Nonhematopoietic tumor cell lines express stem cell factor and display c-kit receptors.
Nonhematopoietic tumor cell lines express stem cell factor and display c-kit receptors.
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DOI：
10.1182/blood.v80.2.374.bloodjournal802374
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发表时间：
1992-07
期刊：
影响因子：
20.3
通讯作者：
Anne M. Turner;K. Zsebo;FH Martin;Fredrick W. Jacobsen;Larry G. Bennett;V. C. Broudy
中科院分区：
文献类型：
--
作者：
Anne M. Turner;K. Zsebo;FH Martin;Fredrick W. Jacobsen;Larry G. Bennett;V. C. Broudy
人干细胞因子（SCF）在其他生长因子存在的情况下，刺激原始造血祖细胞的生长。这些作用是通过激活SCF受体c-kit来实现的。由于SCF在化疗和骨髓移植患者中的潜在应用，因此SCF对非造血肿瘤的影响需要进一步研究。为了确定人类肿瘤细胞系是否显示c-kit受体，我们在乳腺癌细胞系（Du4475）、胃癌细胞系（KATO III）、黑色素瘤细胞系（HTT144）以及两种小细胞肺癌细胞系（H69和H128）上进行了125I-SCF的结合实验。SCF对肿瘤细胞系的生物学效应是通过其刺激氚化胸腺嘧啶摄取和促进甲基纤维素中菌落生长的能力来评估的。乳腺癌细胞系Du4475和两种小细胞肺癌细胞系H69和H128表现出高亲和力的c-kit受体，其结合亲和力分别为40、100和90 pmol/L。每个细胞的高亲和受体数量从700到9500不等。胃癌细胞系和黑色素瘤细胞系均显示微量125I-SCF结合。SCF单独存在，或与粒细胞-巨噬细胞集落刺激因子或白细胞介素-3联合使用时，Du4475、H69或H128细胞系的集落生长增加不到17%。假设缺乏生长反应可能是肿瘤细胞系内源性SCF产生的继发因素，我们使用RNAse保护实验来确定肿瘤细胞系是否含有SCF信使RNA （mRNA）。此外，我们用酶免疫法检测肿瘤细胞系上清液中分泌的SCF蛋白的存在，并用间接免疫荧光法分析肿瘤细胞系中膜结合的SCF。我们的研究结果表明，Du4475、H69和H128细胞系以及黑色素瘤细胞系（HTT144）具有多个SCF mRNA拷贝。在Du4475和H69细胞系的细胞上清液中检测到可溶性SCF蛋白，并且在四种细胞系的表面都发现了SCF。这些数据表明，一些人实体瘤细胞系显示出高亲和力的c-kit受体，并产生可在细胞表面检测到的SCF。这些结果表明，携带c-kit受体的肿瘤细胞自分泌SCF可能会促进肿瘤细胞系的细胞生长。
Human stem cell factor (SCF) acts in the presence of other growth factors to stimulate the growth of primitive hematopoietic progenitor cells. These effects are performed by activation of the SCF receptor, c-kit. Because of the potential use of SCF in patients undergoing chemotherapy and bone marrow transplantation, the effect of SCF on nonhematopoietic tumors requires investigation. To determine whether human tumor cell lines display c-kit receptors, we performed binding experiments with 125I-SCF on a breast carcinoma cell line (Du4475), a gastric carcinoma cell line (KATO III), a melanoma cell line (HTT144), as well as two small cell lung carcinoma cell lines (H69 and H128). The biologic effect of SCF on tumor cell lines was assessed by its ability to stimulate tritiated thymidine uptake and to enhance colony growth in methylcellulose. The breast carcinoma cell line, Du4475, as well as two small cell lung carcinoma cell lines, H69 and H128, exhibit high-affinity c-kit receptors with approximate binding affinities of 40, 100, and 90 pmol/L, respectively. The number of high-affinity receptors per cell ranged from 700 to 9,500. The gastric carcinoma cell line, as well as the melanoma cell line, showed trace binding of 125I-SCF. In the presence of SCF alone, or in combination with granulocyte-macrophage colony-stimulating factor or interleukin-3, there was less than a 17% increase in the colony growth of Du4475, H69, or H128 cell lines. Postulating that the lack of growth response could be secondary to endogenous SCF production by the tumor cell lines, we used an RNAse protection assay to determine whether the tumor cell lines contain SCF messenger RNA (mRNA). In addition, we tested tumor cell line supernatants for the presence of secreted SCF protein by enzyme immunoassay, and analyzed the tumor cell lines for membrane-bound SCF by indirect immunofluorescence. Our results show that the Du4475, H69, and H128 cell lines, as well as a melanoma cell line (HTT144), have multiple copies of SCF mRNA. Soluble SCF protein was detected in the cell supernatants in the Du4475 and H69 cell lines and SCF was found on the surface of all four cell lines. These data show that some human solid tumor cell lines display high-affinity c-kit receptors and produce SCF, which can be detected on the cell surface. These results suggest the possibility that autocrine production of SCF by c-kit receptor-bearing tumor cells may enhance cell growth in tumor cell lines.