基质金属蛋白酶抑制剂4在皮肤恶性黑素瘤中的表达及其与肿瘤发生、侵袭和转移的相关性

【摘要】 目的 探讨基质金属蛋白酶抑制剂4（TIMP-4）在皮肤恶性黑素瘤（CMM）中的表达及其与肿瘤增殖、侵袭和转移的相关性。 方法 应用蛋白免疫印迹方法分析5例CMM和瘤旁组织及3例色素痣组织中TIMP-4蛋白表达差异。应用免疫组化法检测TIMP-4在43例CMM和51 例色素痣组织中的表达，并分析其与临床病理特征、Ki-67、基质金属蛋白酶2（MMP-2）、血管内皮细胞生长因子（VEGF）及黑素瘤表面标志物CD63表达的相关性。免疫组化法检测Ki-67、MMP-2、VEGF和CD63表达。 结果 Western印迹结果提示，5例CMM组织中4例TIMP-4表达高于瘤旁组织和3例色素痣组织。免疫组化分析结果，TIMP-4在43例CMM和51 例色素痣中分别有37例和10例阳性，阳性率分别为86.04%和19.6%，两组差异有统计学意义（χ2 = 31.55，P < 0.05）。TIMP-4表达水平与CMM的进展呈正相关（rs = 0.309，P < 0.05），随着肿瘤从原位到侵袭性再发展到转移性，TIMP-4表达水平呈升高趋势。CMM中TIMP-4表达与临床分期、肿瘤厚度、Clark分级、溃疡与否等预后变量无显著相关性，与Ki-67亦无显著相关，但TIMP-4表达与VEGF表达呈正相关（rs = 0.345，P < 0.05）；CMM中MMP-2阳性组TIMP-4的表达明显高于MMP-2阴性组（P < 0.01）；此外，TIMP-4表达与CD63表达呈正相关（rs = 0.555，P < 0.01）。 结论 TIMP-4在CMM中呈高表达，可能参与CMM的发生及发展，尤其与其转移及血管生成密切相关。 【关键词】 黑色素瘤； 金属蛋白酶类组织抑制剂； 基质金属蛋白酶类

Expression of tissue inhibitor of metalloproteinase-4 in human cutaneous malignant melanoma tissue and its relationship with melanoma initiation,invasion and metastasis

HUANG Ying-xue, ZHOU Lu, ZHANG Wei, SHAO Xue-bao, LI A-mei, XU Xiu-lian，SUN Jian-fang. Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing 210042, ChinaCorresponding authors: XU Xiu-lian, Email: xxlqjl@sina.com; SUN Jian-fang, Email: fangmin5758@yahoo.com.cn 【Abstract】 Objective To detect the expression of tissue inhibitor of metalloproteinase-4（TIMP-4） in cutaneous malignant melanoma （CMM） tissue and to assess its relationship with melanoma proliferation, invasion and metastasis. Methods Western blot was conducted to measure the protein expression of TIMP-4 in five fresh lesional and paratumoral tissue specimens of CMM and three fresh tissue specimens of nevi. Immunohistochemistry was carried out to quantify the expression of TIMP-4, Ki-67, matrix metalloproteinase-2 （MMP-2）, vascular endothelial growth factor （VEGF） and CD63 in paraffin-embedded tissue samples from 43 cases of CMM and 51 cases of nevi. The degree of malignancy of melanoma was evaluated in these lesions. Results Western blot analysis showed that the expression of TIMP-4 was significantly higher in 4 of 5 CMM tissue specimens than in corresponding paratumoral tissue specimens and nevus tissue specimens. Immunohistochemistry revealed that the expression rate of TIMP-4 was 86.04% （37/43） in melanoma tissue, compared to 19.6% （10/51） in nevus tissue （χ2 = 31.55, P < 0.05）. The expression of TIMP-4 increased sequentially from in situ melanoma to invasive and metastatic melanoma （rs = 0.309, P < 0.05）. As far as CMM was concerned, the TIMP-4 expression was uncorrelated with any of the known prognostic variables including clinical stage, Clark level, Breslow depth, presence of ulcer, and Ki-67 expression （all P > 0.05）, but positively correlated with the expressions of VEGF （rs = 0.345, P < 0.05） and CD63 （rs = 0.555，P < 0.01）. The median expression level of TIMP-4 was significantly higher in MMP-2-positive than in MMP-2-negative melanoma tissue samples （3 vs. 0, P < 0.01）. Conclusions TIMP-4 protein is highly expressed in CMM tissue, which may be closely associated with the initiation and progression of CMM, especially with the metastasis of and angiogenesis in CMM. 【Key words】 Melanoma; Tissue inhibitor of metalloproteinases; Matrix metalloproteinases