禹白附提取物抑制人胃癌细胞增殖转移的实验研究

Typhonium giganteum extract inhibits human gastric carcinoma cell proliferation and metastasis

摘要:

目的：观察禹白附提取物对人胃癌细胞GC9811细胞增殖、迁移的影响，探讨其可能的作用机制。方法：采用植物化学技术对禹白附进行提取分离，再采用四甲基偶氮唑蓝比色法（tetramethyl azo thiazole blue colorimetric method，MTT）检测不同浓度禹白附提取物作用不同时间对GC9811细胞增殖的抑制作用；细胞划痕实验和Transwell小室法检测禹白附提取物对细胞迁移能力的影响；实时荧光定量PCR和蛋白质印迹法（real-time quantitative polymerase chain reaction method，RT-qPCR）细胞基质金属蛋白酶-9（matrix metalloproteinase-9，MMP-9）、基质金属蛋白酶-2（matrix metalloproteinase-2，MMP-2）、E-钙黏蛋白（E-Cadherin）的表达水平。结果：MTT比色分析法检测结果表明，禹白附提取物对GC9811细胞的生长有明显抑制作用且浓度和作用时间呈正相关；细胞划痕实验和Transwell小室法结果显示，随禹白附提取物浓度的增加，GC9811细胞迁移能力下降；实时荧光定量PCR和蛋白质印迹法结果表明，与禹白附提取物15 g/L组比较，禹白附提取物30 g/L组的MMP-2、MMP-9表达值均明显降低，而E-Cadherin表达明显上调。结论：禹白附提取物能够抑制胃癌细胞的增殖和转移，其机制可能与其抑制MMP-9和MMP-2上调E-Cadherin的表达有关。因此，禹白附提取物具有很强的抗肿瘤作用，为进一步寻找高效低毒的抗肿瘤先导化合物奠定了基础，值得进行深入研究。

Abstract:

Objective：To examine the effects of Typhonium giganteum extract（TGE） on the proliferation and metastasis of human gastric carcinoma cell line GC9811 and to elucidate its possible mechanism of action. Methods：TGE was isolated and purified by phyto－chemistry，and the inhibitory effect of different concentrations of TGE on GC9811 cell proliferation at various time points was mea－sured by MTT colorimetric assay. The effect of TGE on cell migration was determine by cell scratch assay and Transwell migration assay. The mRNA and protein expression of matrix metalloproteinase-9（MMP-9），matrix metalloproteinase-2（MMP-2），and E-cad－herin was measured by real-time quantitative PCR and Western blot，respectively. Results：The MTT assay showed that TGE significantly inhibited the growth of GC9811 cells in a dose-dependent manner. The cell scratch and Transwell migration assays demonstrated that the migratory ability of GC9811 cells decreased as TGE concentration increased. Furthermore，MMP-2 and MMP-9 expression was significantly downregulated and E-cadherin expression was significantly upregulated in the 30 g/L TGE group than in the 15 g/L TGE group. Conclusion：TGE can significantly inhibit the proliferation and metastasis of GC9811 cells，and its mechanism of action may be related to the upregulation of E-cadherin expression and downregulation of MMP-9 and MMP-2 expression. TGE is a highly potent and minimally toxic antitumor lead compound worthy of further investigation.