The cytotoxic effect of Bowman-Birk isoinhibitors, IBB1 and IBBD2, from soybean (Glycine max) on HT29 human colorectal cancer cells is related to their intrinsic ability to inhibit serine proteases
The cytotoxic effect of Bowman-Birk isoinhibitors, IBB1 and IBBD2, from soybean (Glycine max) on HT29 human colorectal cancer cells is related to their intrinsic ability to inhibit serine proteases
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DOI：
10.1002/mnfr.200900122
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发表时间：
2010-03-01
影响因子：
5.2
通讯作者：
Domoney, Claire
中科院分区：
文献类型：
--
作者：
Clemente, Alfonso;Javier Moreno, Francisco;Domoney, Claire
Bowman比尔克抑制剂（BBI）是一种天然存在的蛋白酶抑制剂，在胃肠道内具有潜在的健康促进作用。在这项工作中，我们研究了大豆BBI蛋白对HT 29结肠腺癌细胞的影响，并与非恶性结肠成纤维细胞CCD-18 Co细胞进行了比较。两个主要的大豆异源抑制剂，IBB 1和IBBD 2，显示相当大的氨基酸序列的差异在其抑制结构域，纯化，以检查其功能特性，包括其对HT 29结肠癌细胞增殖的个别影响。IBB 1抑制胰蛋白酶和胰凝乳蛋白酶，而IBBD 2仅抑制胰蛋白酶。尽管IBB 1和IBBD 2的酶抑制特性显示出显著差异，但测定的IBB 1和IBBD 2对HT 29细胞生长的抑制浓度中值没有显著差异（分别为39.9 +/-2.3和48.3 +/-3.5 μ M）。HT 29结肠癌细胞的细胞周期分布模式受到BBI处理的剂量依赖性影响，细胞被阻滞在G 0-G1期。化学惰性的大豆BBI对HT 29细胞的增殖具有微弱但不显著的影响。大豆中BBI异源抑制剂的抗增殖特性表明，参与致癌作用的胰蛋白酶和胰凝乳蛋白酶样蛋白酶应被视为BBI样蛋白的潜在靶点。
Bowman Birk inhibitors (BBI) from soybean and related proteins are naturally occurring protease inhibitors with potential health-promoting properties within the gastrointestinal tract. In this work, we have investigated the effects of soybean BBI proteins on HT29 colon adenocarcinoma cells, compared with non-malignant colonic fibroblast CCD-18Co cells. Two major soybean isoinhibitors, IBB1 and IBBD2, showing considerable amino acid sequence divergence within their inhibitory domains, were purified in order to examine their functional properties, including their individual effects on the proliferation of HT29 colon cancer cells. IBB1 inhibited both trypsin and chymotrypsin whereas IBBD2 inhibited trypsin only. Despite showing significant differences in their enzyme inhibitory properties, the median inhibitory concentration values determined for IBB1 and IBBD2 on HT29 cell growth were not significantly different (39.9 +/- 2.3 and 48.3 +/- 3.5 mu M, respectively). The cell cycle distribution pattern of HT29 colon cancer cells was affected by BBI treatment in a dose-dependent manner, with cells becoming blocked in the G0-G1 phase. Chemically inactive soybean BBI had a weak but non-significant effect on the proliferation of HT29 cells. The anti-proliferative properties of BBI isoinhibitors from soybean reveal that both trypsin- and chymotrypsin-like proteases involved in carcinogenesis should be considered as potential targets of BBI-like proteins.