Crickard K, Gross J L, Crickard U, Yoonessi M, Lele S, Herblin W F, Eidsvoog K
Department of Gynecology and Obstetrics, SUNY at Buffalo, New York 14222.
Gynecol Oncol. 1994 Nov;55(2):277-84. doi: 10.1006/gyno.1994.1290.
Basic fibroblast growth factor (bFGF) and other members of the FGF family share several biological properties that have the potential to mediate neoplastic cell growth. To test the hypothesis that bFGF may play a role in human ovarian cancer cell growth, three ovarian cancer cell lines, A90, A121(P), and A121(A), were investigated for their ability to respond to bFGF as a mitogen, to express endogenous bFGF protein or message for FGF proteins, and to exhibit FGF receptor or its message. Addition of bFGF to cultures of all three cell lines maintained in chemically defined media resulted in a statistically significant increase in cell number. Cell extracts from A90, A121(P), and A121(A) contained an immunoreactive protein that comigrated with hr-bFGF by Western blot analysis. Several bands of higher molecular weight were also noted. Immunohistochemical staining for bFGF demonstrated a cytoplasmic distribution of bFGF in the three cell lines. Both high- and low-affinity binding sites for human recombinant bFGF (hr-bFGF) were expressed by all three lines. High-affinity sites varied from 2700 sites per cell (Kd = 29 pM) to 13,500 sites per cell (Kd = 71 pM). All three cell lines were screened for mRNA expression for seven FGF proteins and four FGF receptors. In all three lines, mRNA for FGF2 (bFGF) was detected by PCR analysis, and in two lines, mRNA for FGF1 (aFGF) and FGF5 were also found. The FGFR1 receptor subtype (flg) was common to all of the cell lines. Finally, suramin inhibited proliferation of A90 and A121 (P and A) with IC50's of 60 and 210 micrograms/ml, respectively. This is consistent with the A90 cell line having higher levels of endogenous bFGF and flg and therefore being more responsive to suramin inhibition than the A121 cell line. The results indicate that these ovarian cancer cell lines can produce bFGF as well as other members of the FGF family of genes and have the ability to respond to bFGF.
碱性成纤维细胞生长因子（bFGF）和FGF家族的其他成员具有多种生物学特性，有可能介导肿瘤细胞的生长。为了验证bFGF可能在人卵巢癌细胞生长中发挥作用这一假说，对三种卵巢癌细胞系A90、A121(P)和A121(A)进行了研究，考察它们作为有丝分裂原对bFGF作出反应的能力、表达内源性bFGF蛋白或FGF蛋白信息的能力，以及展示FGF受体或其信息的能力。向在化学成分明确的培养基中培养的所有三种细胞系的培养物中添加bFGF，导致细胞数量在统计学上显著增加。通过蛋白质印迹分析，A90、A121(P)和A121(A)的细胞提取物含有一种与hr-bFGF迁移率相同的免疫反应性蛋白。还注意到几条分子量更高的条带。bFGF的免疫组织化学染色显示bFGF在这三种细胞系中呈细胞质分布。所有三种细胞系均表达人重组bFGF（hr-bFGF）的高亲和力和低亲和力结合位点。高亲和力位点从每个细胞2700个位点（Kd = 29 pM）到每个细胞13500个位点（Kd = 71 pM）不等。对所有三种细胞系进行了七种FGF蛋白和四种FGF受体的mRNA表达筛选。在所有三种细胞系中，通过PCR分析检测到FGF2（bFGF）的mRNA，在两种细胞系中还发现了FGF1（aFGF）和FGF5的mRNA。FGFR1受体亚型（flg）在所有细胞系中都很常见。最后，苏拉明抑制A90和A121（P和A）的增殖，IC50分别为60和210微克/毫升。这与A90细胞系具有更高水平的内源性bFGF和flg一致，因此比A121细胞系对苏拉明抑制更敏感。结果表明，这些卵巢癌细胞系能够产生bFGF以及FGF家族的其他基因成员，并具有对bFGF作出反应的能力。
