- 學位論文
TIMP-1和MMPs活性對翼狀贅片細胞侵犯能力的影響
Effect of MMPs and TIMP-1 activity in pterygium invation
摘要
翼狀贅片(pterygium) 是一種眼睛表面常見的疾病，它是一種富含纖維血管的組織，會從結膜往角膜處增生及侵犯，目前被認為可能是ㄧ種腫瘤。已知腫瘤細胞的轉移和侵犯能力與基質金屬蛋白酶 (Matrix metalloproteinases; MMPs)與 基質金屬蛋白酶抑制蛋白 (tissue inhibitors tissue inhibitors of MMPs; TIMPs)的交互作用有關。MMP的分類是依其分解的受質的種類而定，而從被侵犯的角膜組織膠原組成來看，MMP-9與MMP-10可能參與翼狀贅片的侵犯作用，另外，MMPs受到其抑制蛋白 TIMP 調控。本研究利用化學免疫染色法分析MMP-9、MMP-10、與TIMP在翼狀贅片組織的表現狀況 。實驗收集82個翼狀贅片檢體及30個正常的結膜組織。 發覺在82個翼狀贅片的檢體中， MMP-9陽性表現者有29位佔35.4%，具MMP-10陽性表現者有28位佔34.1%， TIMP-1陽性表現者有59位佔72.0%。MMPs的染色表現局限在表皮細胞的細胞質中。而TIMP-1則可在翼狀贅片的表皮細胞、纖維母細胞和角膜上皮細胞偵測到。 為了進一步評估翼狀贅片侵犯能力的表現是否受TIMP-1所影響，本研究在培養的初代翼狀贅片細胞中，以siRNA剔除TIMP基因表現， 結果顯示剔除TIMP-1表現的細胞相較於對照組其侵犯和轉移能力有增加的現象。因此推測 MMP-9和 MMP-10在翼狀贅片的侵犯作用與腫瘤特性扮演一定的角色，而TIMPs的表現則可能對於翼狀贅片的侵犯能力有抑制作用。
並列摘要
Pterygium is a common ocular surface disease. It is a fibrovascular tissue which originate near the corneal-conjunctival junction and centripetally to encroach on the normal cornea. More and more researchers consider that pterygium is a tumor rather than degenerative disease. The migration and invasion of tumor cells correlate with the interaction between MMPs and TIMPs. Matrix metalloproteinases (MMPs) are a family of neutral proteolytic enzymes capable of denaturing most components of the extracellular matrix. They are grouped according to their substrate specificity. According to the composed collagen type of cornea, MMP-9 and MMP-10 were assumed to involve the invasion and migration mechanism in pterygium. At the level of inhibition, where MMPs are regulated by their specific tissue inhibitors (TIMPs). Therefore, MMP-9, MMP-10, and TIMP-1 proteins were studied using immunohistochemistry on 82 pterygial specimens and 30 normal conjunctivas. Among the 82 pterygial samples, 29 specimens (35.4%) were positive for MMP-9 expression, 28 were positive for MMP-10 (34.1%), and 59 were positive for TIMP-1 (72.0%). Staining for MMPs was limited to the cytoplasm of the epithelial layer. The TIMP-1 staining was detected in the pterygium epithelium, fibroblasts and corneal epithelium. For further investigate if the invasion ability change after inhibiting TIMPs, pterygium epithelial cells (PECs) were cultured in a serum-free culture medium, and siRNA were used to knock down TIMP gene expression to evaluate the differential ability of pterygium invasion. In the cell model, cell invasion and migration ability increased in TIMP-1 knockdown PECs compared with the parental control. In conclusion, MMP-9 and MMP-10 may each play a role in pterygium formation, and TIMP-1 may contribute to pterygium invasion inhibition.