摘要
目的探讨姜黄素体内对人结肠癌耐长春新碱细胞株HCT-8/VCR多药耐药的逆转作用及其可能机制。方法将HCT-8/VCR种植于裸鼠皮下,建立肿瘤耐药模型。移植瘤裸鼠20只依据治疗方法的不同分为对照组(PBS)、长春新碱(Vincristine,VCR;2 mg/kg)组、姜黄素(50 mg/kg)组、姜黄素联合VCR(50 mg/kg姜黄素+2 mg/kg VCR)4组,每组5只。治疗2周后收集标本,称量各组瘤体质量,采用RT-PCR、Western blot分别检测瘤组织内MDR1 mRNA、survivin mRNA和P-gp、survivin蛋白的表达水平,HE染色观察瘤组织细胞形态学变化,评估姜黄素在不同处理因素中逆转肿瘤多药耐药的效果。结果姜黄素联合VCR能显著抑制瘤体的生长,姜黄素联合VCR组瘤质量[(0.42±0.23)g]明显低于对照组[(1.21±0.25)g]、VCR组[(1.13±0.27)g]、姜黄素组[(0.75±0.27)g](P<0.05)。RT-PCR、Western blot结果显示姜黄素能够下调MDR1 mRNA、survivin mRNA和P-gp、survivin蛋白的表达,姜黄素联合VCR组MDR1 mRNA(0.18±0.05)、survivin mRNA(0.63±0.04)、P-gp(0.15±0.02)、survivin(0.31±0.09)和姜黄素组MDR1 mRNA(0.39±0.11)、survivinmRNA(0.80±0.12)和P-gp(0.34±0.07)、survivin(0.40±0.11)蛋白表达明显低于对照组MDR1 mRNA(0.99±0.23)和survivin mRNA(1.19±0.32)表达,以及P-gp(0.71±0.23)、survivin(0.88±0.15)蛋白表达(P<0.05)。HE病理切片显示姜黄素组和姜黄素联合VCR组较其他2组有较多的淋巴细胞浸润,核固缩较多,核分裂相少,坏死更为广泛。结论多种机制参与姜黄素体内逆转肿瘤多药耐药,姜黄素可能通过下调MDR1、survivin mRNA表达和P-gp、survivin蛋白的表达,促进肿瘤细胞凋亡等多种途径逆转肿瘤的多药耐药性。
Objective To investigate the multi-drug resistance reversal effects of curcumin on xenografts derived from multi-drug resistant human colon carcinoma cell line HCT-8/VCR,and to explore the possible mechanism.Methods The models of HCT-8/VCR xenograft（100 mm3） in nude mice were established through implanting 0.1 ml cells at 4×107 cells/ml.Twenty nude mice bearing HCT-8/VCR tumors were randomized into 4 groups according to different treatments,that is,group A： control group,group B： VCR group（2 mg/kg injected around the tumor mass,once per 2 d,for 7 times）,group C： curcumin group（50 mg/kg,injected around the tumor mass,once per 2 d,for 7 times）,group D： curcumin plus VCR group（as combined the treatment of groups B and C）.Tumor volume was calculated with L×D2/2 which were measured once per 2 d,for 14 d.Tumor weight was measured and specimens were collected from the mice in each group in 2 weeks after the interventions.Morphology of xenografts were investigated by HE staining.The expressions of MDRl,P-glycoprotein（P-gp） and survivin were observed with reverse transcriptase PCR and Western blot assay respectively.Effects of curcumin on reversing multi-drug resistance of tumor in different treatments were evaluated.Results Curcumin plus VCR significantly inhibited the growth of xenografts,with the tumor weight of group D（0.42±0.23 g） significantly lower than those of the other 3 groups（group A 1.21±0.25 g,group B 1.13±0.27 g and group C 0.75±0.27 g,P0.05）.RT-PCR and Western blotting results showed that the expressions of MDRl mRNA（0.39±0.11）,P-gp（0.34±0.07）,survivin mRNA（0.80±0.12） and protein（0.40±0.11） in group C and those of MDRl mRNA（0.18±0.05）,P-gp（0.15±0.02）,survivin mRNA（0.63±0.04） and protein（0.40±0.11） in group D were significantly lower than those in group A（MDRl mRNA 0.99±0.23,P-gp 0.71±0.23,survivin mRNA 1.19±0.32 and protein 0.88±0.15,P0.05）,indicating that curcumin down regulated these expressions.HE staining revealed that group C and D had more lymphocytic infiltration,more nuclear pyknosis,fewer mitotic figures,and more extensive necrosis when compared to the other groups.Conclusion Curcumin has strong reversal effects on multidrug resistance in HCT-8/VCR cells through down-regulating mRNA and protein expressions of MDR1 and survivin.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2011年第4期376-380,共5页
Journal of Third Military Medical University
基金
重庆市卫生局中医药重点项目(2009-1-2)~~
关键词
姜黄素
抗药性
多药
移植
异种
肿瘤
逆转
小鼠
裸
curcumin
multidrug resistance
xenograft
neoplasms
reversal
nude mice
