【摘要】： 目的：研究腫瘤壞死因子-α(TNF-α)、基質(zhì)金屬蛋白酶-2(MMP-2)及細(xì)胞周期蛋白依賴性激酶抑制因子16(P16)在中耳膽脂瘤上皮中的表達(dá)情況,探討它們?nèi)咴谥卸懼錾掀ぶ械谋磉_(dá)有無(wú)相關(guān)性及表達(dá)水平對(duì)中耳膽脂瘤上皮侵襲能力的影響。 方法：標(biāo)本來(lái)自山西醫(yī)科大學(xué)第一臨床醫(yī)學(xué)院耳鼻咽喉-頭頸外科,收集膽脂瘤型中耳炎病人手術(shù)新鮮標(biāo)本,迅速固定,制成石蠟切片,同時(shí)觀察病人的骨質(zhì)破壞程度。采用免疫組化技術(shù)和計(jì)算機(jī)圖像定量分析系統(tǒng)法,檢測(cè)探討腫瘤壞死因子-α(tumor necrosis factor-alpha,TNF-α)、基質(zhì)金屬蛋白酶-2(Matrix metalloproteinases-2,MMP-2)與P16在30例膽脂瘤中的表達(dá),同時(shí)觀察三者的表達(dá)與骨質(zhì)破壞程度的相互關(guān)系,并以10例外耳道皮膚(external ear canal skin,EACS)作為對(duì)照,探討膽脂瘤骨質(zhì)破壞的可能發(fā)生機(jī)制。 結(jié)果： 1.TNF-α、MMP-2與P16在30例中耳膽脂瘤上皮中表達(dá)的陽(yáng)性率分別為83%、76%、63%,與正常外耳道皮膚相比表達(dá)均有顯著性差異(P0.05)。TNF-α、MMP-2表達(dá)陽(yáng)性結(jié)果為棕黃色顆粒,主要定位于胞漿,TNF-α表達(dá)于膽脂瘤上皮細(xì)胞及上皮下結(jié)締層基質(zhì)細(xì)胞,MMP-2在膽脂瘤上皮各層細(xì)胞中均有表達(dá),基底層表達(dá)最強(qiáng),棘層次之,顆粒層及角質(zhì)層細(xì)胞表達(dá)較弱。P16在胞核、胞漿以棕黃色或褐色顆粒為陽(yáng)性表達(dá),并以胞核為主,位于上皮全層,以角化層、顆粒層表達(dá)密度高。 2.膽脂瘤上皮中TNF-α和MMP-2表達(dá)呈正相關(guān)(r=0.694,P0.01)；而P16與TNF-α和MMP-2的表達(dá)無(wú)明顯相關(guān)性(均P0.05)。 3.膽脂瘤侵襲能力與TNF-α和MMP-2表達(dá)有顯著相關(guān)性(均P0.01)。TNF-α和MMP-2表達(dá)灰度值越低,表達(dá)密度越高,膽脂瘤侵襲能力越強(qiáng)。P16在膽脂瘤中的表達(dá)與侵襲能力之間無(wú)相關(guān)性(P0.05)。 結(jié)論： 1.TNF-α、MMP-2與P16在成人中耳膽脂瘤中呈高表達(dá),TNF-α、MMP-2的表達(dá)有相關(guān)性,提示二者之間存在相互作用,后者可能是前者的誘導(dǎo)因素。 2.TNF-α和MMP-2的表達(dá)水平與膽脂瘤的侵襲能力有顯著相關(guān)性,提示TNF-α和MMP-2可能與膽脂瘤的骨質(zhì)破壞程度密切相關(guān)。P16在中耳膽脂瘤中侵襲能力與P16的表達(dá)灰度之間未發(fā)現(xiàn)相關(guān)性。 3.膽脂瘤中TNF-α、MMP-2的高表達(dá)與骨質(zhì)破壞程度有關(guān)指標(biāo)的檢測(cè)可為臨床保守治療、評(píng)估預(yù)后提供參考。 4.P16在膽脂瘤上皮的高度表達(dá)有可能是抑制上皮高度增殖的原因之一。
[Abstract]:Objective: to study the expression of tumor necrosis factor- 偽 (TNF- 偽), matrix metalloproteinase-2 (MMP-2) and cyclin dependent kinase inhibitor 16 (P16) in middle ear cholesteatoma. To investigate the relationship between the expression of these three proteins in middle ear cholesteatoma epithelium and the effect of their expression level on the invasion ability of middle ear cholesteatoma epithelium. Methods: the specimens were collected from the Department of Otorhinolaryngology and head and neck surgery of the first College of Clinical Medicine of Shanxi Medical University. Fresh specimens of cholesteatoma type otitis media were collected, fixed quickly and made into paraffin sections, and the degree of bone destruction was observed at the same time. Tumor necrosis factor- 偽 (tumor necrosis factor-alpha,TNF- 偽 and matrix metalloproteinase-2 (Matrix metalloproteinases-2,) were detected by immunohistochemical technique and computer image quantitative analysis system. The expression of MMP-2 and P16 in 30 cases of cholesteatoma was observed. The relationship between the expression of P16 and the degree of bone destruction was observed, and 10 cases of skin (external ear canal skin,EACS of external auditory canal were used as control. To explore the possible mechanism of bone destruction in cholesteatoma. Results: 1. The positive rates of TNF- 偽, MMP-2 and P16 in the epithelium of 30 cases of middle ear cholesteatoma were 837 ~ 7663, respectively. There were significant differences between TNF- 偽, MMP-2 and P16 in normal skin of external auditory canal (P0.05). The positive expression of MMP-2 was brown granules, mainly located in cytoplasm, TNF- 偽 was expressed in epithelial cells of cholesteatoma and stromal cells in subepithelial connective layer, MMP-2 was expressed in all layers of cholesteatoma epithelium, the expression of MMP-2 in basal layer was the strongest. The expression of P16 was weak in the granular layer and keratinocytes, and the positive expression of P16 in the cytoplasm was brownish yellow or brown granules, and mainly in the nucleus, which was located in the whole layer of the epithelium, with the keratosis layer and the granular layer with high expression density. 2. There was a positive correlation between TNF- 偽 and MMP-2 expression in cholesteatoma epithelium (r 0.694U P0.01), but no significant correlation between P16 expression and TNF- 偽 and MMP-2 expression (P0.05). 3. There was a significant correlation between the invasion ability of cholesteatoma and the expression of TNF- 偽 and MMP-2 (P0.01). The lower the gray value of TNF- 偽 and MMP-2 expression, the higher the density of TNF- 偽 and MMP-2 expression. There was no correlation between the expression of P16 in cholesteatoma and invasion ability (P0.05). Conclusion: 1. TNF- 偽, MMP-2 and P16 are highly expressed in adult middle ear cholesteatoma, and the expression of TNF- 偽 and MMP-2 is correlated, suggesting that there is interaction between them, and the latter may be the inductive factor of the former. 2. 2. The expression levels of TNF- 偽 and MMP-2 were significantly correlated with the invasiveness of cholesteatoma. It is suggested that TNF- 偽 and MMP-2 may be closely related to the degree of bone destruction in cholesteatoma, but there is no correlation between the invasion ability of P16 in middle ear cholesteatoma and the gray level of P16 expression. 3. The high expression of TNF- 偽 and MMP-2 in cholesteatoma may provide reference for clinical conservative treatment and prognosis evaluation. The high expression of 4.P16 in the epithelium of cholesteatoma may be one of the reasons to inhibit the high proliferation of the epithelium.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R764

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