Department of Pharmacology, Toxicology and Therapeutics, and Medicine, The University of Kansas Cancer Center, The University of Kansas Medical Center, Kansas City, Kansas, United States of America.
Department of Biochemistry, University of Texas Health Science Center at Tyler, Tyler, Texas, United States of America.
PLoS One. 2014 Apr 2;9(4):e92161. doi: 10.1371/journal.pone.0092161. eCollection 2014.
Pancreatic cancer is a deadly disease, and therefore effective treatment and/or prevention strategies are urgently needed. The objectives of this study were to examine the molecular mechanisms by which embelin inhibited human pancreatic cancer cell growth in vitro, and xenografts in Balb C nude mice, and pancreatic cancer cell growth isolated from KrasG12D transgenic mice. XTT assays were performed to measure cell viability. AsPC-1 cells were injected subcutaneously into Balb c nude mice and treated with embelin. Cell proliferation and apoptosis were measured by Ki67 and TUNEL staining, respectively. The expression of Akt, and Sonic Hedgehog (Shh) and their target gene products were measured by the immunohistochemistry, and Western blot analysis. The effects of embelin on pancreatic cancer cells isolated from 10-months old KrasG12D mice were also examined. Embelin inhibited cell viability in pancreatic cancer AsPC-1, PANC-1, MIA PaCa-2 and Hs 766T cell lines, and these inhibitory effects were blocked either by constitutively active Akt or Shh protein. Embelin-treated mice showed significant inhibition in tumor growth which was associated with reduced expression of markers of cell proliferation (Ki67, PCNA and Bcl-2) and cell cycle (cyclin D1, CDK2, and CDK6), and induction of apoptosis (activation of caspase-3 and cleavage of PARP, and increased expression of Bax). In addition, embelin inhibited the expression of markers of angiogenesis (COX-2, VEGF, VEGFR, and IL-8), and metastasis (MMP-2 and MMP-9) in tumor tissues. Antitumor activity of embelin was associated with inhibition of Akt and Shh pathways in xenografts, and pancreatic cancer cells isolated from KrasG12D mice. Furthermore, embelin also inhibited epithelial-to-mesenchymal transition (EMT) by up-regulating E-cadherin and inhibiting the expression of Snail, Slug, and ZEB1. These data suggest that embelin can inhibit pancreatic cancer growth, angiogenesis and metastasis by suppressing Akt and Shh pathways, and can be developed for the treatment and/or prevention of pancreatic cancer.
胰腺癌是一种致命的疾病，因此迫切需要有效的治疗和/或预防策略。本研究的目的是研究 Embelin 在体外抑制人胰腺癌细胞生长、Balb C 裸鼠异种移植瘤以及 KrasG12D 转基因小鼠胰腺癌细胞生长的分子机制。通过 XTT 测定法测量细胞活力。将 AsPC-1 细胞皮下注射到 Balb c 裸鼠中，并给予 Embelin 治疗。通过 Ki67 和 TUNEL 染色分别测量细胞增殖和细胞凋亡。通过免疫组织化学和 Western blot 分析测量 Akt 和 Sonic Hedgehog（Shh）及其靶基因产物的表达。还检查了 Embelin 对来自 10 个月大的 KrasG12D 小鼠的胰腺癌细胞的影响。Embelin 抑制胰腺癌细胞系 AsPC-1、PANC-1、MIA PaCa-2 和 Hs 766T 的细胞活力，这些抑制作用可被组成型激活 Akt 或 Shh 蛋白阻断。Embelin 治疗的小鼠肿瘤生长显著受到抑制，这与细胞增殖标志物（Ki67、PCNA 和 Bcl-2）和细胞周期标志物（cyclin D1、CDK2 和 CDK6）的表达减少以及细胞凋亡诱导（caspase-3 激活和 PARP 切割，Bax 表达增加）有关。此外，Embelin 抑制肿瘤组织中血管生成标志物（COX-2、VEGF、VEGFR 和 IL-8）和转移标志物（MMP-2 和 MMP-9）的表达。Embelin 的抗肿瘤活性与异种移植瘤和来自 KrasG12D 小鼠的胰腺癌细胞中 Akt 和 Shh 通路的抑制有关。此外，Embelin 还通过上调 E-钙粘蛋白和抑制 Snail、Slug 和 ZEB1 的表达来抑制上皮间质转化（EMT）。这些数据表明，Embelin 通过抑制 Akt 和 Shh 通路可以抑制胰腺癌细胞生长、血管生成和转移，可用于治疗和/或预防胰腺癌。
