健脾解毒汤对人大肠癌细胞HT-29基质裂解蛋白-9基因表达及蛋白分泌影响的实验研究  

Experimental study of Jianpi Jiedu decoction on matrixmetalloproteinase-9(MMP-9) gene expression and protein secretion in human colorectal cancer HT-29 cell

作　　者：徐磊[1] 杨静[1] 吕茜[2] 韩东[3] 

机构地区：[1]南京中医药大学附属徐州市中医院,江苏徐州221000 [2]东南大学附属徐州市中心医院中心实验室,江苏徐州221000 [3]徐州医学院麻醉学重点实验室,江苏徐州221000

出　　处：《中国肿瘤外科杂志》2014年第3期170-173,共4页Chinese Journal of Surgical Oncology

基　　金：徐州市科技发展基金计划项目(XF11C079)

摘　　要：目的探讨健脾解毒汤抗人大肠癌HT-29细胞转移的分子机制。方法健脾解毒汤加减后按功效分为基本方、清热解毒、活血化瘀、补脾益气、补血益气及扶助正气方,将各方剂水提物与人大肠癌HT-29细胞进行体外培养,四甲基氮唑蓝比色法(MTT法)检测各组细胞活力,逆转录-聚合酶链式反应(RT-PCR)和酶联免疫吸附法(ELISA)检测各组肿瘤细胞基质金属蛋白酶-9(MMP-9)mRNA基因表达和蛋白分泌的变化。结果各组中药对HT-29细胞均有不同程度的抑制作用,相同方剂组高浓度抑制作用强于低浓度,滋阴组在1 000μg/ml浓度时抑制率达到峰值(46.1%),且较基本方及清利组有显著性差异(P<0.05)。与空白对照组比较,各中药组肿瘤细胞MMP-9 mRNA基因表达水平被下调,MMP-9蛋白分泌不同程度受到抑制。其中,滋阴组抑制效果最为显著(P<0.01),较基本方及清利组亦有显著性差异(P<0.05)。结论健脾解毒汤及其加减方对大肠癌细胞的增殖、侵袭及转移具有抑制作用,通过下调肿瘤细胞MMP-9 mRNA基因表达从而抑制蛋白分泌是可能的机制之一。Objective To investigate the molecular mechanism of Jianpi Jiedu Decoction anti human colorectal cancer HT-29 cell metastasis. Methods By adding different Chinese herbal medicine, Jianpi Jiedu Decoction was divided into basic recipe and Qingre Jiedu, Huoxue Huayu, Bupi Yiqi, Buxue Yiqi, Fuzhu Zhengqi recipes according to function. Colorectal cancer HT-29 cells were cuhured with the water extract of each recipe in vitro, to detect the cell vitality by four methyl tetrazolium blue colorimetric method （MTT method） , reverse transcription polymerase chain reaction （RT-PCR） and enzyme linked immunosorbent assay （ELISA） were used to detect matrix metalloproteinase-9 （MMP-9） gene expressions and protein secretion. Results Each Chinese herbal medicine recipe had different degrees of inhibitory effect on HT-29 cells, and high concentration was stronger than low concentration in the same prescription group. Inhibition rate reached the peak at the concentration of 1 000μg/ml （46.1%） in Nourishing Yin group, and had a significant difference compared with the basic prescription group and Qing Li group （P 〈 0.05 ）. Compared with the blank control group, MMP-9 gene expression and protein secretion were down regulated in each Chinese medicine groups. The most significant effect was showed in Nourishing Yin group（P 〈 0.01 ）, and this group had significant difference compared with the basic prescription group and Qing Li group, P 〈 0.05. Conclusion Jianpi Jiedu Decoction and its related recipes have inhibitory effect on tumor cell proliferation, invasion and metastasis of colorectal cancer cell. Down regulation of MMP-9 gene expression and protein secretion may be one of the possible mechanisms.

关 键 词：健脾解毒汤 大肠癌 HT-29细胞 MMP-9 基因表达 蛋白分泌 

分 类 号：R735.34[医药卫生—肿瘤]