天马颗粒剂抑制人结肠癌细胞株SW480增殖机制研究  

Study on the Mechanism of Tianma Granule Inhibiting the Proliferation of Human Colon Cancer Cell Line SW480

作　　者：周晓丽[1] 高宗跃[2] ZHOU Xiaoli;GAO Zongyue(The Third Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou Henan China 450008;The Second Affiliated Hospital of Henan University of Chinese Medicine/Henan Province Hospital of Chinese Medicine, Zhengzhou Henan China 450002)

机构地区：[1]河南中医药大学第三附属医院,河南郑州450008 [2]河南中医药大学第二附属医院/河南省中医院,河南郑州450002

出　　处：《中医学报》2018年第4期511-516,共6页Acta Chinese Medicine

基　　金：2016年度河南省中医药科学研究专项课题(2016ZY2061)

摘　　要：目的:探讨天马颗粒剂抑制人结肠癌细胞株SW480增殖的作用,并对其增殖抑制作用机制进行初步探讨。方法:采用不同质量浓度的天马颗粒剂(10^(-1)g·mL^(-1)、10^(-2)g·mL^(-1)、10^(-3)g·mL^(-1)及10^(-4)g·mL^(-1))处理人结肠癌细胞株SW480,同时设立空白对照组(RPMI 1640培养液处理)及阳性对照组(以5-氟尿嘧啶处理)。观察SW480细胞的形态学动态变化,以MTT法检测各组SW480细胞的增殖情况,另以速率法检测各组SW480细胞中碱性磷酸酶(alkaline phosphatase,AKP)比活性的变化情况。建立人结肠癌细胞株SW480免疫抑制小鼠肾包膜下移植瘤模型,同时设立天马颗粒剂组(天马颗粒剂灌胃),空白对照组(生理盐水灌胃)及阳性对照组(5-氟尿嘧啶腹腔注射),8 d后采集肿瘤组织,以免疫组织化学法及图像分析技术定量检测各组移植瘤SW480细胞中Ki-67及PCNA基因的表达情况。结果:随着天马颗粒剂质量浓度的增加,SW480细胞逐渐稀疏,散在生长,细胞变大,透明度差,颗粒增多;当天马颗粒剂质量浓度为10^(-4)g·mL^(-1)时,SW480细胞颜色加深,且部分细胞从瓶壁脱落,甚至部分细胞可见坏死崩解的碎片;实验组10^(-1)g·mL^(-1)、10^(-2)g·mL^(-1)、10^(-3)g·mL^(-1)及10^(-4)g·mL^(-1)的天马颗粒剂均可显著抑制SW480细胞增殖,且随天马颗粒剂作用时间延长及质量浓度增大,SW480活细胞计数逐渐降低(P<0.05);随着质量浓度的增加OD值逐渐降低,增殖抑制率逐渐升高(P<0.05);10^(-1)g·mL^(-1)、10^(-2)g·mL^(-1)、10^(-3)g·mL^(-1)及10^(-4)g·mL^(-1)的天马颗粒剂均会使SW480细胞的AKP比活性升高,且以10^(-2)g·mL^(-1)时的AKP比活性最高,且显著高于其他各组(P<0.05或P<0.01)。阳性对照组AKP比活性与空白对照组比较,差异无统计学意义(P>0.05);Ki-67及PCNA基因主要表达于SW480细胞的细胞核,天马颗粒剂组及阳性对照组Ki-67及PCNA基因阳性着色较空白对照组明显减少,其表达的面积均�Objective： To investigate the inhibitory effect of Tianma granule on the proliferation of human colon cancer cell line SW480 and its mechanism. Methods： The human colon cancer cell line SW480 was treated with different concentrations of Tianma granules（ 10-1 g· mL-1,10-2g · mL-1,10-3 g · mL-1 and 10-4 g · mL-1） as the treatment group,the blank control group（ RPMI 1640 nutrient solution treatment） and positive control group（ 5-fluorouracil treatment） were established as well. The dynamic morphology changes of SW480 cells were observed,and the proliferation of SW480 cells of each group was tested by means of MTT and the changes of alkaline phosphatase（ AKP） specific activity were tested by velocity. A model was established for the transplanted tumor of human colon cancer cell line SW480 by subrenal capsule assay（ SRCA）,and meanwhile the treatment group（ Tianma granules gavage）,blank control group（ normal slaine gavage） and positive control group（ 5-fluorouracil intraperitoneal injection） were established. Then the tumor tissues were collected 8 days later and the Ki-76 and PCNA genetic expressions in the SW480 cells of the transplanted tumor of each group were checked with immunohistochemistry and image analysis technique by quantitative determination. Results： With the increase of Tiamna granule concentration,the SW480 cells got sparse and the generation of them were scattered. The cells were larger with low transparency and increased particles; when the concentration of Tianma granules was 10-4 g·mL-1,the SW480 cells visibly turned deeper in color,some cells dropped from the walls and there were destructed and fragmented cells; in the treatment groups of 10-1 g·mL-1,10-2 g·mL-1,10-3 g·mL-1 and 10-4 g·mL-1,the Tianma granules could significantly inhibit the proliferation of SW480 cells,and the cells alive got less with time and the increased concentration of Tianma granule concentration（ P〈 0. 05）; with the increased concentration,OD decreased,and the proliferat

关 键 词：人结肠癌细胞株SW480 天马颗粒剂 5-氟尿嘧啶 细胞增殖 KI-67基因 PCNA基因 碱性磷酸酶比活性 

分 类 号：R273.53[医药卫生—中西医结合]