两组中药复方影响前列腺癌细胞生长的体内外实验研究

目的 探讨两组中药复方对前列腺癌细胞PC-3 体内外生长的影响及其可能的作用机制.方法 以临床常用的抗癌中药组成两组复方(复方B:白花蛇舌草、半枝莲、土茯苓和莪术;复方C:白花蛇舌草、半枝莲、土茯苓、莪术、三棱和皂角刺),分别用等量生理盐水和两组复方的水煎剂对成年雄性去势SD 大鼠灌胃后取血制成空白对照(A 组)和含药血清(B、C组);三组血清加入PC-3 的细胞培养液,应用MTT 法、流式细胞术、Hoechst 染色法检测细胞存活率、细胞周期分布及凋亡的差异.建立BALB/C 裸鼠荷前列腺癌动物模型,分别用生理盐水和两组复方的水煎剂灌胃,观察裸鼠肿瘤生长情况,HE 染色观察肿瘤组织形态学变化,TUNEL 法检测肿瘤细胞凋亡情况,免疫组化染色法检测肿瘤组织中Ki-67 表达.结果 体外细胞培养实验中B 组的细胞存活率最低,且随着培养时间的增加而持续降低;C 组的细胞存活率较B 组高,但显著低于对照组;两个复方组的S 期细胞比例显著低于对照组,而G0/G1 期细胞比例显著高于对照组(均P ＜0.05);B 组和C 组的凋亡细胞比例约为30%和10%,均多于对照组.裸鼠体内实验中B 组的抑瘤率大于C 组的抑瘤率(23% vs.18%),B 组的平均瘤重最小,平均肿瘤体积最小;B 组的TUNEL 阳性率最高,Ki-67 阳性率最低.结论 两组中药复方在体外显著抑制PC-3 细胞增殖,同时诱导其凋亡,在体内也能抑制肿瘤生长;并且复方B 的抑瘤作用最强.

Effect of two compound recipes on human hormone independent prostate cancer in vitro and in vivo

Objective To investigate the effect of two compound recipes on human hormone independent prostate cancer cell line PC-3 in vitro and in vivo,and exploring the possible mechanism.Methods The two compound recipes were composed of commonly clinically used herbal medicine（Compound recipe B：Oldenlandia diffusa,Scutellariae barbatae,Glabrous greenbrier rhizome and Zedoary rhizome.Compound recipe C：Oldenlandia diffusa,Scutellariae barbatae,Glabrous greenbrier rhizome,Zedoary rhizome,Rhizoma Sparganii and Spina Gleditsiae）.The adult male castrated SD rats in compound recipe groups were gavaged with decoction of compound recipe B or C twice a day for 5 days respectively.The animals in control group（group A）were gavaged with normal saline（NS）in the same volume as compound recipe groups.Blood samples were taken from the inferior vena cava of rats and prepared for drug-containing serum.PC-3 cells were incubated in the culture medium with the drug-containing serum and blank control serum respectively.The survival rate of the PC-3 cells was measured by MTT assay,cell cycle distribution of PC-3 was measured by flow cytometry and apoptotic rate of cells was observed under a fluorescence microscope.Prostate cancer animal models were established by subcutaneously inoculated with prostate cancer tissue in BALB/c nude mice.These nude mice were randomly divided into three groups by different gavaging methods：control group（Group A）,compound recipe B（Group B）and compound recipe C（Group C）.Conditions of tumors were evaluated by gross appearance,HE staining,TUNEL assay and immunohistochemical staining of Ki-67.Results The survival rate of Group B was the lowest and had been decreasing with the incubated time.The survival rate of Group C was higher than that of Group B,but was significantly lower than that of control group.The percentage of S-phase cells in Group B and Group C was significantly lower than that in the control group,and the percentage of G0/G1-phase cells was significantly higher than that in the control group.The percentage of apoptotic cells in Group B and Group C（30% and 10%,respectively）was significantly more than in the control group.The tumor growth suppression rates of Group B was higher than that of Group C（23% vs.18%）and the mean tumors weight and tumors volume of group B were also the lowest in the three groups.Cellular apoptosis was the most commonly happened in Group B.The expression of Ki-67 was the weakest in Group B.Conclusions The two compound recipes significantly inhibit proliferation of PC-3 cells and induce apoptosis in vitro.And both of the two compound recipes inhibit the growth of PC-3 cells in vivo.The effect of compound recipe B for controlling tumor growth was more than that of compound recipe C.