大黄素通过NF-κB信号途径抑制胰腺癌生长和转移

Anti-Proliferation and Anti-Metastasis Effects of Emodin on Human Pancreatic Cancer via NF-( B Signaling Pathway

目的：观察大黄素抑制体内外胰腺癌生长和转移的作用及其机制。方法：以不同浓度的大黄素（10～40μmol·L^-1）作用胰腺癌BxPC-3细胞24h后，应用CellCountingKit-8（CCK-8）法检测细胞增殖；流式细胞术（FCM）及膜联蛋白V（AnnexinV）／碘化丙啶（PI）双染色法检测大黄素作用后细胞凋亡率的变化；划痕试验和Matrigel细胞侵袭实验分别测定胰腺癌细胞体外迁移和侵袭能力；凝胶电泳迁移率实验（EMSA）检测胰腺癌细胞中NF-κB活性表达水平的变化；Westernblot法检测药物作用后细胞中蛋白表达；裸鼠胰腺癌原位移植模型建立后2周，使用大黄素进行干预治疗，采用免疫组化方法检测Ki-67、NF-κB、Survivin和MMP-9的表达。结果：与对照组相比较，大黄素可呈浓度依赖性抑制BxPC-3细胞生长、迁移和侵袭；大黄素可上调BxPC-3细胞中Caspase-3表达，并下调Survivin和MMP-9的表达水平，呈现出明显的量效关系；体内试验结果表明，大黄素可抑制体内胰腺癌生长和转移，并降低肿瘤组织中Ki-67、NF-κB、Survivin和MMP-9的表达。结论：大黄素可显著抑制体内外胰腺癌生长和转移，通过抑制NF-κB及其调控蛋白Survivin和MMP-9可能是其重要作用机制之一。

Objective：To investigate the anti - proliferation and anti - metastasis effects of emodin on the pancreatic cancer in vitro and in vivo. Methods ： Human pancreatic cancer cell line BxPC - 3 was treated with different concentrations of emodin and the cellular proliferation was detected by Cell Counting Kit - 8 （ CCK - 8 ） assay. The flow cytometry （FCM） was used to determine apoptosis in pancreatic cancer ceils. The effects of emodin on the migration and invasion of BxPC - 3 cells were examined by using wound assay and matrigel counting. The activity of NF - κB in pancreatic cancer cells was measured by electrophoretic mobility shift assay （EMSA）. Western blot was used to detect the protein expression in pancreatic cancer cells. Metastatic model simulating human pancreatic cancer was established by orthotropic implanta- tion of histologically intact human tumor tissue into pancreatic wall of nude mice. Immunohistochemistry was used to detect the protein expression in the tumors. Results ： Emodin induced a higher percentage of growth inhibition and apoptosis in pancreatic cancer cell line BxPC - 3 than that of control. Emodin suppressed the migration and invasion of BxPC - 3 ceils in a dose - dependent manner. Emodin can up - regulate NF -κB DNA - binding activity and down - regulate survivin and MMP - 9 in BxPC - 3 cells. The expression of Caspase - 3 was up - regulated in BxPC - 3 cells after treatment of emodin. Emodin showed significant decrease in tumor weight and metastasis compared to untreated control. Conclusion： Emodin exerts anti - proliferative and anti - metastatic activity in pancreatic cancer both in vitro and in vivo, which may be related to down - regulation of NF - κB and its regulated molecules such as survivin and MMP - 9 proteins.