Perlino E, Lovecchio M, Vacca R A, Fornaro M, Moro L, Ditonno P, Battaglia M, Selvaggi F P, Mastropasqua M G, Bufo P, Languino L R
Center of Study on Mitochondria and Energy Metabolism, Consiglio Nazionele delle Ricerche, Bari, Italy.
Am J Pathol. 2000 Nov;157(5):1727-34. doi: 10.1016/s0002-9440(10)64809-2.
Alterations of integrin expression levels in cancer cells correlate with changes in invasiveness, tumor progression, and metastatic potential. The beta1C integrin, an alternatively spliced form of the human beta1 integrin, has been shown to inhibit prostate cell proliferation. Furthermore, beta1C protein levels were found to be abundant in normal prostate glandular epithelium and down-regulated in prostatic adenocarcinoma. To gain further insights into the molecular mechanisms underlying abnormal cancer cell proliferation, we have studied beta1C and beta1 integrin expression at both mRNA and protein levels by Northern and immunoblotting analysis using freshly isolated neoplastic and normal human prostate tissue specimens. Steady-state mRNA levels were evaluated in 38 specimens: 33 prostatic adenocarcinomas exhibiting different Gleason's grade and five normal tissue specimens that did not show any histological manifestation of benign prostatic hypertrophy. Our results demonstrate that beta1C mRNA is expressed in normal prostate and is significantly down-regulated in neoplastic prostate specimens. In addition, using a probe that hybridizes with all beta1 variants, mRNA levels of beta1 are found reduced in neoplastic versus normal prostate tissues. We demonstrate that beta1C mRNA down-regulation does not correlate with either tumor grade or differentiation according to Gleason's grade and TNM system evaluation, and that beta1C mRNA levels are not affected by hormonal therapy. In parallel, beta1C protein levels were analyzed. As expected, beta1C is found to be expressed in normal prostate and dramatically reduced in neoplastic prostate tissues; in contrast, using an antibody to beta1 that recognizes all beta1 variants, the levels of beta1 are comparable in normal and neoplastic prostate, thus indicating a selective down-regulation of the beta1C protein in prostate carcinoma. These results demonstrate for the first time that beta1C and beta1 mRNA expression is down-regulated in prostate carcinoma, whereas only beta1C protein levels are reduced. Our data highlight a selective pressure to reduce the expression levels of beta1C, a very efficient inhibitor of cell proliferation, in prostate malignant transformation.
癌细胞中整合素表达水平的改变与侵袭性、肿瘤进展和转移潜能的变化相关。β1C整合素是人类β1整合素的一种可变剪接形式,已被证明可抑制前列腺细胞增殖。此外,发现β1C蛋白水平在正常前列腺腺上皮中丰富,而在前列腺腺癌中下调。为了进一步深入了解癌细胞异常增殖的分子机制,我们使用新鲜分离的肿瘤性和正常人类前列腺组织标本,通过Northern印迹和免疫印迹分析,在mRNA和蛋白质水平上研究了β1C和β1整合素的表达。在38个标本中评估了稳态mRNA水平:33个表现出不同Gleason分级的前列腺腺癌和5个未显示任何良性前列腺增生组织学表现的正常组织标本。我们的结果表明,β1C mRNA在正常前列腺中表达,而在肿瘤性前列腺标本中显著下调。此外,使用与所有β1变体杂交的探针,发现肿瘤性前列腺组织与正常前列腺组织相比,β1的mRNA水平降低。我们证明,根据Gleason分级和TNM系统评估,β1C mRNA下调与肿瘤分级或分化均无关,并且β1C mRNA水平不受激素治疗影响。同时,分析了β1C蛋白水平。正如预期的那样,发现β1C在正常前列腺中表达,而在肿瘤性前列腺组织中显著降低;相反,使用识别所有β1变体的β1抗体,β1在正常和肿瘤性前列腺中的水平相当,因此表明前列腺癌中β1C蛋白存在选择性下调。这些结果首次证明,前列腺癌中β1C和β1 mRNA表达下调,而只有β1C蛋白水平降低。我们的数据突出了在前列腺恶性转化过程中,降低细胞增殖非常有效的抑制剂β1C表达水平的选择性压力。