Pajouh M S, Nagle R B, Breathnach R, Finch J S, Brawer M K, Bowden G T
Department of Microbiology and Immunology, University of Arizona Medical School, Tucson 85724.
J Cancer Res Clin Oncol. 1991;117(2):144-50. doi: 10.1007/BF01613138.
Twenty-five surgical specimens of malignant human prostate, 3 lymph nodes with metastatic prostate carcinoma, 11 normal human prostates, as well as 3 human prostate cell lines (DU-145, PC3 and LNCaP) were examined for the expression of the human matrix metalloproteinase-7 gene (MMP-7) from the human collagenase family (originally called PUMP-1 for putative metalloproteinase-1) [Quantin et al. (1989) Biochemistry 28:5327-5334; Muller et al. (1988) Biochem J 253:187-192; Matrisian and Bowden (1990) Semin Cancer Biol 1:107-115]. Northern blots were prepared using total RNA extracted from 18 prostate adenocarcinomas, 2 lymph nodes with metastatic prostate carcinoma and 11 normal human prostates. When the northern blots were hybridized with a 32P-labeled MMP-7 cDNA probe, a 1.2-kb mRNA was detected in 14 out of 18 prostate adenocarcinomas, 1 out of 2 metastatic lymph nodes, and 3 out of 11 normal prostates. The 3 human prostate cell lines did not show any evidence of the MMP-7 transcript. In situ hybridization was conducted to localize the MMP-7 mRNA to individual cells using a 35S-labeled MMP-7 cRNA. In situ hybridization was carried out on snap-frozen tissue sections of 7 prostate adenocarcinomas and 3 lymph nodes containing metastatic prostate adenocarcinoma using the same tissues previously probed by northern analysis as well as new samples. In situ hybridization revealed that the MMP-7 gene was expressed in the epithelial cells of primary prostate adenocarcinoma as well as in invasive and metastatic cells. MMP-7 expression was also seen focally in some dysplastic glands but not in stroma. Additional northern blot analysis was performed using probes to human type-IV collagenase, type-I collagenase and stromelysin I in human prostate adenocarcinoma as well as normal prostate tissue. Our results indicated that 6 out of 10 adenocarcinoma samples and none of the 4 normal samples were positive for type-IV collagenase transcripts. Tissue samples were also examined for the expression of type-I collagenase (9 adenocarcinomas and 4 normal) and stromelysin I (13 adenocarcinomas) by northern analysis. None of the tissues was found to express the transcripts of interest at detectable levels. These data suggest that certain metalloproteinases are present in prostatic adenocarcinoma and may play a role in invasion and metastasis.
对25例人类恶性前列腺手术标本、3例伴有转移性前列腺癌的淋巴结、11例正常人类前列腺以及3种人类前列腺细胞系(DU - 145、PC3和LNCaP)进行了检测,以分析人类胶原酶家族中人类基质金属蛋白酶 - 7基因(MMP - 7,最初称为PUMP - 1,即假定的金属蛋白酶 - 1)的表达情况[Quantin等人(1989年)《生物化学》28:5327 - 5334;Muller等人(1988年)《生物化学杂志》253:187 - 192;Matrisian和Bowden(1990年)《癌症生物学研讨会》1:107 - 115]。使用从18例前列腺腺癌、2例伴有转移性前列腺癌的淋巴结和11例正常人类前列腺中提取的总RNA制备Northern印迹。当Northern印迹与32P标记的MMP - 7 cDNA探针杂交时,在18例前列腺腺癌中的14例、2例转移性淋巴结中的1例以及11例正常前列腺中的3例中检测到了1.2 kb的mRNA。3种人类前列腺细胞系未显示出MMP - 7转录本的任何证据。使用35S标记的MMP - 7 cRNA进行原位杂交,将MMP - 7 mRNA定位到单个细胞。使用之前经Northern分析检测过的相同组织以及新样本,对7例前列腺腺癌和3例含有转移性前列腺腺癌的淋巴结的速冻组织切片进行原位杂交。原位杂交显示,MMP - 7基因在原发性前列腺腺癌的上皮细胞以及侵袭性和转移性细胞中表达。在一些发育异常的腺体中也局部观察到MMP - 7表达,但在基质中未观察到。使用针对人类IV型胶原酶、I型胶原酶和基质溶解素I的探针,对人类前列腺腺癌以及正常前列腺组织进行了额外的Northern印迹分析。我们的结果表明,10例腺癌样本中有6例IV型胶原酶转录本呈阳性,而4例正常样本均为阴性。还通过Northern分析检测了组织样本中I型胶原酶(9例腺癌和4例正常样本)和基质溶解素I(13例腺癌)的表达。未发现任何组织在可检测水平表达感兴趣的转录本。这些数据表明,某些金属蛋白酶存在于前列腺腺癌中,可能在侵袭和转移中起作用。