沙棘总黄酮抑制膀胱癌细胞系T24增殖

Total flavonoids of Hippophae rhamnoides L.inhibits proliferation of bladder cancer cell line T24

目的探讨沙棘总黄酮(HTF)对膀胱癌细胞系T24增殖和凋亡的影响及可能机制。方法体外培养T24细胞,用不同浓度(25、50和100μg/mL)HTF干预24 h后,CCK-8法、流式细胞测量术和Western blot依次检测T24细胞增殖、凋亡和蛋白Bcl-2、Bax表达,RT-qPCR检测miR-144表达。转染miR-144抑制剂至T24细胞,构建敲减miR-144表达的T24细胞,并用100μg/mL HTF干预敲减miR-144的T24细胞。上述相同方法检测敲减miR-144对T24细胞增殖和凋亡的影响。结果与对照组比较,不同浓度HTF降低了T24细胞的A值和Bcl-2表达,提高了凋亡率和Bax表达,同时促进了miR-144表达(P<0.05)。敲减miR-144的T24细胞A值和Bcl-2表达均高于未敲减miR-144的T24细胞(P<0.05),而凋亡率和Bax表达均低于未敲减miR-144的T24细胞(P<0.05)。100μg/mL HTF作用于敲减miR-144的T24细胞的A值及Bcl-2表达均高于未敲减miR-144的T24细胞(P<0.05),而凋亡率和Bax表达均低(P<0.05)于未敲减miR-144的T24细胞。结论HTF可能通过上调miR-144阻滞膀胱癌细胞增殖,并加剧了细胞凋亡。

Objective To investigate the effect of total flavonoids of Hippophae rhamnoides L.(HTF)on the proliferation and apoptosis of bladder cancer cell line T24 and underlying mechanism.Methods The bladder cancer T24 cells were incubated with different concentrations(25,50,100μg/mL)of HTF for 24 h,then CCK-8 assay,flow cytometry and Western blot were used to detect proliferation,apoptosis and the expression of Bcl-2 and Bax proteins,respectively.The expression of miR-144 was detected by RT-qPCR.miR-144 inhibitors were transfected into T24 cells to construct T24 cells whose expression of miR-144 was knocked down,then 100μg/mL HTF was used to interfere with the miR-144-knoched down T24 cells.The same methods were used to detect the effects of knocking down of miR-144 on cell proliferation and apoptosis of T24 cell.Results Compared with the control group,different concentrations of HTF reduced the cell A value and the expression of Bcl-2,and increased the apoptosis rate and the expression of Bax.At the same time,the expression of miR-144(P<0.05)was increased.The A value of T24 cells that knocked down miR-144 and the expression of Bcl-2 protein in the cells was higher than those of control T24 cells(P<0.05),but the apoptosis rate and the expression of Bax protein were lower(P<0.05).After T24 cells that knocked down miR-144 were treated with 100μg/mL HTF,their A value and the expression of Bcl-2 protein in the cells were higher than those of T24 cells that not knocked down miR-144(P<0.05),but the apoptosis rate and the expression of Bax protein were lower than those of control T24 cells(P<0.05).Conclusions HTF may slow-down the proliferation of bladder cancer cells and speed-up their apoptosis by up-regulating miR-144.