Apogossypolone诱导前列腺癌PC-3细胞在体外的自噬

Apogossypolone Induces Autophagy of PC-3 Prostate Cancer Cells in vitro

目的研究ApoG2对前列腺癌PC-3细胞在体外的作用,了解其杀伤肿瘤细胞的机制。方法采用MTT法、吖啶橙染色、透射电镜、流式细胞技术、Western blot、免疫组织化学等方法观察了ApoG2对PC-3细胞的自噬与凋亡的诱导作用。结果 ApoG2可明显抑制PC-3细胞增殖;ApoG2作用于PC-3细胞72小时可诱导细胞自噬;加入自噬抑制剂3-MA可增强ApoG2诱导凋亡作用;ApoG2可以增强细胞内LC-3Ⅱ及Beclin-Ⅰ的表达,降低Bcl-2的表达水平。结论 ApoG2主要以诱导PC-3细胞发生自噬为主,抑制自噬可以促进凋亡的发生。

Objective To study the growth inhibitation action of gossypol derivatives,ApoG2 on the prostatic cancer PC-3 cells,and make the primary study about their anti-tumor mechanism.Methods The MTT analytical method,AO staining,transmission electron microscope,flow cytometer and Western blot for measuring the apoptosis related proteins were used to study the growth inhibition and induction of autophagy roles of ApoG2 on the prostatic cancer PC-3 cells in vitro.Results When the concentration of ApoG2 was higher than 2.5 μg/ml in solution,it had the obvious proliferation inhibition ability to the prostate cancer PC-3 cells in vitro,and it had the charcteristics of time and dose dependent.When ApoG2 was used for the treatment of PC-3 cell for 72 hours,the observation with AO staining and transmission electron microscope indicated that ApoG2 could obviously induce the autophagy of prostate cancer cells.The measurement results by the flow cytometer indicated that the apoptosis of prostate cancer cells induced by ApoG2 might be strengthened by the use of autophagic inhibitor 3-MA.The measurement of Bcl-2 expressions by the Western blot found that after ApoG2 at the concentration of 10.0 μg/ml was used for treatment of prostate cancer cells for 48 hours,the expression level of Bcl-2 decreased.The Beclin 1 and LC-3Ⅱexpressions in the tumor cells were obviously increased by immunohistochemistry method after ApoG2 treatment.Conclusion ApoG2 has the same obvious anti-proliferation role to the prostatic cancer PC-3 cells in vitro and its mechanism is to mainly induce the autophagy of tumor cells and to secondarily induce their death,the inhibition of autophagy may promote the occurrence of death.