De Luca A, Casamassimi A, Selvam M P, Losito S, Ciardiello F, Agrawal S, Salomon D S, Normanno N
Oncologia Sperimentale D, ITN-Fondazione Pascale, Naples, Italy.
Int J Cancer. 1999 Feb 9;80(4):589-94. doi: 10.1002/(sici)1097-0215(19990209)80:4<589::aid-ijc17>3.0.co;2-d.
A majority of human breast carcinomas co-express the epidermal growth factor (EGF)-like peptides CRIPTO (CR), amphiregulin (AR) and transforming growth factor alpha (TGF-alpha). MDA-MB-468 breast carcinoma cells express CR, AR and TGFalpha, while SK-BR-3 cells express CR and TGF-alpha. Anti-sense phosphorothioate oligodeoxynucleotides (AS S-oligos) directed against either CR or TGF-alpha inhibit the proliferation of both cell lines. A 40-50% growth inhibition was observed at a 2-microM concentration of each AS S-oligo. Treatment with the AR AS S-oligo also resulted in a significant inhibition of MDA-MB-468 anchorage dependent growth (ADG). No significant growth inhibition was observed when MDA-MB-468 or SK-BR-3 cells were treated with a mis-sense S-oligo. The AS S-oligos inhibited the expression of AR, CR or TGF-alpha proteins and mRNAs, as assessed by immuno-cytochemistry and semi-quantitative RT-PCR. An additive growth-inhibitory effect was observed when MDA-MB-468 cells were treated with a combination of EGF-related AS S-oligos. Indeed, treatment of MDA-MB-468 cells with a combination of AR, CR and TGF-alpha AS S-oligos resulted in about 70% growth inhibition at a concentration of 0.7 microM each. Finally, treatment of MDA-MB-468 cells with a combination either of the 3 AS S-oligos or of an EGF receptor-blocking antibody (MAb 225) and either CR, AR or TGFalpha AS S-oligos resulted in a significant increase in DNA fragmentation. Our data suggest that the EGF-related peptides are involved in the proliferation and survival of breast carcinoma cells.
大多数人类乳腺癌共同表达表皮生长因子（EGF）样肽CRIPTO（CR）、双调蛋白（AR）和转化生长因子α（TGF-α）。MDA-MB-468乳腺癌细胞表达CR、AR和TGF-α，而SK-BR-3细胞表达CR和TGF-α。针对CR或TGF-α的反义硫代磷酸酯寡脱氧核苷酸（AS S-寡核苷酸）可抑制这两种细胞系的增殖。在每种AS S-寡核苷酸浓度为2μM时观察到40%-50%的生长抑制。用AR AS S-寡核苷酸处理也导致MDA-MB-468锚定依赖性生长（ADG）受到显著抑制。当用错义S-寡核苷酸处理MDA-MB-468或SK-BR-3细胞时，未观察到明显的生长抑制。通过免疫细胞化学和半定量RT-PCR评估，AS S-寡核苷酸抑制了AR、CR或TGF-α蛋白及mRNA的表达。当用与EGF相关的AS S-寡核苷酸组合处理MDA-MB-468细胞时，观察到了相加的生长抑制作用。实际上，用AR、CR和TGF-α AS S-寡核苷酸组合处理MDA-MB-468细胞，在每种浓度为0.7μM时导致约70%的生长抑制。最后，用3种AS S-寡核苷酸组合或用EGF受体阻断抗体（单克隆抗体225）与CR、AR或TGF-α AS S-寡核苷酸之一处理MDA-MB-468细胞，导致DNA片段化显著增加。我们的数据表明，与EGF相关的肽参与了乳腺癌细胞的增殖和存活。
