Iizuka Norio, Oka Masaaki, Yamamoto Kotaro, Tangoku Akira, Miyamoto Koji, Miyamoto Takanobu, Uchimura Shunji, Hamamoto Yoshihiko, Okita Kiwamu
Department of Bioregulatory Function, Yamaguchi University School of Medicine, Yamaguchi, Japan.
Int J Cancer. 2003 Nov 20;107(4):666-72. doi: 10.1002/ijc.11452.
Although the physiological actions of many herbs are gradually being elucidated at the molecular level, it remains unclear how individual components of herbs contribute to their biological activities. In the present study, the antiproliferative activity of Coptidis rhizoma, a medicinal herb, and the major component berberine was investigated in 8 human pancreatic cancer cell lines. Gene expression patterns associated with sensitivities to each agent were analyzed with oligonucleotide arrays that comprised approximately 11,000 genes. We used a tetrazolium dye (MTT) assay to determine ID(50) values after the 8 cell lines were exposed to the 2 agents for 72 hr. The ID(50) value for berberine was correlated positively with that for C. rhizoma (r=0.725, p=0.0401). C. rhizoma killed tumor cells more effectively than purified berberine when normalized to the level of berberine present in the herb. From the oligonucleotide array data, we selected 20 and 13 genes with strong correlations (r(2)>0.81) to ID(50) values for berberine and C. rhizoma, respectively. Among these 33 genes, the levels of expression of 12 were correlated with the ID(50) values of both agents, suggesting that these genes are associated with tumor-killing activity of berberine in C. rhizoma. Expression of the remaining 21 genes was correlated with the ID(50) value of either purified berberine or C. rhizoma. Thus, we identified common and distinct genes responsible for anti-proliferative activities of purified berberine and C. rhizoma. This strategy may improve our understanding of the actions of herbs with antitumor activities.
尽管许多草药的生理作用在分子水平上正逐渐被阐明，但草药的各个成分如何对其生物活性产生影响仍不清楚。在本研究中，我们对一种草药黄连及其主要成分黄连素在8种人胰腺癌细胞系中的抗增殖活性进行了研究。使用包含约11000个基因的寡核苷酸阵列分析了与对每种药物敏感性相关的基因表达模式。在8种细胞系暴露于这两种药物72小时后，我们使用四唑盐染料（MTT）测定法来确定半数抑制浓度（ID50）值。黄连素的ID50值与黄连的ID50值呈正相关（r = 0.725，p = 0.0401）。相对于草药中黄连素的含量进行标准化后，黄连比纯化的黄连素更有效地杀死肿瘤细胞。从寡核苷酸阵列数据中，我们分别选择了与黄连素和黄连的ID50值具有强相关性（r2>0.81）的20个和13个基因。在这33个基因中，12个基因的表达水平与两种药物的ID50值相关，表明这些基因与黄连中黄连素的肿瘤杀伤活性有关。其余21个基因的表达与纯化的黄连素或黄连的ID50值相关。因此，我们确定了负责纯化黄连素和黄连抗增殖活性的共同和不同基因。这种策略可能会增进我们对具有抗肿瘤活性草药作用的理解。
