N-cadherin-expressing squamous carcinoma cells deficient in plakoglobin and E-cadherin. Cell Motil Cytoskeleton. 1998;40:87-100.
96.Parkin DM, Bray FI, Devesa SS. Cancer burden in the year 2000. The global picture. Eur J Cancer. 2001;37 Suppl 8:S4-66.
97.Pedersen H, Brunner N, Francis D, Osterlind K, Ronne E, Hansen HH, et al. Prognostic impact of urokinase, urokinase receptor, and type 1 plasminogen activator inhibitor in squamous and large cell lung cancer tissue. Cancer Res. 1994;54:4671-5.
98.Peifer M, Polakis P. Wnt signaling in oncogenesis and embryogenesis--a look outside the nucleus. Science. 2000;287:1606-9.
99.Peinado H, Olmeda D, Cano A. Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer. 2007;7:415-28. migration. Nat Rev Mol Cell Biol. 2009;10:538-49.
103.Pinto D, Clevers H. Wnt control of stem cells and differentiation in the intestinal epithelium. Exp Cell Res. 2005;306:357-63.
104.Pittella F, Katsube K, Takemura T, Hashimoto T, Kawano T, Garrod D, et al. Perinuclear and cytoplasmic distribution of desmoglein in esophageal squamous cell carcinomas. Pathol Res Pract. 2001;197:85-91.
105.Rao TP, Kuhl M. An updated overview on Wnt signaling pathways: a prelude for more. Circ Res. 2010;106:1798-806.
106.Rappold I, Iwabuchi K, Date T, Chen J. Tumor suppressor p53 binding protein 1 (53BP1) is involved in DNA damage-signaling pathways. J Cell Biol. 2001;153:613-20.
107.Rickman L, Simrak D, Stevens HP, Hunt DM, King IA, Bryant SP, et al. N-terminal deletion in a desmosomal cadherin causes the autosomal dominant skin disease striate palmoplantar keratoderma. Hum Mol Genet. 1999;8:971-6.
108.Rosti G, Bevilacqua G, Bidoli P, Portalone L, Santo A, Genestreti G. Small cell lung cancer. Ann Oncol. 2006;17 Suppl 2:ii5-10.
109.Salden M, Splinter TA, Peters HA, Look MP, Timmermans M, van Meerbeeck JP, et al. The urokinase-type plasminogen activator system in resected non-small-cell lung cancer. Rotterdam Oncology Thoracic Study Group. Ann Oncol. 2000;11:327-32.
110.Savagner P. Leaving the neighborhood: molecular mechanisms involved during epithelial-mesenchymal transition. Bioessays. 2001;23:912-23.
111.Schaffer J. Vorlesungen über Histologie und Histogenese nebst Bemerkungen über Histotechnik und das Mikroscop: Engelmann; 1920.
112.Schmidt A, Koch PJ. Desmosomes: just cell adhesion or is there more? Cell Adh Migr. 2007;1:28-32.
113.Shih JY, Yang PC. The EMT regulator slug and lung carcinogenesis. Carcinogenesis. 2011;32:1299-304.
100.Perlikos F, Harrington KJ, Syrigos KN. Key molecular mechanisms in lung cancer invasion and metastasis: A comprehensive review. Crit Rev Oncol Hematol. 2013.
101.Perlikos F, Harrington KJ, Syrigos KN. Key molecular mechanisms in lung cancer invasion and metastasis: A comprehensive review. Crit Rev Oncol Hematol. 2013;87:1-11.
102.Petrie RJ, Doyle AD, Yamada KM. Random versus directionally persistent cell
114.Shinohara M, Hiraki A, Ikebe T, Nakamura S, Kurahara S, Shirasuna K, et al. Immunohistochemical study of desmosomes in oral squamous cell carcinoma: correlation with cytokeratin and E-cadherin staining, and with tumour behaviour. J Pathol. 1998;184:369-81.
115.Simcha I, Geiger B, Yehuda-Levenberg S, Salomon D, Ben-Ze'ev A. Suppression of tumorigenicity by plakoglobin: an augmenting effect of N-cadherin. J Cell Biol. 1996;133:199-209.
116.Singh A, Settleman J. EMT, cancer stem cells and drug resistance: an emerging axis of evil in the war on cancer. Oncogene. 2010;29:4741-51.
117.Strand TE, Rostad H, Damhuis RA, Norstein J. Risk factors for 30-day mortality after resection of lung cancer and prediction of their magnitude. Thorax. 2007;62:991-7.
118.Tang J, Wang J, Kong X, Yang J, Guo L, Zheng F, et al. Vascular endothelial growth factor promotes cardiac stem cell migration via the PI3K/Akt pathway. Exp Cell Res. 2009;315:3521-31.
119.Thiery JP. Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002;2:442-54.
120.Thiery JP, Acloque H, Huang RY, Nieto MA. Epithelial-mesenchymal transitions in development and disease. Cell. 2009;139:871-90.
121.Travis WD, World Health Organization., International Agency for Research on Cancer., International Association for the Study of Lung Cancer., International Academy of Pathology. Pathology and genetics of tumours of the lung, pleura, thymus and heart. Lyon Oxford: IARC Press Oxford University Press (distributor); 2004.
122.Tse BW, Russell PJ, Lochner M, Forster I, Power CA. IL-18 inhibits growth of murine orthotopic prostate carcinomas via both adaptive and innate immune mechanisms. PLoS One. 2011;6:e24241.
123.Tselepis C, Chidgey M, North A, Garrod D. Desmosomal adhesion inhibits invasive behavior. Proc Natl Acad Sci U S A. 1998;95:8064-9.
124.Tseng RC, Lee SH, Hsu HS, Chen BH, Tsai WC, Tzao C, et al. SLIT2 attenuation during lung cancer progression deregulates beta-catenin and E-cadherin and associates with poor prognosis. Cancer Res. 2010;70:543-51.
125.Ureshino H, Murakami Y, Watari K, Izumi H, Kawahara A, Kage M, et al. N-myc downstream regulated gene 1 (NDRG1) promotes metastasis of human scirrhous gastric cancer cells through epithelial mesenchymal transition. PLoS One. 2012;7:e41312.
126.Verkaar F, Zaman GJ. New avenues to target Wnt/beta-catenin signaling. Drug Discov Today. 2011;16:35-41.
127.Vilela MJ, Hashimoto T, Nishikawa T, North AJ, Garrod D. A simple epithelial cell line (MDCK) shows heterogeneity of desmoglein isoforms, one resembling pemphigus vulgaris antigen. J Cell Sci. 1995;108 ( Pt 4):1743-50.
128.Wagner EF, Nebreda AR. Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 2009;9:537-49.
129.Wakefield LM, Roberts AB. TGF-beta signaling: positive and negative effects on tumorigenesis. Curr Opin Genet Dev. 2002;12:22-9.
130.Walker-Daniels J, Coffman K, Azimi M, Rhim JS, Bostwick DG, Snyder P, et al. Overexpression of the EphA2 tyrosine kinase in prostate cancer. Prostate. 1999;41:275-80.
131.Whittock NV, Ashton GH, Dopping-Hepenstal PJ, Gratian MJ, Keane FM, Eady RA, et al. Striate palmoplantar keratoderma resulting from desmoplakin haploinsufficiency. J Invest Dermatol. 1999;113:940-6.
132.Winn RA, Bremnes RM, Bemis L, Franklin WA, Miller YE, Cool C, et al. gamma-Catenin expression is reduced or absent in a subset of human lung cancers and re-expression inhibits transformed cell growth. Oncogene. 2002;21:7497-506.
133.Woodburn JR. The epidermal growth factor receptor and its inhibition in cancer therapy. Pharmacol Ther. 1999;82:241-50.
134.Woodhouse EC, Chuaqui RF, Liotta LA. General mechanisms of metastasis. Cancer. 1997;80:1529-37.
135.Wu DL, Sui FY, Jiang XM, Jiang XH. Methylation in esophageal carcinogenesis. World J Gastroenterol. 2006;12:6933-40.
136.Wu GS. The functional interactions between the p53 and MAPK signaling pathways. Cancer Biol Ther. 2004;3:156-61.
137.Yang J, Weinberg RA. Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis. Dev Cell. 2008;14:818-29.
138.Yang L, Chen Y, Cui T, Knosel T, Zhang Q, Albring KF, et al. Desmoplakin acts as a tumor suppressor by inhibition of the Wnt/beta-catenin signaling pathway in human lung cancer. Carcinogenesis. 2012;33:1863-70.
139.Yang MH, Wu MZ, Chiou SH, Chen PM, Chang SY, Liu CJ, et al. Direct regulation of TWIST by HIF-1alpha promotes metastasis. Nat Cell Biol. 2008;10:295-305.
140.Yashiro M, Nishioka N, Hirakawa K. Decreased expression of the adhesion molecule desmoglein-2 is associated with diffuse-type gastric carcinoma. Eur J Cancer. 2006;42:2397-403.
141.Yoon SO, Park SJ, Yun CH, Chung AS. Roles of matrix metalloproteinases in tumor metastasis and angiogenesis. J Biochem Mol Biol. 2003;36:128-37.
142.Yuan BZ, Miller MJ, Keck CL, Zimonjic DB, Thorgeirsson SS, Popescu NC. Cloning, characterization, and chromosomal localization of a gene frequently deleted in human liver cancer (DLC-1) homologous to rat RhoGAP. Cancer Res. 1998;58:2196-9.
143.Zhang Q, Yin H, Liu P, Zhang H, She M. Essential role of HDL on endothelial progenitor cell proliferation with PI3K/Akt/cyclin D1 as the signal pathway. Exp Biol Med (Maywood). 2010;235:1082-92.
144.Zhang X, Zhu S, Luo G, Zheng L, Wei J, Zhu J, et al. Expression of MMP-10 in lung cancer. Anticancer Res. 2007;27:2791-5.
145.Zheng H, Fu G, Dai T, Huang H. Migration of endothelial progenitor cells mediated by stromal cell-derived factor-1alpha/CXCR4 via PI3K/Akt/eNOS signal transduction pathway. J Cardiovasc Pharmacol. 2007;50:274-80.