【作者】 曲鑫；

【导师】 曲元明； 徐广明；

【作者基本信息】 山东大学 ， 外科学， 2008， 硕士

【摘要】 目的研究CD147、基质金属蛋白酶-9及血管内皮生成因子在垂体腺瘤中的蛋白质表达,探讨CD147、MMP-9及VEGF与垂体腺瘤侵袭发生的相关性以及它们之间的内在联系。方法1.标本来源收集2006、2～2007、4期间山东省立医院神经外科的垂体腺瘤手术标50例,其中男性26例,女性24例,年龄18～71岁,平均年龄38.3±1.3岁;所有病例诊断均有术后的免疫组化病理证实。依据影像学提示肿瘤侵犯海绵窦或有骨质破坏,和/或术中观察肿瘤对周围骨质、海绵窦有/无侵犯等情况分为侵袭性组(29例)和非侵袭性组(21例),根据分泌功能分类:功能性垂体腺瘤(泌乳素腺瘤15例、生长激素腺瘤6例、促肾上腺腺瘤2例、多功能腺瘤8例)和无功能性垂体腺瘤(19例)。2.免疫组化染色方法采用免疫组化链霉菌抗生物素蛋白-过氧化物酶连接(S-P)法,按标准实验流程,检测CD147、MMP-9、VEGF在侵袭性和非侵袭性垂体瘤中的蛋白质表达。3.结果判定标准CD147以肿瘤细胞质或胞膜出现黄色或黄褐色颗粒为阳性,MMP-9以肿瘤细胞胞质内出现棕黄色颗粒为阳性,VEGF以肿瘤细胞浆出现棕黄色颗粒为阳性。在高倍视野(×400)下,每张切片随机观察5个视野的肿瘤细胞,计算200个肿瘤细胞免疫反应阳性的细胞百分比,根据CD147、MMP-9及VEGF反应阳性细胞显色强度及范围进行评价:①阴性(一):无阳性细胞染色;②弱阳性(+):阳性细胞小于25%或显色浅;③中度阳性(++):阳性细胞在26%～50%或染色略深;④强阳性(+++):阳性细胞大于50%或显色深。结果1、免疫组化显色结果:1)、CD147在侵袭性和非侵袭性垂体腺瘤中强阳性表达率分别为51.7%和23.8%,阳性表达率分别为93.1%和52.4%,两组比较差异均具有统计学意义(x~2=3.95,P＜0.05和x~2=11.07,P＜0.05)。2)、MMP-9在侵袭性和非侵袭性垂体腺瘤中的强阳性表达率分别为58.6%和14.3%,阳性表达率分别为86.2%和42.9%,两组比较差异具有统计学意义(x~2=9.98,P＜0.05和x~2=10.52,P＜0.05)。3)、VEGFF在侵袭性和非侵袭性垂体腺瘤中强阳性表达率为62.1%和19.1%,阳性表达率为96.6%和57.1%,两组比较差异具有统计学意义(x~2=9.15,P＜0.05和x~2=11.82,P＜0.05)。2、相关分析:Pearson相关分析显示,侵袭性垂体腺瘤中CD147与MMP-9表达成正相关(r_s=0.373,P＜0.05),CD147与VEGF表达成正相关(r_s=0.509,P＜0.01),MMP-9与VEGF表达成正相关(r_s=0.781,P＜0.01);而非侵袭性垂体腺瘤组中CD147、MMP-9与VEGF的蛋白质表达无相关性(P＞0.05)。结论1、CD147、MMP-9、VEGF在侵袭性与非侵袭性垂体腺瘤中的表达有显著性差异。侵袭性垂体腺瘤中CD147、MMP-9、VEGF蛋白表达较非侵袭性垂体腺瘤显著增高。2、垂体腺瘤的侵袭性生长可能与CD147、MMP-9、VEGF高表达有关,在肿瘤侵袭生长过程中,CD147与MMP-9的产生有关,VEGF与MMP-9的产生有关,CD147与VEGF的生长有关,共同促进垂体腺瘤侵袭性生长。3、CD147、MMP-9、VEGF可以作为判断垂体腺瘤侵袭性生长的有效指标。4、患者性别及垂体腺瘤有无分泌功能与垂体腺瘤侵袭性发生率无关。更多还原

【Abstract】 ObjectiveThe aim is to study the protein expression of CD147、MMP-9 and VEGF in pituitary adenomas, and to study the biological relationship between CD147、MMP-9、VEGF and the invasiveness of pituitary adenomas, and to evaluate the clinical significance.Method1、Patient SelectionTissue samples were obtained from patients who underwent surgery in Shandong Provincial Hospital from February, 2006 to April, 2007. The total number of tissue samples was 50, including 26 males and 24 females with an average age of 38.3±1.3. All diagnoses of these cases were verified by immunohistochemical pathology. According to the illustration of images and observation in operations, cases were divided into 29 invasive cases (PRL 5 cases, GH 6 cases, ACTH 2 ones , and multifuctional adenomas 8 cases)and 21 noninvasive cases(19 cases); according to secreting function, cases were divided into 31 functional cases and 19 nonfunctional cases.2、ImmunohistochemistryThe protein expression of CD147, MMP-9 and VEGF were detected by the S-P method with ultrasensitive S-P kit.3、Result determinationThe positive staining of CD147 is yellow or yellow brown particles located in cytoplasm or cellular membrane. The positive staining of MMP-9 or VEGF is yellow brown particles located in cytoplasm. 5 fields at a magnification of 400 X at every slice were observed at random .The percentage of positive staining cells in 200 tumor cells was calculated. The results were evaluated by staining intense and extent in positive cells:①negative(-): no staining;②weak positive(+):the number of positive cells < 25% or slight staining;③mild positive(++):the number of positive cells from 26% to 50% or mild staining;④strong positive(+++): the number of positive cells > 50% or deep staining.ResultsThe rates of intense positive expression of CD147 in invasive and noninvasive cases were 54% and 20.7% respectively; the rates of total positive expression were 96.8% and 51.4%; and there were statistic significance in the two teams respectively( P<0.05). The rates of intense positive expressoin of MMP-9 in invasive and noninvasive ones were 58.6% and 14. 3% respectively; the total positive expression were 86. 2% and 42. 9% respectively; and there were statistic significance in the two teams respectively( P<0.05). The rates of intense positive expressoin of VEGF in invasive and noninvasive ones were 62.1% and 19.1% respectively; the total positive expression were 96.6% and 57.1% respectively; and there were statistic significance in the two teams respectively( P<0.05). There was a significant positive correlation among protein expression of CD147, MMP-9 and VEGF in invasive cases; but there were not significant correlation in noninvasive cases. And the incidence of the invasiveness has no significant difference with the sexulty of patients and secretion of adenomas .ConclusionThe protein expression of CD147, MMP-9 and VEGF may play important roles in the invasiveness of pituitary adenomas and may be useful markers to predict the invasiveness. The invasiveness growth of adenomas may has connection with high protein expression of CD147, MMP-9 and VEGF. During invasive growth , there are some correlation among CD147, MMP-9 and VEGF; reciprocal promotion between CD147 and MMP-9,MMP-9 and VEGF. And the incidence of the invasiveness has no significant difference with the sexulty of patients and secretion of adenomas .更多还原