Colorectal cancer is the third most common malignant tumor and a leading cause of cancer death. Currently lacks effective therapies available to improve the prognosis. In the present study, VALD-3, an important Schiff base ligand from o-vanillin derivatives was evaluated for its anti-cancer activity in vitro and in vivo against colorectal cancer. The effect of VALD-3 on colorectal cancer cells proliferation was assessed using MTT assay and the cell migration was evaluated using wound healing scratch assay. The appearance of apoptotic colorectal cancer cells was detected by flowcytometry analysis. Morphological changes caused by VALD-3 induced apoptosis were also observed by Hoechst 33258 staining. The flow cytometry assay was also used to measure cell cycle arrest. The expression levels of TP53 and Bad were analyzed using quantitative real-time PCR. Protein expression of P53, Wnt/β-catenin signaling pathway proteins, apoptosis proteins and cell cycle-related protein were viewed by Western blotting. In addition, HT-29 cells xenograft tumor model was used for the study in vivo. Immunohistochemistry (IHC) staining was employed to detect the P53 protein expression. The results showed that VALD-3 obviously inhibited the proliferation and migration for colorectal cancer cells. In addition, flow cytometry analysis demonstrated that VALD-3 markedly increased early and late apoptosis on colorectal cancer cells, respectively. VALD-3 induced cell cycle arrest at the G0/G1 phase. Most importantly, tumor growth in HT-29 xenograft mice was suppressed by VALD-3, but no significant change in body weight. As confirmed by IHC staining from tumor tissue, the P53 proteins expression increased. These results suggested that VALD-3 represses cell proliferation and induces apoptosis associated with upregulating tumor suppressor activity of p53 to inhibit Wnt/β-catenin signal pathway, and it is a potential anticancer agent for colorectal cancer.

中文翻译：

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VALD-3通过上调p53抑癌活性抑制Wnt/β-catenin信号通路抑制结直肠癌细胞增殖并诱导其凋亡。

结直肠癌是第三常见的恶性肿瘤，也是癌症死亡的主要原因。目前缺乏有效的治疗方法来改善预后。在本研究中，评估了邻香兰素衍生物的重要席夫碱配体 VALD-3 在体外和体内对结直肠癌的抗癌活性。使用 MTT 法评估 VALD-3 对结直肠癌细胞增殖的影响，并使用伤口愈合划痕法评估细胞迁移。通过流式细胞术分析检测凋亡的结直肠癌细胞的出现。Hoechst 33258染色也观察到VALD-3诱导的细胞凋亡引起的形态学变化。流式细胞术测定也用于测量细胞周期停滞。使用定量实时 PCR 分析 TP53 和 Bad 的表达水平。Western blotting观察P53、Wnt/β-catenin信号通路蛋白、凋亡蛋白和细胞周期相关蛋白的蛋白表达。此外，HT-29细胞异种移植肿瘤模型用于体内研究。采用免疫组织化学 (IHC) 染色来检测 P53 蛋白的表达。结果表明，VALD-3明显抑制结直肠癌细胞的增殖和迁移。此外，流式细胞术分析表明，VALD-3分别显着增加结直肠癌细胞的早期和晚期凋亡。VALD-3 在 G0/G1 期诱导细胞周期停滞。最重要的是，HT-29 异种移植小鼠的肿瘤生长被 VALD-3 抑制，但体重没有显着变化。正如来自肿瘤组织的 IHC 染色所证实的，P53 蛋白表达增加。这些结果表明，VALD-3通过上调p53的抑癌活性来抑制Wnt/β-catenin信号通路，抑制细胞增殖并诱导细胞凋亡，是一种潜在的结直肠癌抗癌剂。