Premium
Esophageal squamous cell carcinoma model of 3D organotypic tissue culture system with genetic modification of epithelial growth factor receptor and p120‐catenin (1119.3)
Author(s) -
Yang Xuebin,
Lehman Heather,
Bruggeman Richard,
Nie Xingcao,
Stairs Douglas
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1119.3
Subject(s) - immunohistochemistry , keratin , epidermal growth factor receptor , epithelium , biology , pathology , catenin , esophagus , gene knockdown , receptor , cancer research , medicine , cell culture , signal transduction , microbiology and biotechnology , anatomy , wnt signaling pathway , genetics
Objectives : To evaluate whether our genetically (EGFR & p120ctn) modified 3D OTC esophageal epithelium resembles human ESCC. Methods: Four sets of OTC (5 per set) were obtained: control, p120ctn knockdown (p120ctn D ), EGFR upregulation (EGFR U ), and combined p120ctn D and EGFR U (p120ctn D &EGFR U ). Morphology was analyzed by H&E using ImageJ. Proliferation was quantified by Ki‐67 immunohistochemistry (IHC). Keratins (K) were studied by gene microarray and IHC in OTC, 10 human normal esophageal and 24 ESCC specimens. Results: In our OTC, p120ctn D showed mild disorganization, mildly increased cellularity and significantly increased proliferation (20±4% vs 31±4%, p<0.05) compared to the control. EGFR U showed moderate disorganization, marked increased cellularity (104±31 vs 192±64, p<0.05) and proliferation (32±7%, p<0.05). Interestingly, p120ctn D &EGFR U contained apparently transformed epithelium resulting in cellular invasion into the matrix. It had significantly increased cellularity (292±72), nuclear size (13.2±0.1 µm vs 11.1±0.8 µm) and proliferation (45±9%) (each p<0.05). p120ctn D &EGFR U had decreased RNA expression of K1, K4, K10 and K15. IHC studies showed similar keratin patterns between normal human esophagus and OTC control and between human ESCC and p120ctn D &EGFR U . Conclusions: Our genetically modified OTC esophageal epithelium resembles human ESCC with similar morphologic changes and keratin patterns.