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目的研究猪苓多糖(polyporus polysaccharide,PPS)对膀胱癌T24细胞凋亡的作用及其机制。方法 T24细胞常规培养后加入不同剂量PPS,利用Hoechst染色、流式细胞术检测PPS对细胞凋亡的影响,利用q RT-PCR检测PPS对T24细胞bcl-2 m RNA表达水平及其稳定性的影响,通过Western Blot法检测PPS对T24细胞bcl-2蛋白、人抗原R(human antigen R,Hu R)蛋白的表达及对Hu R蛋白胞浆及胞核内分布的影响。结果与对照组比较,中、高剂量的PPS作用24 h后,T24细胞凋亡明显增多(P<0.05),bcl-2 m RNA稳定性降低(P<0.05),bcl-2蛋白及m RNA表达减少(P<0.05),总Hu R蛋白没有明显变化,但胞浆Hu R蛋白表达下降(P<0.05),胞核Hu R蛋白表达升高(P<0.05)。结论 PPS可诱导T24细胞凋亡,其作用可能与影响Hu R在细胞内定位,降低bcl-2 m RNA稳定性及减少bcl-2蛋白表达有关。
Objective To study the effect of polyporus polysaccharide (PPS) on apoptosis of bladder cancer T24 cells and its mechanism. Methods T24 cells were cultured routinely with different doses of PPS, and the effect of PPS on apoptosis was detected by Hoechst staining and flow cytometry. The expression of bcl-2 mRNA and its stability in T24 cells were detected by q RT-PCR The effect of PPS on the expression of bcl-2 protein, human antigen R (Hu R) protein and the distribution of Hu R protein in the cytoplasm and nucleus of T24 cells was detected by Western Blot. Results Compared with the control group, the apoptosis of T24 cells was significantly increased (P <0.05) and the stability of bcl-2 mRNA was decreased (P <0.05) after treated with medium and high doses of PPS for 24 h. The expressions of bcl-2 protein and m RNA (P <0.05). There was no significant change in total Hu R protein, but the expression of Hu R protein in cytoplasm was decreased (P <0.05) and the expression of Hu R protein in nucleus was increased (P <0.05). Conclusion PPS can induce apoptosis of T24 cells, which may be related to the role of Hu R in intracellular localization, reducing the stability of bcl-2 mRNA and decreasing the expression of bcl-2 protein.