The present study aimed to detect comparative expression of integrin αVβ3 and its involvement in expression and activation of matrix metalloproteinase-2 (MMP-2) in 25 malignant human breast tumor and adjacent normal...The present study aimed to detect comparative expression of integrin αVβ3 and its involvement in expression and activation of matrix metalloproteinase-2 (MMP-2) in 25 malignant human breast tumor and adjacent normal breast tissues from different clinical TNM stages (DCIS to T4) of the disease and possible involvement of known regulating parameters of MMP-2 like TIMP-2, MT1-MMP and EMPRIN. Integrin αVβ3 was highly expressed in tumors than adjacent normal breast tissues. Pro-MMP-2(72-KD) was mainly expressed in adjacent normal tissues compared to tumors. The mature forms of MMP-2 (68 KD and 64 KD) were found only in tumors. Appreciable expression of TIMP-2 and induction of MT1-MMP and EMPRIN in T2-T4 stages suggested their possible role in MMP-2 activation. Over expression Integrin αVβ3 in tumors than adjacent normal breast tissues was an indication of cancer progression with involvement of integrin signaling. We conclude that, the co-precipitation of MMP-2 with αvβ3 by anti-αv antibody is a strong indication that integrin αvβ3 is a surface receptor for MMP-2 and αvβ3-MMP-2 complex on the surface of tumor cells may play a very important role in determining the invasive property and malignant behavior of tumor tissues. The positive expression of endogenous inhibitor of MMP-2, TIMP-2 may have an appreciable role in activation of this protease and risk of malignancy in advanced stage of the disease. The enhanced expression of MT1-MMP and EMPRIN suggested a role for these factors in gelatinase regulation. However the exact mechanism(s) remains to be investigated. Finally, evaluation of integrin αVβ3 associated MMP-2 expression and activity may add valuable information and can possibly be therapeutic target. The clinical exploitation of integrins will provide oncologists with novel therapeutic strategies for the treatment of malignancy in breast cancer.展开更多
目的:应用新的噬菌体删减筛选技术筛选能与整合素αvβ_3具特异性结合功能的小肽分子,并简单分析其生物学功能。方法:以高表达整合素αvβ_3分子的M2黑色素瘤细胞株为靶细胞,以低/不表达整合素αvβ_3分子的C23黑色素瘤细胞株为吸附细胞...目的:应用新的噬菌体删减筛选技术筛选能与整合素αvβ_3具特异性结合功能的小肽分子,并简单分析其生物学功能。方法:以高表达整合素αvβ_3分子的M2黑色素瘤细胞株为靶细胞,以低/不表达整合素αvβ_3分子的C23黑色素瘤细胞株为吸附细胞,对噬菌体随机七肽库进行消减筛选,ELISA鉴定阳性克隆,DNA测序和分析。结果:经过4轮亲和筛选,3轮删减筛选,经固定ELISA鉴定,随机选取50个克隆得到2个能特异性与M2细胞结合而不与C23细胞结合的阳性克隆,进行DNA测序后,其多肽序列如下:HTPPYIL;MNFNAML。结论:通过噬菌体噬菌体删减筛选技术获得与整合素alpha v beta 3具特异性结合能力的小肽分子,为基于整合素alpha v beta 3的肿瘤血管生成的早期诊断及治疗的多肽药物前体设计提供实验依据。展开更多
文摘The present study aimed to detect comparative expression of integrin αVβ3 and its involvement in expression and activation of matrix metalloproteinase-2 (MMP-2) in 25 malignant human breast tumor and adjacent normal breast tissues from different clinical TNM stages (DCIS to T4) of the disease and possible involvement of known regulating parameters of MMP-2 like TIMP-2, MT1-MMP and EMPRIN. Integrin αVβ3 was highly expressed in tumors than adjacent normal breast tissues. Pro-MMP-2(72-KD) was mainly expressed in adjacent normal tissues compared to tumors. The mature forms of MMP-2 (68 KD and 64 KD) were found only in tumors. Appreciable expression of TIMP-2 and induction of MT1-MMP and EMPRIN in T2-T4 stages suggested their possible role in MMP-2 activation. Over expression Integrin αVβ3 in tumors than adjacent normal breast tissues was an indication of cancer progression with involvement of integrin signaling. We conclude that, the co-precipitation of MMP-2 with αvβ3 by anti-αv antibody is a strong indication that integrin αvβ3 is a surface receptor for MMP-2 and αvβ3-MMP-2 complex on the surface of tumor cells may play a very important role in determining the invasive property and malignant behavior of tumor tissues. The positive expression of endogenous inhibitor of MMP-2, TIMP-2 may have an appreciable role in activation of this protease and risk of malignancy in advanced stage of the disease. The enhanced expression of MT1-MMP and EMPRIN suggested a role for these factors in gelatinase regulation. However the exact mechanism(s) remains to be investigated. Finally, evaluation of integrin αVβ3 associated MMP-2 expression and activity may add valuable information and can possibly be therapeutic target. The clinical exploitation of integrins will provide oncologists with novel therapeutic strategies for the treatment of malignancy in breast cancer.
文摘目的:应用新的噬菌体删减筛选技术筛选能与整合素αvβ_3具特异性结合功能的小肽分子,并简单分析其生物学功能。方法:以高表达整合素αvβ_3分子的M2黑色素瘤细胞株为靶细胞,以低/不表达整合素αvβ_3分子的C23黑色素瘤细胞株为吸附细胞,对噬菌体随机七肽库进行消减筛选,ELISA鉴定阳性克隆,DNA测序和分析。结果:经过4轮亲和筛选,3轮删减筛选,经固定ELISA鉴定,随机选取50个克隆得到2个能特异性与M2细胞结合而不与C23细胞结合的阳性克隆,进行DNA测序后,其多肽序列如下:HTPPYIL;MNFNAML。结论:通过噬菌体噬菌体删减筛选技术获得与整合素alpha v beta 3具特异性结合能力的小肽分子,为基于整合素alpha v beta 3的肿瘤血管生成的早期诊断及治疗的多肽药物前体设计提供实验依据。
