背景:二十多年来,维生素D是否在癌症频率中起作用的问题,发展,死亡是有预谋的。结肠直肠,乳房,前列腺癌一直是一个谨慎的中心,总之,在美国,这三种恶性肿瘤报告了大约35%的癌症病例和20%的癌症死亡,因此,这是一种主要的公共卫生担忧。目的是以综合方法评估维生素D纳米乳(NVD)在结直肠癌细胞系和HCT116异种移植模型中的抗肿瘤活性。
方法:两种人类结直肠癌细胞系HCT116和HT29(从药学院获得,沙特国王大学,KSA长大了。进行3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物方案以显示NVD和β-连环蛋白抑制剂(FH535)对HCT116和HT29细胞系的活力的影响。进行细胞凋亡/细胞周期测定。用FACScan(Becton-Dickinson,NJ).每个样品收获约10,000个细胞,并用ModiFitLT软件(veritySoftwareHouse,我,美国)。蛋白质印迹和RT-PCR分别在体外和体内进行蛋白质和基因表达。
结果:我们发现NVD以剂量依赖的方式和时间依赖的方式诱导结直肠细胞的细胞毒性。Further,我们的数据证实,NVD给予人类结直肠癌HCT116和HT29细胞导致细胞生长停滞,调节细胞周期的分子在细胞周期的G2期起作用,并以剂量依赖的方式改变凋亡。进一步我们的结果得出结论,NVD给药降低β-catenin基因的表达,体内外AKT基因和Survivin基因及卵白表达。
结论:我们的研究结果表明,靶向β-catenin基因可能会促进细胞周期和细胞周期调节因子的改变。Wnt/β-catenin信号通路可能通过调控细胞周期和细胞周期调控因子的表达参与结直肠癌细胞的发生发展。
BACKGROUND: More than the two decades, the question of whether vitamin D has a role in cancer frequency, development, and death has been premeditated in detail. Colorectal, breast, and prostate cancers have been a scrupulous spot of center, altogether, these three malignancies report for approximately 35% of cancer cases and 20% of cancer demises in the United States, and as such are a chief public health apprehension. The aim was to evaluate antitumor activity of Vitamin D-Nanoemulsion (
NVD) in colorectal cancer cell lines and HCT116 xenograft model in a comprehensive approach.
METHODS: Two human colorectal cancer cell lines HCT116 and HT29 (gained from College of Pharmacy, King Saud University, KSA were grown. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazoliumbromide protocol were performed to show the impact of
NVD and β-catenin inhibitor (FH535) on the viability of HCT116 and HT29 cell lines. Apoptosis/cell cycle assay was performed. Analysis was done with a FACScan (Becton-Dickinson, NJ). About 10,000 cells per sample were harvested and Histograms of DNA were analyzed with ModiFitLT software (verity Software House, ME, USA). Western blotting and RT-PCR were performed for protein and gene expression respectively in in vitro and in vivo.
RESULTS: We found that
NVD induced cytotoxicity in colorectal cells in a dose-dependent manner and time dependent approach. Further, our data validated that
NVD administration of human colorectal cancer HCT116 and HT29 cells resulted in cell growth arrest, alteration in molecules regulating cell cycle operative in the G2 phase of the cell cycle and apoptosis in a dose dependent approach. Further our results concluded that
NVD administration decreases expression of β-catenin gene, AKT gene and Survivin gene and protein expression in in vitro and in vivo.
CONCLUSIONS: Our findings suggest that targeting β-catenin gene may encourage the alterations of cell cycle and cell cycle regulators. Wnt/β-catenin signaling pathway possibly takes part in the genesis and progression of colorectal cancer cells through regulating cell cycle and the expression of cell cycle regulators.