Evidence for autocrine growth stimulation of cultured colon tumor cells by a gastrin/cholecystokinin-like peptide.
Evidence for autocrine growth stimulation of cultured colon tumor cells by a gastrin/cholecystokinin-like peptide.
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胃泌素/胆囊收缩素样肽刺激培养的结肠肿瘤细胞自分泌生长的证据。
DOI：
10.1016/0014-4827(90)90204-n
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发表时间：
1990
影响因子：
3.7
通讯作者：
Brattain,MG
中科院分区：
文献类型：
--
作者：
Hoosein,NM;Kiener,PA;Curry,RC;Brattain,MG
肽胃泌素已被证明对正常以及恶性胃肠道组织具有生长刺激作用。在这项研究中，我们研究了自分泌生长刺激培养的结肠肿瘤细胞的胃泌素样肽的可能性。胃泌素/ CCK受体拮抗剂二丁酰cGMP以剂量依赖性方式抑制两种人结肠癌细胞系HCT 116（EC 50 = 1.3 mM）和CBS（EC 50 = 2.5 mM）的增殖。1 mM二丁酰cGMP作用于HCT 116细胞后，细胞形态发生明显改变。在受体结合试验中，二丁酰cGMP与125 I标记的胃泌素竞争结合HCT 116细胞（IC 50 = 1.8 mM）。环GMP的另一种衍生物8-溴cGMP由于其对胃泌素/CCK受体的亲和力相当弱而用作对照，不与放射性标记的胃泌素竞争结合HCT 116细胞，并且在浓度高达10 mM时对HCT 116或CBS细胞单层培养物中的形态或增殖没有影响。对适应在无血清培养基中生长的HCT 116和CBS细胞的依赖性细胞生长抑制作用。胃泌素/CCK受体拮抗剂和抗胃泌素/CCK抗体的抗增殖作用表明，这些细胞分泌的胃泌素样肽可能促进生长。放射免疫分析表明，这两种细胞的条件培养液中均含有胃泌素样肽（10-50 pg/106细胞/72 h）。使用与编码人胃泌素的十二肽羧基末端的核苷酸序列互补的寡核苷酸DNA探针的北方分析显示与探针杂交的三个转录物（0.7、3.3和3.7 kb）。这些数据首次为胃泌素/CCK样肽在培养的结肠肿瘤细胞中的自分泌生长刺激作用提供了证据。
The peptide gastrin has been shown to have growth stimulatory effects on normal as well as malignant gastrointestinal tissue. In this study, we have examined the possibility of autocrine growth-stimulation of cultured colon tumor cells by a gastrin-like peptide. The gastrin/ CCK receptor antagonist dibutyryl cGMP inhibited the proliferation of two human colon carcinoma cell lines HCT 116 (EC50= 1.3 mM) and CBS (EC50= 2.5 mM) in a dose-dependent manner. Marked morphological changes were observed in HCT 116 cells after treatment with 1 mMdibutyryl cGMP. In receptor binding assays, dibutyryl cGMP competed with125I-labeled gastrin for binding to HCT 116 cells (IC50= 1.8 mM). Another derivative of cyclic GMP, 8-Bromo cGMP used as control due to its considerably weaker affinity for the gastrin/CCK receptor, did not compete with radiolabeled gastrin for binding to HCT 116 cells and had no effect on the morphology or proliferation in monolayer cultures of HCT 116 or CBS cells at concentrations up to 10 mM. Antigastrin/CCK antisera was also found to have dose-dependent cytostatic effects on HCT 116 and CBS cells adapted to growth in serum-free medium. The antiproliferative effect of the gastrin/CCK receptor antagonist and antigastrin/CCK antibodies suggested that a gastrin-like peptide secreted by these cells may promote growth. Radioimmunoassay of the conditioned medium of these two cell lines indicated the presence of a gastrin-iike peptide (10–50 pg/106cells/72 h). Northern analysis using an oligonucleotide DNA probe complementary to the nucleotide sequence coding the dodecapeptide carboxyl terminal of human gastrin showed three transcripts (0.7, 3.3, and 3.7 kb) that hybridized with the probe. These data provide, for the first time, evidence for an autocrine growth stimulatory role of a gastrin/CCK-like peptide in cultured colon tumor Cells.