作者：陈述伟,杨述华,张劲松,朱续胜，袁永辉
【摘要】 目的 探讨骨巨细胞瘤中的PTEN、MMP7、VEGF表达与“三结合分类”、病理分级及复发、转移的关系。方法 采用免疫组化法检测65例骨巨细胞瘤标本PTEN、MMP7、VEGF基因蛋白的表达，分析其与骨巨细胞瘤临床病理分期、分级及复发、转移的关系。结果 PTEN、MMP7、VEGF的阳性表达率分别为56.95%、72.30%、49.20%；在“三结合分类”组中PTEN的阳性表达率分别为83.3%、68.10%、28.00%，呈下降趋势，良性与中间性组间差异不显著(P=0.271)，而中间性与恶性组间差异明显(P=0.001)；而MMP7阳性表达率分别为39.00%、72.50%、96.00%，VEGF阳性表达率分别为16.67%、54.50%、68.00%，组间差异明显(P=0.031、0.001)；在转移组中PTEN、MMP7、VEGF的阳性表达率分别为30.00%、100%、85.00%，在无转移组中PTEN、MMP7、VEGF的阳性表达率分别为68.80%、60.00%、33.3%，两组间差异明显（P=0.003、0.001、0.001）；复发组中PTEN、MMP7、VEGF的阳性表达率分别为33.30%、93.30%、86.60%，无复发组中PTEN、MMP7、VEGF的阳性表达率分别为64.00%、66.00%、38.00%，两组间差异明显（P=0.035、0.038、0.001），且PTEN与MMP7、VEGF负相关（P=0.001、0.020），而与病理分级及年龄、性别无统计学差异。结论 PTEN、MMP7、VEGF的表达与“三结合分类”有一定的关系，并与转移、复发显著相关，联合检测有助于骨巨细胞瘤的临床评估，对其预后判断具有重要临床意义。
【关键词】 骨巨细胞瘤；PTEN；MMP7；VEGF；免疫组织化学
Abstract：Objective To evaluate the expressions of PTEN ( phosphatase and tension homologue deleted on chromosome 10) ，MMP7(matrix metalloproteinases7，MMP7)，VEGF （vascular endothelial growth factor） and their relationships with clinical factors，pathological types and metastasis，recurrence of giant cell tumor of bone (GCT).Methods The expressions of PTEN， MMP7， VEGF in 65 cases of human GCT were detected by using immunohistochemical staining. The expressions of PTEN， MMP7， VEGF and other clinical fractures were analyzed with the metastasis and recurrence of the tumor. Results The expressions of the PTEN，MMP7，VEGF in GCT were 56.95% and 72.30%，49.20%, respectively. In the clinicalpathologicalXray groups, the expressions of PTEN were 83.30%，68.10%， 28.00%,the positive rate had no significantly difference between the Nonmalignant and doutful malignant groups, P＞0.05(P=0.271), but there were significantly difference between the doutful malignant and malignant groups, P＜0.05(P=0.001) ；the expression of MMP7 were 39.00%，72.50%，96.00%,and the expression of VEGF were 16.67%，54.50%，68.00%,the positive rates of them had significantly difference, P＜0.05 (P=0.031,0.001)；the expressions of PTEN ，MMP7，VEGF in the recurrence group were 33.30%，93.30%，86.60%, but in the nonrecurrence group were 64.00%，66.00%，38.00%,their expressions were statistically related to the recurrence of GCT, P＜0.05 (P=0.035, 0.038,0.001); in the metastasis group, the expressions of PTEN ，MMP7，VEGF were 30.00%，100%，85.00%,in the no metastasis group ，the positive rates of PTEN，MMP7，VEGF were 68.80%，60.00%，33.30% ,the positive rates of them had significantly difference,P＜0.05（P=0.003，0.001，0.001）;at the same time, The expression of PTEN in GCT was inversely correlated with the expressions of VEGF and MMP7, P＜0.05(P=0.001, 0.02) ; However, the expressions of PTEN 、 MMP7、VEGF had no statistically significance with the pathological types and sexuality.Conclusion The decreased expression of PTEN and increased expressions of MMP7、VEGF have significant relationship to the prognosis of GCT. The combined detection of PTEN and MMP7 may be of important clinical value to evaluate the infiltrative ability and prognosis of GCT.
Key words：Giant cell tumor of bone(GCT)； PTEN/biosynthesis； MMP7/biosynthesis；VEGF/biosynthesis; Immunohistochemistry
0 引言
骨巨细胞瘤(Giant cell tumors of bone，GCT)是一种中间的，潜在的恶性肿瘤，其生物学行为复杂而多变，易发生转移与复发。与肿瘤细胞的增殖调控、基底膜的降解、血管的增生密切相关。我们应用免疫组织化学方法，联合检测PTEN、MMP7、VEGF在骨巨细胞瘤中的表达，探讨其与肿瘤细胞的生长、增殖、浸润和转移等生物学行为及其与临床病理分期分级的关系，旨在为治疗GCT与评估预后提供理论依据。
1 资料与方法
1.1 样本来源
选择1995年8月25日～2004年8月23日华中科技大学同济医学院附属协和医院（48例）、同济医学院病理科（17例），有完整随访资料的手术切除骨巨细胞瘤标本共65例，其中男38例，女27例，年龄15～58岁，平均年龄35.2岁，中位数年龄28岁；单纯病灶刮除12例，切除加阿霉素骨水泥填塞16例，广泛切除及人工关节置换37例；随访时间15～120个月，平均48.3个月；有15例复发，其中单纯病灶刮除组6例，切除加骨水泥组4例，广泛切除组5例；转移20例，其中Ⅰ级3例，Ⅱ级6例，Ⅲ级11例。根据Jafee 1940年提出的病理分级标准 [1]：Ⅰ级18例，Ⅱ级17例，Ⅲ级30例。结合临床、病理、影像学表现，即“三结合分类”，参考WHO（2002）骨肿瘤分类标准 [2]，良性18例，恶性25例，中间性22例。
1.2 主要试剂
PTEN兔抗人多克隆抗体，MMP7、VEGF鼠抗人单克隆抗体，羊抗兔、羊抗鼠SP试剂盒及DAB显示剂，均购自北京中山生物技术公司。
1.3 方法
各种标本常规石蜡包埋，5μm连续切片，分别做HE和免疫组化染色，免疫组化染色为SP法，染色方法依说明书进行，阳性对照采用已知实验中证实对本实验所用抗体呈阳性反应的阳性切片，以PBS代替一抗作阴性对照。
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