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Biological Behavior of Carcinoma Associated Fibroblasts of Oral Carcinoma in Vitro

Author: LiQiang
Tutor: XingShuZhong
School: Nanjing Medical University
Course: Clinical Stomatology
Keywords: Oral cancer Cancer-associated fibroblasts Fibroblast activation protein Attack Transwell chamber
CLC: R739.8
Type: Master's thesis
Year: 2009
Downloads: 16
Quote: 0
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Abstract


Objective To investigate the in vitro proliferation of human oral gingival squamous cell carcinoma associated fibroblasts (carcinoma associated fibroblasts, CAFs), adhesion, invasion, migration, and study its special protein, normal oral gingival fibroblast cells (normal fibrolasts NFs) as a control; vitro study abnormal fibroblast activation protein (the activation protein, FAP), fibroblast invasion and migration ability of its cells, α-smooth muscle actin (α-smooth muscle actin, α-SMA) and matrix metalloproteinase Protease -2 (matrix metalloprotease-2, MMP-2) expression level of relationship. Methods Tissue blocks affixed to the same patient's mouth gums CAFs and of NFs wall method in primary culture; take three generations of purified cell morphological observation, immunohistochemical staining, cell proliferation activity, adherence rate of wound healing assay and transwell invasion test, compare The two cells in vitro biological behavior differences; applications Matrigel gel pack off the transwell chambers preliminary screening of non-invasive cell lines CAFs-NI (carcinoma associated fibroblasts-non-invasive) and invasive cell lines CAFs-HI (carcinoma associated fibroblasts hyper-invasive), immunohistochemical staining, detection Western Bloting immunoblotting flow cytometry immunofluorescence counting, analysis of its FAP, α-SMA, MMP-2 three kinds of protein expression level differences. Results 1 purified CAFs and NFs microscopy showed different cell morphology and growth characteristics; Both cells showed the typical 'S' shaped growth curve, but the CAFs vitro proliferation activity was significantly higher than that of NFs; each time point of the two types of cells the adherence rate is relatively the CAFs is also higher than that of NFs; the scratch experiments show, CAFs enhanced cell motility after 24 hours 0.8mm scratches basic repair of NFs relatively low migration transwell invasion ability of CAFs also significantly higher than the high NFs (P lt; 0.05). 2 immunohistochemical staining found that the two types of cells showed positive expression of vimentin (Vimentin) and negative expression of keratin (Cytokeratin), but CAFs can FAP, α-SMA and specific expression of MMP-2, while NFs was negative. 3 immunohistochemical staining, Western Bloting immune mark detection, flow cytometry immunofluorescence counting results show the FAP expression levels of α-SMA and MMP-2 three proteins in the highly invasive cell lines CAFs-HI than CAFs-NI high (P lt; 0.05). Conclusion the oral gingival CAFs vitro proliferation, adhesion, migration and invasion abilities were higher than those of NFs; oral gingival CAFs characteristic expression of FAP, α-SMA and MMP-2 related proteins; the oral gingival CAFs high vitro invasion ability their FAP, MMP-2, α-SMA protein expression levels positively correlated, these three proteins may mediate the CAFs some special biological behavior.

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CLC: > Medicine, health > Oncology > Oral cavity, maxillofacial tumors
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