Ho T Y, Russo J, Russo I H
Department of Pathology, Fox Chase Cancer Center, Philadelphia, PA 19111.
Electrophoresis. 1994 May;15(5):746-50. doi: 10.1002/elps.11501501102.
Numerous attempts have made to describe the particular protein pattern of malignant cells by using high resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The placental hormone human chorionic gonadotropin (hCG) inhibits tumor initiation and progression in experimental animals and has an inhibitory effect on the proliferation of human breast epithelial cells (HBEC) in vitro. The inhibitory effect on the immortalized HBEC MCF-10F is accompanied by the immunocytochemical expression of inhibin alpha and beta subunits by treated cells. With the purpose of clarifying the molecular mechanisms involved in this effect, the pattern of protein synthesis and mRNA were studied by 2-D PAGE in the immortalized HBEC MCF-10F cells treated in vitro 1001U for 24 h. The effect of hCG treatment on the synthesis of MCF-10F cells was monitored by labeling both control and treated cells with [S35]methionine and separation by 2-D PAGE. At least 11 proteins were preferentially synthesized and five specific polypeptides were decreased in hCG treated cells in comparison with controls. The hCG induced at least four new mRNAs which encoded protein in the molecular mass range of 24-72 kDa. It also increased the expression of at least six mRNAs and reduced the expression of least four mRNAs in comparison with control cells. The hCG-treated cells actively synthesized a 33-kDa polypeptide which was not present in control cells. The nature of this hCG-inducible 33 kDa protein elucidated by immunoprecipating [S35]methionine-labeled proteins with antisera directed against rat inhibin subunit alpha and beta b.(ABSTRACT TRUNCATED AT 250 WORDS)
人们已多次尝试通过高分辨率二维聚丙烯酰胺凝胶电泳(2-D PAGE)来描述恶性细胞的特定蛋白质模式。胎盘激素人绒毛膜促性腺激素(hCG)在实验动物中可抑制肿瘤起始和进展,并且在体外对人乳腺上皮细胞(HBEC)的增殖具有抑制作用。对永生化的HBEC MCF-10F细胞的抑制作用伴随着处理后细胞中抑制素α和β亚基的免疫细胞化学表达。为了阐明这种作用所涉及的分子机制,通过2-D PAGE研究了在体外以1001U处理24小时的永生化HBEC MCF-10F细胞中的蛋白质合成模式和mRNA。通过用[S35]甲硫氨酸标记对照细胞和处理后的细胞并通过2-D PAGE进行分离,监测hCG处理对MCF-10F细胞合成的影响。与对照相比,hCG处理的细胞中至少有11种蛋白质被优先合成,并且有5种特定多肽减少。hCG诱导了至少4种新的mRNA,其编码分子量在24 - 72 kDa范围内的蛋白质。与对照细胞相比,它还增加了至少6种mRNA的表达,并降低了至少4种mRNA的表达。hCG处理的细胞积极合成了一种对照细胞中不存在的33 kDa多肽。通过用针对大鼠抑制素亚基α和βb的抗血清免疫沉淀[S35]甲硫氨酸标记的蛋白质,阐明了这种hCG诱导的33 kDa蛋白质的性质。(摘要截断于250字)