【作者】 刘胜平；

【导师】 宗世璋；

【作者基本信息】 江西医学院 ， 外科学， 2005， 硕士

【摘要】 骨肉瘤(Osteosarcoma)是最常见的原发恶性骨肿瘤,占原发恶性骨肿瘤的20%,多好发于青少年,其生物学行为多呈高度恶性,早期即易发生血行性转移。1970 年以前标准治疗是截肢术,但五年生存率在10%。近20余年来,随着新辅助化疗广泛应用和手术方法的改进,骨肉瘤五年生存率已达到80%以上,但仍有相当一部分病人晚期死于肺部转移。有文献报道:80%的骨肉瘤病人在确诊时即已发生了肺部转移,是骨肉瘤病患者晚期死亡的主要原因。因此,开展对骨肉瘤的微转移机制的研究对于提高骨肉瘤的治疗效果及判定预后具有一定意义。血管内皮生长因子(VEGF)在肿瘤生长和转移的过程中发挥重要的作用,而肿瘤转移抑制基因(nm23)却有抑制肿瘤转移的作用。其两者在骨肉瘤的联合表达作用目前研究报道较少。目的:探讨VEGF、nm23在骨肉瘤中的表达及其生物学意义,并对其相互之间关系进行研究,为阐明骨肉瘤的微转移机制和了解预后提供一定的理论依据。方法:本试验以江西医学院第一附属医院1998 年1 月～2003 年12 月手术切除或活检的40 例骨肉瘤患者的病理标本及临床资料为研究对象。应用SABC免疫组化检测CD34、VEGF 和nm23 单克隆抗体在40 例骨肉瘤标本的表达情况。阴性对照用PBS 代替一抗CD34、VEGF 和nm23 抗体,阳性对照以福州迈新公司购买的标准阳性片作为对照。VEGF、nm23 以细胞浆内出现棕黄色至棕深色颗粒作为阳性判断标准,即阴性(-)为更多还原

【Abstract】 Osteosarcoma (OS) is the most common primary malignant bone tumor inadolescents and young adults, comprising 20% of all such malignancies. Itsbiological behavior is always highly malignant. Its hematogenous metastasisoccurs more frequently and early. The standard therapy of OS is amputationbefore 1970, but the 5-year survival rate of OS is less 10%. Despite recentadvances in multimodality treatments consisting of aggressively adjuvantchemotherapy and wide tumor excision make the 5-year survival rate of OSclimb to 80%, a significant percentage of patients die from lung metastasis.Some papers report that 80 percent of osteosarcoma patients have lungmetastasis in final diagnosis, which is the first cause of death of advancedosteosarcoma. So it is a great significance to research the mechanism ofmicro-metastasis of OS in improving the clinical results of therapy andjudging the prognosis. VEGF makes great function in stimulating the tumorgrowth and making it metastasis, but nm23 can anti-metastasis. Theco-research of VEGF and nm23 in OS is seldom reported.Objective: To investigate the expression and function of vascular endothelialgrowth factor (VEGF) and nm23 and research their relations in osteosarcoma,which provides theoretical foundation for explaining the mechanism ofmicro-metastasis and comprehending the prognosis of OS.Method: From Jan. 1998 to Dec. 2003, 40 osteosarcoma patients treated inthe first affiliated hospital of Jiangxi medical college, with follow-up morethan two years, were selected to enter this study. Specimens of 40 OS werestudied by immunohistochemical S-ABC method to VEGF, CD34 and nm23.The known standard positive specimen from Fuzhou Maixin biological Co.Ltd. was the positive contract, and the negative contract was PPS instead ofVEGF, CD34 and nm23 as the first antibody in this experiment. Expression ofVEGF, nm23 gene is that the cell dyed brown and obvious dense backgrounddying is named positive. And the number of positive cells was counted fromwhich one hundred of cells in high-scope with incompletely randomizedprinciple. When the number of the positive cells are less than 24,theexpression rate is recorded(+),then the number of positive cells are between25-49, 50-74, 75-100, and the expression rates are(++),(+++),(++++)respectively. The high expression of VEGF and nm23 is (+++)to(++++),andthe low expression is(+)to(++). Statistic soft file of SPSS 10.0 for Windowswere used in Statistics analysis. All the enumeration data was analyzed byFisher’s exact test and the measurement data was analyzed by student t test.Results: VEGF staining was positive in 37 tumors (92.5%) which theCD34-positive micro-vessels was significantly higher than that inVEGF-negative 3 tumors (P<0.05), and multivariate analysis indicated thatmicro-vessel density (MVD) and VEGF expression were closely correlatedwith patients’poor prognosis as well as metastasis. There was no correlationbetween patient’s prognosis and patient’s gender, age, the time of illness.nm23 staining was positive in 18 tumors (45%), which was no significancebetween patient’s prognosis and the expression of nm23 in statistics.Conclusion: VEGF is a critical angiogenic stimuli which stimulatesangiogenesis, tumor growth and metastasis in osteosarcoma, the microvesseldensity and VEGF expression can function as two independent prognosticvariables in osteosarcoma. But nm23 is nonmetastasis suppressor inosteosarcoma, which expression can’t play important role in prognosis ofosteosarcoma.更多还原