Zhang Ting, Chen Xiaolan, Qu Lingbo, Wu Jinglan, Cui Ran, Zhao Yufen
Key Laboratory of Chemical Biology, Department of Chemistry, Zhengzhou University, Zhengzhou 450052, PR China.
Bioorg Med Chem. 2004 Dec 1;12(23):6097-105. doi: 10.1016/j.bmc.2004.09.013.
To improve the biological activities of chrysin (CR), we synthesize Diethyl Chysin-7-yl phosphate (CPE: C(19)H(19)O(7)P) and tetraethyl bis-phosphoric ester of chrysin (CP: C(23)H(28)O(10)P(2)) through a simplified Atheron-Todd reaction. The interactions of the CR and CPE with lysozyme were explored by electrospray ionization mass spectrometry (ESI) and fluorescence spectrometry method. Experimental results indicate that CPE could form the noncovalent compound with lysozyme, while the interaction of the CR with lysozyme was not detected. In addition, whether and how the compounds CPE and CP affect proliferation and apoptosis in human cervical cancer Hela cells were investigated. Moreover, the effects of CPE and CP in Hela cells were compared with that of the nonmodified CR compound. The Hela cells were co-cultured with CR, CP, and CPE as experimental groups, respectively, and corresponding control groups treated without CR, CP, and CPE. The proliferation and apoptosis were detected using MTT assay, HCl denatured-methyl green-pyronin staining, PCNA immunohistochemistry and TUNEL techniques. The cell growth IC(50), relative absorbance (RA), proliferating index (PI), PCNA-IR (immunoreactivity IR) integration value (IV), and apoptosis index (AI) were calculated and their correlation was analyzed in each group. The results show that all CR, CP, and CPE could inhibit proliferation and induce apoptosis in Hela cells. Moreover, the effects of CP and CPE were more potent than that of CR. The CP and CPE were proved to be a kind of stronger apoptosis inducers than nonphosphated CR. There was a negative correlation between proliferation and apoptosis. In conclusion, the CR, CP, and CPE could effectively inhibit growth by down-regulated expression of PCNA, and induce apoptosis in Hela cells. The efficiency of the modified CP and CPE preceded nonmodified CR compounds. The CP and CPE may be a new potential anti-cancer drug for therapy of human cervical carcinoma.
为提高白杨素（CR）的生物活性，我们通过简化的阿特隆 - 托德反应合成了白杨素 -7- 磷酸二乙酯（CPE：C(19)H(19)O(7)P）和白杨素双磷酸四乙酯（CP：C(23)H(28)O(10)P(2)）。采用电喷雾电离质谱（ESI）和荧光光谱法探究了CR和CPE与溶菌酶的相互作用。实验结果表明，CPE能与溶菌酶形成非共价化合物，而未检测到CR与溶菌酶的相互作用。此外，研究了化合物CPE和CP是否以及如何影响人宫颈癌Hela细胞的增殖和凋亡。而且，将CPE和CP在Hela细胞中的作用与未修饰的CR化合物进行了比较。分别将Hela细胞与CR、CP和CPE共培养作为实验组，相应的对照组不进行CR、CP和CPE处理。使用MTT法、盐酸变性 - 甲基绿 - 派洛宁染色、PCNA免疫组织化学和TUNEL技术检测增殖和凋亡情况。计算每组的细胞生长IC(50)、相对吸光度（RA）、增殖指数（PI）、PCNA - IR（免疫反应性IR）积分值（IV）和凋亡指数（AI），并分析它们之间的相关性。结果表明，所有的CR、CP和CPE均能抑制Hela细胞增殖并诱导其凋亡。而且，CP和CPE的作用比CR更强。CP和CPE被证明是比未磷酸化的CR更强的凋亡诱导剂。增殖与凋亡之间呈负相关。总之，CR、CP和CPE可通过下调PCNA表达有效抑制生长，并诱导Hela细胞凋亡。修饰后的CP和CPE的效果优于未修饰的CR化合物。CP和CPE可能是治疗人宫颈癌的新型潜在抗癌药物。
