膈下逐瘀汤逆转裸鼠移植瘤MDR-1表达的实验

Experiment of Diaphragmatic Stasis Expelling Decoction Reversing MDR-1 Expression in Transplantation Tumor in Nude Mice

目的探讨膈下逐瘀汤对HCT-8/5-Fu裸鼠移植瘤多药耐药基因MDR-1表达的影响。方法 MTT法确认人结肠腺癌细胞HCT-8/5-Fu的耐药性及对其他化疗药VCR、VP-16、DDP的交叉耐药性后,将耐药细胞HCT-8/5-Fu接种于BABL/c无胸腺裸鼠右腋下,建立耐药性稳定的裸鼠移植瘤模型。移植瘤裸鼠模型随机分为四组:空白对照组(对照组)、5-Fu组、膈下逐瘀汤组(DSED组)、膈下逐瘀汤+5-Fu组(DSED+5-Fu组)。其中对照组予0.9%氯化钠溶液灌胃给药,其余组予腹腔注射5-Fu和(或)中药灌胃。连续给药14天后颈椎脱臼处死裸鼠。观察各组裸鼠移植瘤生长情况;RT-PCR法检测各瘤组织MDR-1 mRNA表达。结果膈下逐瘀汤+5-Fu组、膈下逐瘀汤组、5-Fu组对裸鼠移植瘤的抑瘤率分别为54.9%、32.4%和7.0%(P<0.01)。RT-PCR结果显示膈下逐瘀汤+5-Fu组及膈下逐瘀汤组MDR-1 mRNA的表达量与对照组相比明显下降(P<0.01),5-Fu组与对照组相比MDR-1 mRNA表达量略有升高,但差异无统计学意义。结论膈下逐瘀汤对大肠癌多药耐药裸鼠移植瘤生长具有抑制作用,其机制可能与膈下逐瘀汤通过逆转耐药移植瘤多药耐药基因表达,增强肿瘤细胞对药物的敏感度。

Objective To investigate the effects of diaphragmatic stasis expelling decoction on the multidrug resistance gene expression in transplantable tumor cells in nude mice bearing cell line HCT-8/5-Fu. Methods MTT was applied to confirm the drug resistance of human colonic carcinoma cell line HCT-8/5-Fu and the cross resistance towards other chemotherapy drugs, such as VCR, VP-16 and DDP. The model of transplantable tumor in nude mice with stable drug resistance was established by planting resistant cell line HCT-8/5-Fu into the right armpit of BABL/c nude mice without thymus. A total of 28 nude mice were randomly divided into four groups: control group, diaphragmatic stasis expelling decoction group(DSED group), 5-Fu group and diaphragmatic stasis expelling decoction plus 5-Fu group(DSED+5-Fu group). The control group received an intragastric administration of 0.9% sodium chloride solution and others received an intraperitoneal injection of 5-Fu or an intragastric administration of traditional Chinese medicine. Drug delivery was lasted for 14 days before the mice were executed. The growth of transplantable tumors in nude mice in all groups was observed. MDR-1 expression in all tumor tissues was detected by RT-PCR. ResultsTumor inhibition rates of DSED+5-Fu group, DSED group and 5-Fu group were 54.9%, 32.4%, and 7.0% respectively(P<0.01). RT-PCR results showed that the expression amounts of MDR-1 mRNA in DSED+5-Fu group and DSED group were obviously less than that in control group(P<0.01). The expression amount of MDR-1 mRNA in 5-Fu group was increased slightly compared with the control group with no statistical significance. Conclusion Diaphragmatic stasis expelling decoction has inhibitory effect on the growth of transplantable tumor in nude mice with multi-drug resistance to colon carcinoma. The mechanism may be related to diaphragmatic stasis expelling decoction reversing MDR-1 mRNA expression of transplantable tumor in nude mice, therefore to enhance the sensitivity to drugs of tumor cells.