OBJECTIVE: To determine the effects of PA and its glucopyranose ester, pomolic acid-28-O-β-D-glucopyranosyl ester (PAO), on colon cancer HT-29 cells.
METHODS: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay was used to measure cell viability. Apoptosis was detected via hoechst 33342 staining. PI single staining was identified by flow cytometry to determine the cycle and scratch assay was used to observe the migration of HT-29 cells. The levels of mRNA and proteins were evaluated by q polymerase chain reaction and western blotting, respectively.
RESULTS: PA and PAO considerably inhibited the growth of the HT-29 cell line in a time and dose-dependent manner. After the administration of PA and PAO for 24 and 48 h, cell apoptosis was significantly promoted and HT-29 cells were arrested in the G0/G1 stage. The Bax/Bcl2 ratio was also increased, which activated cysteinyl aspartate specific proteinase 3, leading to apoptosis; it also increased the expression of light chain 3 II/I and Beclin1, which activated autophagy and caused cell death. This in turn increased the expression of p62 to promote cell apoptosis, inhibiting the levels of signal transducer and activator of transcription 3 (STAT3) and p-STAT3, suppressing the level of Bcl2, and promoting cell.
CONCLUSIONS: Both PA and PAO provide novel therapeutic strategies for treating colorectal cancer.
目的:为了确定PA及其吡喃葡萄糖酯的作用,多酚酸-28-O-β-D-吡喃葡萄糖基酯(PAO),结肠癌HT-29细胞。
方法:3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物测定法用于测量细胞活力。通过hoechst33342染色检测细胞凋亡。通过流式细胞术鉴定PI单染色以确定周期,并使用划痕测定法观察HT-29细胞的迁移。通过q聚合酶链反应和蛋白质印迹法评估mRNA和蛋白质的水平,分别。
结果:PA和PAO以时间和剂量依赖性方式显著抑制HT-29细胞系的生长。PA和PAO给药24和48小时后,细胞凋亡显著促进,HT-29细胞阻滞在G0/G1期。Bax/Bcl2比值也增加,它激活半胱氨酰天冬氨酸特异性蛋白酶3,导致细胞凋亡;它还增加了轻链3II/I和Beclin1的表达,从而激活自噬并引起细胞死亡。这反过来增加p62的表达以促进细胞凋亡,抑制信号转导和转录激活因子3(STAT3)和p-STAT3的水平,抑制Bcl2的水平,促进细胞。
结论:PA和PAO都为治疗结直肠癌提供了新的治疗策略。