Hu Gaoyu, Huang Zansong, Zhou Xihan, Hu Jing, Huang Bingchen
Institute of Digestive Disease, Department of Gastroenterology, the Affiliated Hospital of Youjiang Medical College for Nationalities, Baise 533000, China.
Zhonghua Gan Zang Bing Za Zhi. 2015 Sep;23(9):669-74. doi: 10.3760/cma.j.issn.1007-3418.2015.09.007.
To investigate the effect and molecular mechanism of cisplatin (DDP) combined with Matrine (Ma;plant alkaloid) against hepatocellular carcinoma using a nude mouse model with xenografted human tumors.
Twenty-four 6-week old male BALB/c nude mice were subcutaneously injected with HepG2 cells into the axilla, and randomly divided into four groups:control (NS) group,Ma treatment group,DDP treatment group and DDP+Ma combination treatment group. All treatments were delivered via intraperitoneal injection.Changes in whole body weights and tumor volume were assessed by before and after treatment measurements and plotting of growth curves. After 14 days of drug intervention, the mice were sacrificed for collection of tumor tissue and assessment of the tumor inhibition rates for each treatment. Affects on expression of survivin and caspase-3 were assessed by immunohistochemistry. ANOVA test and t-test were performed for the statistical analyses.
The tumor inhibition rates for the various treatments were:37.5%,Ma alone;75.0% DDP alone;83.3%,DDP+Ma group DDP combined. The DDP+Ma-induced inhibition was significantly greater than that achieved wit Ma or DDP alone (both P less than 0.05). The average weight of the DDP+Ma group (21.5 g) was lower than that of the NS group (28.5 g) and the Ma group (26.67 g),but higher than that of the DDP group (17.33 g).In addition, the DDP+Ma group also showed more robust general health,as indicated by activity,participation in life routines and appetite,than the DDP group. The rate of positive staining for survivin expression in tumor tissues was significantly lower in the DDP+Ma group (19.58%+/-4.52%) than in the NS group (83.26%+/-15.56%), the Ma group (62.50%+/-8.09%), and the DDP group (38.67%+/-8.26%) (all P less than 0.05).In contrast, the rate of positive staining for Bax expression was significantly higher in the DDP+Ma group (78.26%+/-6.09%) than in the NS group (21.15%+/-3.68%), the Ma group (35.13%+/-10.57%), and the DDP group (65.88%+/-4.81%) (all P less than 0.05).
Treatment with Ma alone or DDP alone is sufficient to inhibit the growth ofxenografted human hepatocellular carcinoma cells in nude mice. The DDP+Ma combination treatment,however,shows greater inhibitory effect,suggesting that Ma may enhance DDP's anticancer properties. The improved health status of mice treated with DDP+Ma suggests that Ma may reduce DDP toxicity. The mechanism underlying these beneficial treatment effects may involve modulation of survivin/caspase-3 expression and subsequent apoptosis.
利用人肿瘤异种移植裸鼠模型，研究顺铂（DDP）联合苦参碱（Ma；植物生物碱）对肝癌的作用及其分子机制。
将24只6周龄雄性BALB/c裸鼠腋窝皮下注射HepG2细胞，随机分为四组：对照组（NS）、Ma治疗组、DDP治疗组和DDP+Ma联合治疗组。所有治疗均通过腹腔注射给药。通过治疗前后测量体重和绘制生长曲线来评估全身重量和肿瘤体积的变化。药物干预14天后，处死小鼠以收集肿瘤组织并评估每种治疗的肿瘤抑制率。通过免疫组织化学评估对survivin和caspase-3表达的影响。进行方差分析和t检验进行统计学分析。
各种治疗的肿瘤抑制率分别为：Ma单独治疗为37.5%；DDP单独治疗为75.0%；DDP+Ma联合治疗组为83.3%。DDP+Ma联合诱导的抑制作用明显大于单独使用Ma或DDP的抑制作用（均P<0.05）。DDP+Ma组的平均体重（21.5 g）低于NS组（28.5 g）和Ma组（26.67 g），但高于DDP组（17.33 g）。此外，DDP+Ma组在活动、参与日常生活和食欲方面显示出比DDP组更强的总体健康状况。肿瘤组织中survivin表达的阳性染色率在DDP+Ma组（19.58%±4.52%）明显低于NS组（83.26%±15.56%）、Ma组（62.50%±8.09%）和DDP组（38.67%±8.26%）（均P<0.05）。相反，Bax表达的阳性染色率在DDP+Ma组（78.26%±6.09%）明显高于NS组（21.15%±3.68%）、Ma组（35.13%±10.57%）和DDP组（65.88%±4.81%）（均P<0.05）。
单独使用Ma或DDP足以抑制裸鼠体内人肝癌异种移植细胞的生长。然而，DDP+Ma联合治疗显示出更大的抑制作用，表明Ma可能增强DDP的抗癌特性。DDP+Ma治疗的小鼠健康状况改善表明Ma可能降低DDP的毒性。这些有益治疗效果的潜在机制可能涉及survivin/caspase-3表达的调节及随后的细胞凋亡。
