胃泌素及其受体拮抗剂对BGC—823细胞系生长的调节

本文用液闪测定和细胞计数观察了胃泌素及其受体拮抗剂丙谷胺和Ｌ－３６５２６０对体外培养的人胃腺癌ＢＧＣ－８２３细胞系的生长调节作用，结果表明不同浓度胃泌素对ＢＧＣ－８２３细胞系的生长均有显著促进作用，而这种作用可被其受体拮抗剂丙谷胺和Ｌ－３６５２６０所抑制，且发现在胃泌素浓度衡定时，Ｌ－３６５２６０对ＢＧＣ－８２３细胞系的生长抑制作用明显强于丙谷胺。这一结果也提示，Ｌ－３６５２６０对胃泌素受体的亲合     

促胃液素及其受体拮抗剂丙谷胺对人胃癌细胞系生长的影响

目的观察和分析五肽促胃液素ＰＧ及其受体拮抗剂丙谷胺ＰＧＭ对人胃癌细胞系ＭＧＣ生长的影响,为临床应用促胃液素受体拮抗剂协助治疗胃癌提供依据．方法选用５ｍｇ／Ｌ,１０ｍｇ／Ｌ,１５ｍｇ／Ｌ,２０ｍｇ／Ｌ４种浓度的ＰＧ和３０ｍｇ／Ｌ的ＰＧＭ分别作用于体外培养的浓度为２５×１０８／Ｌ的ＭＧＣ,分别培养２４,４８,７２ｈ,于酶标仪上选用波长５４０ｎｍ测定吸光值Ａ,并对数据进行比较分析．结果４种浓度的ＰＧ作用于ＭＧＣ,ＭＧＣ连续３ｄ的生长状态与对照组无明显差异,而ＭＧＣ在ＰＧＭ作用下,其连续３ｄ的平均Ａ值分别为００２９,００４６和００８４,而未被ＰＧＭ作用的ＭＧＣ的平均Ａ值分别是０１０１,０１１５和０１８２,ＭＧＣ在ＰＧＭ作用下生长明显低于对照组（Ｐ＜００５）．结论外源性的ＰＧ对ＭＧＣ无营养促进作用,而ＰＧＭ能抑制ＭＧＣ的生长     

胃泌素及其受体拮抗剂丙谷胺对肝癌细胞株生长的影响

目的: 研究胃泌素及胃泌素受体拮抗剂丙谷胺对肝癌细胞株的增殖的影响,并探讨肝癌患者非细胞毒内分泌治疗的可能性.方法:采用MTT比色分析法研究在不同浓度胃泌素和其受体拮抗剂丙谷胺的干预下, 4种人肝癌细胞株的增殖情况, 选取1种细胞株, 用流式细胞仪分析在胃泌素及丙谷胺作用下的周期分布.结果: 胃泌素在接近生理浓度下促进QGY-7701和Bel-7402的增殖, 丙谷胺对胃泌素的作用有抑制作用. 过量的胃泌素对QGY-7701有抑制效果. 胃泌素对HepG2和SMMC-7721无明显促增殖作用. 丙谷胺在高浓度下对所有细胞株皆有明显的抑制作用. 胃泌素可促使Bel-7402 G0/G1期细胞向S、G2/M期转化, 加用丙谷胺后胃泌素的上述作用消失.结论:胃泌素对肝癌细胞株有促增殖作用及促进DNA合成, 丙谷胺对其有抑制作用, 可能成为肝癌内分泌辅助治疗的一个新的途径.     

胃泌素及生长抑素对人大肠癌细胞的调节

目的观察五肽胃泌素（ＰＧ）、胃泌素受体拮抗剂丙谷胺（ＰＧｌ）和生长抑素（ＳＳ）长效类似物ＳＭＳ２０１＿９９５对体外培养人大肠癌细胞系ＳＷ１１１６生长的影响。方法设空白组、对照组、ＰＧ组、ＰＧｌ组、ＳＳ组、ＰＧｌ＋ＰＧ组和ＳＳ＋ＰＧ组。各组主要药物浓度分５×１０－５＿５×１０－１０ｍｏｌ／Ｌ６个浓度，联合用药组ＰＧ浓度均为５×１０－５ｍｏｌ／Ｌ。采用ＭＴＴ比色分析法检测细胞增殖情况。结果ＰＧ能提高细胞增殖率；ＰＧｌ则抑制细胞增殖；ＰＧ对ＰＧｌ的抑制率总体影响不大，而使其与ＰＧｌ浓度对数值呈显著负相关（ｒ＝－０．８３，Ｐ＜０．０５）。ＳＳ在低浓度时促进细胞增殖，而高浓度时则有抑制作用，细胞增殖率与药物浓度对数值呈显著负相关（ｒ＝－０．９１，Ｐ＜０．０５）；ＰＧ可逆转ＳＳ的促增殖作用，并使其抑制率有所提高，且细胞增殖率仍与ＳＳ浓度对数值呈显著负相关（ｒ＝－０．８４，Ｐ＜０．０５）。结论激素可控制大肠癌的生长，但尚有许多问题待阐明     

胃泌素对结肠癌细胞株HT-29水通道蛋白4表达的影响

目的: 观察胃泌素对人结肠癌细胞株HT-29水通道蛋白4表达的影响,探讨胃泌素促进结肠癌侵润转移的另一可能机制.方法: 体外培养结肠癌细胞株,采用不同浓度的胃泌素(10-6、10-7、10-8 mol/L)干预HT-29细胞12 h,同时用10 mmol/L的胃泌素受体拮抗剂丙谷胺处理细胞1 h,再给予10-7 mol/L的胃泌素干预相同时间.采用免疫细胞化学和流式细胞技术的方法检测水通道蛋白4表达的变化.结果: 各浓度胃泌素干预后,细胞水通道蛋白4的表达与空白组和丙谷胺处理组相比均明显增加(16.08%±1.93%,17.00%±2.72%,16.48%±2.22% vs 9.28%±2.74%,8.52%±2.72%,均P＜0.01);丙谷胺处理组的表达与空白组无差别,胃泌素各浓度组间亦无差别.结论: 胃泌素能够增加结肠癌细胞水通道蛋白4的表达,丙谷胺能够阻断胃泌素的作用.     

胃泌素和丙谷胺对二甲基肼诱发大鼠大肠癌的影响

胃泌素和丙谷胺对二甲基肼诱发大鼠大肠癌的影响智发朝，李建国，张万岱，张振书，周殿元大肠癌的发生受许多因素影响，其中胃泌素及其受体拮抗剂─丙谷胺与大肠癌的发生和生长有密切关系［１］。材料与方法一、实验动物的分组和处理雄性Ｗｉｓｔａｒ大鼠１００只，体重１...     

血小板活化因子介导过氧化氢诱导肾小球系膜细胞-白细胞粘附

目的：研究过氧化氢（Ｈ２Ｏ２）对肾小球系膜细胞－中性粒细胞（ＧＭＣ－ＰＭＮ）粘附的影响及其机制。方法：用体外培养的人肾小球系膜细胞与不同浓度血小板活化因子（ＰＡＦ）或Ｈ２Ｏ２作用，并以各类拮抗剂观察其对ＧＭＣ－ＰＭＮ粘附的影响。结果：不同浓度Ｈ２Ｏ２促进ＧＭＣ依赖性ＰＭＮ粘附，以１０－２ｍｏｌ／Ｌ浓度的作用最强，使ＰＭＮ粘附率增强２．２倍。用ＰＡＦ受体拮抗剂预处理ＰＭＮ对粘附无影响。预处理ＧＭＣ可显著降低粘附率，磷酯酶Ａ２抑制剂对溴基苯酰基溴、钙调蛋白抑制剂氯丙嗪和钙离子螯合剂ＥＧＴＡ预处理ＧＭＣ，均能降低Ｈ２Ｏ２引起的ＧＭＣ－ＰＭＮ粘附。结论：Ｈ２Ｏ２可促进ＧＭＣ与ＰＭＮ的粘附。Ｈ２Ｏ２的这一作用可能是通过ＰＡＦ介导的。     

血管紧张素Ⅱ及其受体拮抗剂在系膜细胞培养中对结缔组织生长因子表达的影

目的：研究血管紧张素Ⅱ（ＡｎｇⅡ）及其受体拮抗剂Ｌｏｓａｒｔａｎ对系膜细胞（ＭＣｓ）结缔组织生长因子（ＣＴＧＦ）表达的影响。方法：分离培养ＳＤ大鼠肾小球系膜细胞。培养的系膜细胞中分别加入不同浓度的ＡｎｇⅡ（１０＾－９、１０＾－７、１０＾－５ｍｏｌ／Ｌ）,及１０＾－７ｍｏｌ／Ｌ ＡｎｇⅡ＋１０＾－５ｍｏｌ／Ｌ Ｌｏｓａｒｔａｎ,作用７２ｈ后,采用反转录－聚合酶链反应（ＲＴ－ＰＣＲ）技术测定ＭＣｓ ＣＴＧＦ ｍＲＮＡ水平变化。结果：ＡｎｇⅡ能促进ＭＣｓ ＣＴＧＦ ｍＲＮＡ表达,且呈剂量依赖性;Ｌｏｓａｒｔａｎ能部分降低ＡｎｇⅡ对ＣＴＧＦ ｍＲＮＡ表达的诱导。结论：ＡｎｇⅡ可能通过促进ＭＣｓ ＣＴＧＦ的表达而促进了肾纤维化的进展;Ｌｏｓａｒｔａｎ可以部分抑制ＡｎｇⅡ对ＣＴＧＦ ｍＲＮＡ表达的诱导,从而可能有益于延缓肾     

胃泌素受体拮抗剂丙谷胺对原代培养大肠癌细胞作用的研究

目的：在体外观察胃泌素受体持抗剂丙谷胺（PGL）对原代培养大肠癌活细胞数（吸光度，A值）和DNA合成（脉冲数，CPM值）的影响，为大肠癌病人临床应用胃泌素受体拮抗剂治疗提供实验依据。方法：对25例大肠癌根治术病人的新鲜标本进行细胞分离和原代培养，采用MTT比色分析法检测活细胞数；3H－TdR掺入法测定DNA合成．结果：PGL组高、中和低分化腺癌活细胞数和DNA合成与对照组比差异均无显著性（P均>0.05),5肽胃泌素（PG）组活细胞数和DNA合成均显著高于对照组（P均＜0．01），PGL＋PG组活细胞数和DNA合成均显著低于PG组（P均＜0．01），而与对照组比差异无显著性（P均＞0．05）。结论：丙谷胺对原代培养大肠癌细胞的增殖无明显影响，但可抑制胃泌素对大肠癌细胞的促增殖作用。     

TGF—β1表达与小鼠病毒性心肌炎细胞凋亡的关系

本实验通过给ＢＡＬＢ／Ｃ小鼠腹腔接种柯萨奇Ｂ－３ｍ病毒（ＣＶＢ３ｍ）诱发病毒性心肌炎（ＶＭＣ）感染病毒９ｄ后，ＶＭＣ检出率９３．３３％，应用原位末端标记法（Ｔ／ＶＮＥＬ）及免疫组化技术检测发现，７６．６７％感染病毒鼠心肌中发现凋亡细胞，７３．３３％，有ＴＧＦ－β１的表达，二者之间分布区域一致，提示细胞凋亡可能参与ＣＶＢ３ｍ诱导的ＶＭＣ的发生，发展，而ＴＧＦ－β１可能参与ＶＭＣ细胞凋亡的调控。     

Growth of colorectal carcinoma cells: regulation in vitro by gastrin,pentagastrin and the gastrin-receptor antagonist proglumide

Summary The growth-regulating effects of pentagastrin, gastrin and the gastrin-receptor antagonist proglumide were investigated in three established cell lines derived from human colorectal carcinomas in vitro and after transplantation into nude mice. In vitro a significant increase of cell growth in the SW 403 cell line incubated with pentagastrin or gastrin was observed. In the Lovo cell line this effect was only detected after synchronization of cell growth. Pentagastrin and gastrin had no effect on the growth of the Ls 174 T cell line. Proglumide reduced cell proliferation in all three cell lines as well as in the L929S cell line derived from fibroblasts, which served as control. After transplantation into nude mice all tumor cell lines increased, Love and Ls 174 T as undifferentiated tumor, SW 403 as differentiated. Pentagastrin increased and proglumide decreased growth in SW 403 tumors, whereas no effect was observed on Ls 174 T and Lovo tumors. We therefore conclude that growth of some colorectal carcinomas is regulated by gastrin, but that the effect of proglumide is unspecific rather than related to blockage of gastrin receptors. The growth-regulating effect of gastrin could be due to tumor differentiation.Abbreviations CEA carcinoembryonic antigen Supported by Deutsche Forschungsgemeinschaft (DFG EG 1/1-88)     

Effects of gastrin, proglumide, and somatostatin on growth of human colon cancer

The effects of gastrin, proglumide (a gastrin receptor antagonist), and somatostatin on growth of human colon adenocarcinoma cell lines CX1, X56, and HT29 were examined in two experimental models. Nude mice bearing xenografts of colon cancer CX1 or X56 were treated for 14-25 days subcutaneously with saline, pentagastrin (0.5 or 1.0 mg/kg), proglumide (250 or 500 mg/kg), or somatostatin 14 (33, 100, or 300 micrograms/kg) twice daily. Tumor volume, weight, protein, and deoxyribonucleic acid were measured. HT29 cells were grown in vitro and the effects of gastrin 17, proglumide, and somatostatin on growth were evaluated by cell counts or [3H]thymidine incorporation. The larger dose of pentagastrin significantly increased tumor growth in the nude mouse (p less than 0.005) and gastrin induced a biphasic effect on deoxyribonucleic acid synthesis in tissue culture with significant increases of up to 39% (p less than 0.025). Somatostatin alone significantly inhibited tumor growth in two of the cell lines and also inhibited the gastrin-induced growth. Proglumide had no effect by itself but significantly inhibited gastrin-stimulated growth. These findings suggest that growth of some human colon cancers may be hormone-dependent.     

Antiproliferative gastrin/cholecystokinin receptor antagonists target the 78-kDa gastrin-binding protein.

Inhibition of colon carcinoma cell growth by the nonselective gastrin/cholecystokinin (CCK) receptor antagonists proglumide and benzotript provided evidence that gastrin functions as an autocrine growth factor. However, the molecular properties of the receptor mediating the antagonist effects have not been identified. A 78-kDa gastrin-binding protein (GBP), the sequence of which is related to the family of enzymes possessing enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase activities, has been previously purified from porcine gastric mucosal membranes. I now report that covalent cross-linking of 125I-labeled [Nle15]gastrin2,17 to the 78-kDa GBP is inhibited by crotonyl-CoA and by acetoacetyl-CoA. Gastrin, CCK, and their analogues also inhibit cross-linking, and the spectrum of analogue affinities correlates better with the values previously reported for binding to the gastrin/CCK-C receptor than with the values reported for binding to either the CCK-A or the gastrin/CCK-B receptor. Cross-linking is also inhibited by proglumide and benzotript, but no inhibition is seen with either the CCK-A receptor-selective antagonist L364,718 or the gastrin/CCK-B receptor-selective antagonist L365,260. The affinities of antagonists for the GBP correlate well with their affinities for the gastrin/CCK-C receptor and with their potencies for inhibition of colon carcinoma cell growth. I conclude that the 78-kDa gastrin-binding protein is (i) a member of the hydratase/dehydrogenase family of fatty acid oxidation enzymes, (ii) the gastrin/CCK-C receptor, and (iii) the target for the antiproliferative action of two gastrin/CCK receptor antagonists.     

促胃液素对人结肠癌细胞株的生长调控及受体后信息传导

目的研究促胃液素对人结肠癌细胞株的生长调控及受体后的信息传导途径.方法以不同浓度的促胃液素处理人低分化结肠癌细胞株CloneA细胞,观察细胞的生长情况;先分别提取细胞内三磷酸肌醇(IP3)及环磷酸腺苷(cAMP),分别应用液体闪烁测量法、γ-闪烁记数仪测定其含量;用Fura-2负载及荧光测定细胞内游离Ca2+浓度([Ca2+]i).结果不同浓度的促胃液素10-9mol/L～10-5mol/L能明显地促进CloneA细胞的生长,光密度值分别为0.588±0.038,0.706±0.036,0.767±0.006,0.892±0.010,1.136±0.021;对照组为0.498±0.049(P＜0.01),且呈剂量依赖关系,这种促生长作用可以被促胃液素受体拮抗剂丙谷胺所阻断.促胃液素能显著促进CloneA细胞内IP3,Ca2+及cAMP含量增加,这些作用也可以被丙谷胺所抑制.结论促胃液素促进CloneA细胞生长,是由细胞膜上的促胃液素受体所介导,可能是通过磷酸肌醇和腺苷环化酶途径.     

Effect of gastrin and enprostil, a PGE2 analog, on colonic cancerous cell growth

The effects of gastrin (G-17), proglumide (a gastrin receptor antagonist), and enprostil (a synthetic analog of prostaglandin E2) used alone or in association were studied in colonic cancer Prob and Regb cell growth. The Prob (progressive in BD IX rats) and Regb (regressive) cell lines were cloned from a single chemically-induced rat colonic cancer. After a serum-free period corresponding to one doubling cell time, cells were incubated with 100 to 1,200 pM G-17, 40 or 80 mM proglumide, and 2.5 to 5 micrograms/ml enprostil for 8 h. Cell growth was measured 48 h later by colorimetric MTT assay. Two and four hundred pM G-17 gave a growth stimulation of 17.4 percent and 31 percent for Prob cells respectively or 35.5 percent and 49 percent for Regb cells. Growth stimulation was found to be statistically different (P less than 0.01) for Prob and Regb cells. Proglumide partially inhibited this growth stimulation whereas enprostil inhibited in totally. These results suggest that growth of some colonic cancer cell lines may be G-17 dependent. However the intensity of cell-growth stimulation depends on the level of cell malignancy or differentiation in a single tumor.     

Selective CCK-A but not CCK-B receptor antagonists inhibit HT-29 cell proliferation: synergism with pharmacological levels of melatonin

Some data suggest that cholecystokinin (CCK) receptor agonists stimulate the growth of colon cancer. Melatonin, an endogenous indoleamine with strong antioxidant properties, displays antiproliferative and proapoptotic properties both in vivo or in vitro in several types of tumors. We used HT-29 human colon cancer cells, expressing CCK receptors, to test the antiproliferative effects of several antagonists of CCK-A and/or CCK-B and their possible synergism with melatonin. HT-29 cells were cultured in RPMI 1640 medium supplemented with fetal bovine serum at 37 degrees C. Cell proliferation was assessed by the incorporation of [3H]-thymidine into DNA. Annexin V-FITC plus propidium iodine were used for flow cytometry apoptosis/necrosis evaluation. The following drugs were tested: gastrin (CCK-B agonist); CCK-8s (CCK-A agonist); proglumide (CCK-A plus CCK-B antagonist); lorglumide (CCK-A antagonist); PD 135,158 (CCK-B antagonist and weak CCK-A agonist); devazepide or L 364,718 (CCK-A antagonist); L 365,260 (CCK-B antagonist), and melatonin. The results shown a lack of effects of gastrin on HT-29 cell proliferation, whereas CCK-8s induced proliferation at high doses. The order of the antiproliferative effect of the other drugs was devazepide > lorglumide > proglumide. These drugs produce cell death mainly inducing apoptosis. Melatonin showed strong antiproliferative effect at millimolar concentrations, and it induced apoptotic cell death. Melatonin generally enhanced the antiproliferative effects of devazepide, lorglumide and proglumide and increased the proglumide-induced apoptosis. These results suggest that melatonin and CCK-A antagonists are useful for controlling human colon cancer cell growth in culture and in combined therapy significantly increases their efficiency.     

Regulatory effect and mechanism of gastrin and its antagonists on colorectal carcinoma

lNTRODUCTIONInrecentyears,s0mestudiesindicatedthatgastrincouldpromotethegrowthofsomecolorectalcarcinomas,butgastrinantag0nistsuchasPGL,somat0statin(SS)c0uldinhibitthegrowthofthosecolorectalcarcin0mas.Inordertoexplorethemechanismoftheeffectandtheclinicalsignificanceofgastrinanditsantagonistsoncolorectalcarcin0ma,weestablishedthemodeloftransplantedcol0niccarcin0mafromSW48Ocelllineingymnomousebody,andobservedthevolumeandweightoftransplantedcarcinoma,contentofcAMP,DNA,proteinandcellcycleo…     

Expression of progastrin-derived peptides and gastrin receptors in a panel of gastrointestinal carcinoma cell lines

To assess the potential of gastrin receptor antagonists in the treatment of gastrointestinal cancer, the presence of an autocrine loop involving progastrin-derived peptides has been investigated in two colorectal and one gastric carcinoma cell lines. Progastrin, glycine-extended gastrin and amidated gastrin were detected in cell extracts or conditioned media by radio-immunoassay. Low-affinity binding sites for glycine-extended gastrin and amidated gastrin were present, but high-affinity binding sites were not detected with the appropriate iodinated ligands. In addition, neither glycine-extended gastrin nor amidated gastrin in the concentration range 10pmol/L-10nmol/L stimulated cell proliferation. We conclude that it is unlikely that the carcinoma cell lines LIM 1215, LIM 1839 and LIM 1899 use either amidated or glycine-extended gastrins as extracellular autocrine growth factors.     

The role of gastrin and cholecystokinin in normal and neoplastic gastrointestinal growth

Abstract Gastrin and cholecystokinin (CCK) act as growth factors for the gastric mucosa and the pancreas, respectively. CCK is also responsible, via the CCK-A receptor, for the pancreatic hyperplasia observed following the feeding of protease inhibitors or pancreaticobiliary diversion. Hypergastrinaemia does not increase the incidence of spontaneous gastrointestinal carcinoma, but does stimulate the proliferation of gastric enterochromaffin-like cells via the gastrin/CCK-B receptor, with a consequent increase in the incidence of gastric carcinoids. Whether gastrin influences mutagen-induced gastrointestinal carcinogenesis is still controversial, but CCK clearly enhances the induction by carcinogens of acinar tumours in the pancreas. While gastrin increases xenograft growth of 50% of gastrointestinal tumours tested, effects on the proliferation of gastrointestinal tumour cell lines in vitro have been more difficult to demonstrate, perhaps because many cell lines are already maximally stimulated by autocrine gastrin. Gastrin mRNA and progastrin, but not mature amidated gastrin, have been detected in all gastrointestinal cell lines tested. Although cell proliferation is inhibited by gastrin/CCK receptor antagonists, the spectrum of antagonist affinities is not consistent with binding to either CCK-A or gastrin/CCK-B receptors. Definition of the molecular structure of the receptor involved in the autocrine loop may lead to novel therapies for gastrointestinal cancer.