Ke Y, Jing C, Barraclough R, Smith P, Davies M P, Foster C S
Department of Pathology, University of Liverpool, UK.
Int J Cancer. 1997 May 29;71(5):832-7. doi: 10.1002/(sici)1097-0215(19970529)71:5<832::aid-ijc22>3.0.co;2-8.
Northern and Western blotting techniques were used to study expression of the mRNA and corresponding protein product of the S100-related calcium-binding molecule p9Ka in 6 different metastatic cell lines of the Dunning R3327 rat prostate cancer model. In cells with the lowest metastatic capability (G cells), p9Ka mRNA was barely detectable. In 2 weakly metastatic cell lines (AT-1 and AT-2), p9Ka transcript amounts were, respectively, 6.29 +/- 0.74 and 5.55 +/- 1.11 times that detected in the G cells. In 3 highly metastatic cell lines (AT-3, MAT-LyLu and MAT-Lu), the amounts of p9Ka mRNA were, respectively, 12.85 +/- 2.82, 13.06 +/- 1.69 and 11.62 +/- 1.81 times that expressed in the G cells. Western blot analyses detected no p9Ka protein in the G cells. The amounts of p9Ka protein expressed by tumour cells of intermediate metastatic capability (AT-1 and AT-2) were 3.4 +/- 1.3 microg and 3.3 +/- 1.4 microg, respectively, per 1 x 10(6) cells. The amounts of p9Ka protein expressed by the tumour cells of highest metastatic capability (AT-3, MAT-LyLu and MAT-Lu) were 8.3 +/- 1.1 microg, 8.7 +/- 1.6 microg and 9.6 +/- 1.7 microg, respectively, per 1 x 10(6) cells. Our data reveal a direct association between the elevated expression of mRNA and the p9Ka protein amounts and the increased metastatic capability of individual prostatic cancer cell lines. We suggest that calcium-binding protein p9Ka may play an important role in the metastatic behaviour of rat prostate cancer.
运用Northern印迹法和Western印迹法研究了S100相关钙结合分子p9Ka的mRNA及其相应蛋白质产物在Dunning R3327大鼠前列腺癌模型的6种不同转移细胞系中的表达情况。在转移能力最低的细胞（G细胞）中，几乎检测不到p9Ka mRNA。在2种低转移细胞系（AT-1和AT-2）中，p9Ka转录本量分别是G细胞中检测到量的6.29±0.74倍和5.55±1.11倍。在3种高转移细胞系（AT-3、MAT-LyLu和MAT-Lu）中，p9Ka mRNA量分别是G细胞中表达量的12.85±2.82倍、13.06±1.69倍和11.62±1.81倍。Western印迹分析在G细胞中未检测到p9Ka蛋白。转移能力中等的肿瘤细胞（AT-1和AT-2）每1×10⁶个细胞表达的p9Ka蛋白量分别为3.4±1.3μg和3.3±1.4μg。转移能力最强的肿瘤细胞（AT-3、MAT-LyLu和MAT-Lu）每1×10⁶个细胞表达的p9Ka蛋白量分别为8.3±1.1μg、8.7±1.6μg和9.6±1.7μg。我们的数据揭示了mRNA表达升高及p9Ka蛋白量与各个前列腺癌细胞系转移能力增强之间存在直接关联。我们认为钙结合蛋白p9Ka可能在大鼠前列腺癌的转移行为中起重要作用。
