抗肿瘤化合物CENPOANU的体外抗肿瘤活性研究

目的：研究抗肿瘤化合物1-（2-氯乙基）．3．2．（2-硝基苯氧）乙酰基]-1-亚硝基脲（CENPOANU）的体外抗肿瘤活性及其机制。方法：考察对人乳腺癌细胞（McF-7）、人宫颈癌细胞（HeLa）、小鼠红白血病细胞（MEL）、人恶性胶质瘤细胞（u251），CEN—POANU和卡莫司汀（BCNU）的半数抑菌浓度（IC。。）；检测0、0．5、1．O、1．5、2．O、4．O倍IC。的CENPOANU作用24、36、48h后MCF．7的增殖抑制率，0、0．5、1．0、1．5、2．O、4．0倍IC50的CENPOANU、BCNU作用48h后MCF．7的增殖抑制率；测定0、10．5、14．O、28．Oramol／LCENPOANU对MCF-7的克隆形成率；观察14．0gmoFLCENPOANU作用O、24、48h后MCF-7的细胞形态变化和凋亡形态，以及作用48h后空白对照组和CENPOANtJ（14．0p．mol／L）组MCF-7的细胞周期分布、增殖指数和凋亡指数（A）。结果：在MCF．7、HeLa、MEL、U251上的ICmCENPOANU分别为（6．9±2．2）、（14．7±3．5）、（7．8±1．2）、（9．6±2．1）gm01／L，BCNU分别为（14．5±2．6）、（12．4±1．5）、（8．8±1．3）、（11．2±1．4）p．mol／L；CENPOANU对MCF一7的生长具有抑制作用，且与浓度和时间呈正相关。与BCNU比较，CENPOANU对MCF．7的生长抑制作用明显增强（P〈0．01）。CENPOANU能明显降低MCF-7的克隆形成率，且与浓度呈正相关（P〈0．05）。与作用0h比较，MCF．7随作用时间延长，可见体积缩小、空泡等凋亡细胞，且细胞数量减少，发生皱缩、脱落等，其中GJM、S期细胞明显减少（P〈0．01），空白对照组和CENPOANU组MCF-7的增殖指数分剐为92．05％、42．70％，A分别为（1．5±0．1）％、（13．1±1．3）％。结论：CENPOANU主要通过阻滞细胞周期G2／M、S进程干预细胞增殖，并诱导细胞凋亡，且体外抗肿瘤活性优于BCNU。

Research on Antineoplastic Activity of Anti Tumor Compound CENPOANU in vitro

OBJECTIVE： To study the antineoplastic activity of anti tumor compound 1-（2-chloroethyl）-3-2-（2-nitrophenoxy） acetyl]-l-nitrosourea （CENPOANU） in vitro and its mechanism. METHODS： The Half inhibitory concentration （1C~0） values of CENPOANU and carmustine （BCNU） were evaluated in MCF-7, HeLa, MEL and U251 cell lines. The growth inhibitory actions of CENPOANU （0, 0.5, 1.0, 1.5, 2.0 and 4.0 folds of IC~0） on MCF-7 were determined after 24 h, 36 h and 48 h. The growth in- hibitory actions of CENPOANU and BCNU （0, 0.5, 1.0, 1.5, 2.0 and 4.0 folds of IC50on MCF-7 were observed after 48 h. The effects of 0, 10.5, 14.0 and 28.0 μmol/L CENPOANU on MCF-7 clone formation rate were tested. The morphological changes and apoptosis of MCF-7 were observed in normal control group and trial group （14.0 μmol/L CENPOANU） after 0, 24, 48 h. The dis- tribution of cell cycle, proliferation index and apoptosis index （A） was observed in control group and trial group after 48 h. RE- SULTS： In MCF-7, HeLa, MEL and U251, IC50 values of CENPOANU were （6.9 ± 2.2）, （14.7 ± 3.5）, （7.8 ± 1.2） and （9.6 ± 2.1） μmol/L; IC50 values of BCNU were （14.5 ±2.6）, （12.4± 1.5）, （8.8 ± 1.3） and （11.2 ± 1.4） μmol/L. CENPOANU showed in- hibitory action on the growth of MCF-7, in concentration and time-dependant manner. Compared with BCNU, CENPOANU had strong inhibitory effect on the growth of MCF-7 （P〈0.01）. CENPOANU could inhibit the MCF clone formation rate significantly, in concentration-dependant manner （P〈0.05）. Compared with effects of 0 h, apoptotic MCF-7 cells as volume reduction and vacu- ole could be found in trial group as time went on; the reduction of cells, shrinkage and detachment were also found, and the cells of GdM, S phase decreased （P〈0.01）. Proliferation index of MCF-7 in blank control group and CENPOANU group were 92.05 % and 42.70%, and A were （1.5 ± 0.1）% and（13.1± 1.3）%, respectively. CONCLUSIONS： CENPOANU has better antineoplastic activity than BCNU in vitro though blocking the GJM, S phase of cell cycle, intervening in cell proliferation and inducing apoptosis.