Rao C V, Wang C Q, Simi B, Rodriguez J G, Cooma I, El-Bayoumy K, Reddy B S
Chemoprevention and Nutritional Carcinogenesis Program, American Health Foundation, Valhalla, New York 10595, USA.
Cancer Res. 2001 May 1;61(9):3647-52.
We have consistently shown that several synthetic Organoselenium compounds are superior cancer chemopreventive agents and less toxic than selenite or certain naturally occurring selenoamino acids. 1,4-Phenylenebis(methylene)selenocyanate (p-XSC) is the lead Organoselenium compound in that it has been shown to be the most effective and the least toxic agent in several experimental cancer models. It is not known whether p-XSC or one of its metabolites is responsible for its chemopreventive efficacy. As an initial step, we synthesized one of its putative metabolites, i.e., the glutathione conjugate of p-XSC (p-XSe-SG), and determined its stability in the pH range from 2 to 8 and in the diet under normal feeding conditions. We also assessed its maximum tolerated dose and examined its chemopreventive efficacy against azoxymethane (AOM)-induced colon carcinogenesis in male F344 rats. p-XSe-SG proved to be very stable over the pH range tested. The maximum tolerated dose of p-XSe-SG determined in a 6-week subchronic toxicity study was found to be >210 ppm (>40 ppm selenium) when the compound was added to AIN-76A high-fat diet. To assess the efficacy of this agent in the postinitiation period of colon carcinogenesis, male F344 rats 6 weeks of age were fed the high-fat diet, and at beginning of weeks 7 and 8, all of the rats intended for carcinogen treatment were given AOM at a dose of 15 mg/kg body weight by s.c. injection. Two days after the carcinogen treatment, the groups of rats consuming the high-fat control diet began their respective high-fat experimental diet regimens with 0, 56, or 84 ppm p-XSe-SG (0.1, 10, and 15 ppm of selenium) supplementation. All animals continued on their respective diets for 38 weeks after the AOM-treatment and were then killed. Colon tumors were evaluated histologically using routine procedures and were also analyzed for cyclooxygenase (COX)-1 and COX-2 expression and enzymatic activities. The results indicate that p-XSeSG administered during the post-initiation stage significantly inhibited both the incidence (P < 0.05-0.01) and the multiplicity (P < 0.05-0.005) of AOM-induced colon adenocarcinomas. This agent also greatly suppressed the multiplicity (P < 0.01-0.001) of AOM-induced exophytic adenocarcinomas in a dose-dependent manner. Feeding of 56 or 84 ppm p-XSe-SG in the diet significantly suppressed total COX activity (P < 0.02 to -0.01) and COX-2 specific activity (P < 0.005-0.0005) but had minimal effect on the protein expression levels of COX-1 and COX-2. These results suggest that the newly developed synthetic Organoselenium compound, p-XSe-SG, is stable in the diet and at wide pH ranges, inhibits colon carcinogenesis when administered during the postinitiation stage, and inhibits COX activity. Compared with previous efficacy studies and considering the toxicity associated with selenium, p-XSe-SG seems to be the least toxic Organoselenium chemopreventive agent thus far tested in the experimental colon carcinogenesis. Studies are in progress to delineate whether p-XSe-SG is also effective when administered during the progression stage of colon carcinogenesis.
我们一直表明,几种合成有机硒化合物是优越的癌症化学预防剂,且比亚硒酸盐或某些天然存在的硒代氨基酸毒性更低。1,4-亚苯基双(亚甲基)硒氰酸盐(p-XSC)是主要的有机硒化合物,因为在多个实验性癌症模型中它已被证明是最有效且毒性最小的试剂。尚不清楚是p-XSC还是其代谢产物之一导致了它的化学预防功效。作为第一步,我们合成了其一种假定的代谢产物,即p-XSC的谷胱甘肽共轭物(p-XSe-SG),并确定了它在pH值2至8范围内以及正常喂养条件下在饮食中的稳定性。我们还评估了它的最大耐受剂量,并检测了它对雄性F344大鼠中由氧化偶氮甲烷(AOM)诱导的结肠癌发生的化学预防功效。结果证明p-XSe-SG在所测试的pH范围内非常稳定。在一项为期6周的亚慢性毒性研究中,当该化合物添加到AIN-76A高脂饮食中时,所确定的p-XSe-SG的最大耐受剂量>210 ppm(>40 ppm硒)。为了评估该试剂在结肠癌发生起始后阶段的功效,给6周龄的雄性F344大鼠喂食高脂饮食,在第7周和第8周开始时,所有打算接受致癌物处理的大鼠通过皮下注射给予剂量为15 mg/kg体重的AOM。致癌物处理两天后,食用高脂对照饮食的大鼠组开始各自的高脂实验饮食方案,分别补充0、56或84 ppm的p-XSe-SG(0.1、10和15 ppm的硒)。在AOM处理后,所有动物继续各自的饮食38周,然后处死。使用常规程序对结肠肿瘤进行组织学评估,并分析其环氧合酶(COX)-1和COX-2的表达及酶活性。结果表明,在起始后阶段给予p-XSeSG可显著抑制AOM诱导的结肠腺癌的发生率(P < 0.05 - 0.01)和多发性(P < 0.05 - 0.005)。该试剂还以剂量依赖的方式极大地抑制了AOM诱导的外生性腺癌的多发性(P < 0.01 - 0.001)。饮食中添加56或84 ppm的p-XSe-SG可显著抑制总COX活性(P < 0.02至 -0.01)和COX-2的比活性(P < 0.005 - 0.0005),但对COX-1和COX-2的蛋白表达水平影响最小。这些结果表明,新开发的合成有机硒化合物p-XSe-SG在饮食和较宽的pH范围内稳定,在起始后阶段给予时可抑制结肠癌发生,并抑制COX活性。与先前的功效研究相比,并考虑到与硒相关的毒性,p-XSe-SG似乎是迄今为止在实验性结肠癌发生中测试的毒性最小的有机硒化学预防剂。正在进行研究以确定p-XSe-SG在结肠癌发生进展阶段给予时是否也有效。