中医学报2018，Vol.33Issue(4)：511-516,6.DOI:10.16368/j.issn.1674-8999.2018.04.123
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
摘要
Abstract
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 -1g·mL-1,10 -2g·mL-1,10 -3g·mL-1and 10 -4g·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 dy-namic 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 genera-tion of them were scattered.The cells were larger with low transparency and increased particles; when the concentration of Tianma granules was 10 -4g·mL-1,the SW480 cells visibly turned deeper in color,some cells dropped from the walls and there were de-structed and fragmented cells; in the treatment groups of 10 -1g·mL-1,10 -2g·mL-1,10 -3g·mL-1and 10 -4g·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 proliferation inhibiting rate increased (P ＜0.05 ); in the treatment groups of 10 -1g·mL-1,10 -2g·mL-1,10 -3g·mL-1and 10 -4 g·mL-1,the Tianma granules increased the AKP specific activity of SW480 cells,which appeared to be significantly the highest at 10 -2g·mL-1(P＜0.05 or P ＜0.01).No significant difference of AKP specific activity appeared in the positive control group from the blank control group (P＞0.05) ; Ki-67 and PCNA genes mainly expressed in the nucleus of SW480 cells,and the positive cells in the treatment group and positive control group were obviously less than the blank control group,with significantly decreased average expression area and integral optical density (P ＜0.01).Conclusion: Tianma granules can significantly inhibit the prolifer-ation of human colon cancer cell line SW480,which may be related to the induction of maturation and differentiation of SW480 cells or the inhibition of Ki-67 and PCNA gene expression in SW480 cells.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Key words
human colon cancer cell line SW480/Tianma granule/5-fluorouracil/cell proliferation/Ki-67 gene/PCNA gene/alkaline phosphatase specific activity分类
医药卫生引用本文复制引用
周晓丽,高宗跃..天马颗粒剂抑制人结肠癌细胞株SW480增殖机制研究[J].中医学报,2018,33(4):511-516,6.基金项目
2016年度河南省中医药科学研究专项课题(2016ZY2061) （2016ZY2061）
