目的 探究棕矢车菊素调节程序性死亡配体-1（PD-L1）表达对前列腺癌细胞增殖、凋亡和免疫逃逸的影响。方法 培养人正常前列腺上皮细胞RWPE-1和前列腺癌细胞PC-3，CCK-8法检测不同浓度（0、5、10、20、40、80 μmol/L）棕矢车菊素对RWPE-1和PC-3细胞活力的影响。PC-3细胞分为对照组、棕矢车菊素（20、40 μmol/L）组、多西他赛组、棕矢车菊素＋重组PD-L1组。通过集落形成、Annexin V/PI双染法和免疫印迹法检测细胞增殖、凋亡及蛋白表达；流式细胞仪检测共培养体系中PC-3细胞凋亡；ELISA检测共培养上清中γ干扰素、颗粒酶B含量。结果 棕矢车菊素≥20 mmol/L时PC-3细胞活力显著降低（P＜0.05）。与对照组比较，棕矢车菊素各浓度组PC-3细胞集落形成数量，c-myc、增殖细胞核抗原（PCNA）、X连锁凋亡抑制蛋白（XIAP）、PD-L1、淋巴细胞激活基因3（LAG-3）、B7同源物（B7-H3）蛋白表达均显著降低，PC-3细胞凋亡率、C-多聚ADP核糖聚合酶（PARP）蛋白表达以及共培养体系中PC-3细胞凋亡率、γ干扰素、颗粒酶B含量均显著升高（P＜0.05）；与棕矢车菊素40 mmol/L组比较，棕矢车菊素＋重组PD-L1组PC-3细胞集落形成数量，c-myc、PCNA、XIAP、PD-L1、LAG-3、B7-H3蛋白表达均显著升高，PC-3细胞凋亡率、C-PARP蛋白表达以及共培养体系中PC-3细胞凋亡率、γ干扰素、颗粒酶B含量均显著降低（P＜0.05）。结论 棕矢车菊素通过下调PD-L1等免疫检查点表达抑制前列腺癌细胞增殖及免疫逃逸，促进凋亡。

Objective To explore the effects of jaceosidin on the proliferation, apoptosis and immune escape of prostate cancer cells by regulating PD-L1 expression. Methods Human normal prostate epithelial cells RWPE-1 and prostate cancer cells PC-3 were cultured. Effects of different concentrations (0, 5, 10, 20, 40, and 80 μmol/L) of jaceosidin on the viability of RWPE-1 and PC-3 cells were measured by the CCK-8 method. PC-3 cells were assigned into the control group, the jaceosidin (20 and 40 μmol/L) group, docetaxel group, and jaceosidin + recombinant PD-L1 group. Colony formation, Annexin V/PI double staining and Western blotting were used to detect cell proliferation, apoptosis and protein expression. The apoptosis of PC-3 cells in the co-culture system was detected by flow cytometry. ELISA was used to measure the levels of IFN-γ and granzyme B in the co-culture supernatant. Results When jaceosidin ≥ 20 mol/L, it has an inhibitory effect on the viability of PC-3 cells (P < 0.05). Compared with the control group, the colony formation quantity, and the protein expression of c-myc, PCNA, XIAP, PD-L1, LAG-3, and B7-H3 of PC-3 cells were decreased in the jaceosidin group, but the apoptosis rate of PC-3 cell, the protein expression of C-PARP, and the apoptosis rate of PC-3 cells, interferon-γ and granzyme B content in the co-culture system were higher (P < 0.05). Compared with the jaceosidin 40 μmol/L group, the colony formation quantity, protein expression of c-myc, PCNA, XIAP, PD-L1, LAG-3, and B7-H3 of PC-3 cells in the jaceosidin + recombinant PD-L1 group were higher, but the apoptosis rate, C-PARP protein expression, and the apoptosis rate of PC-3 cells, interferon-γ and granzyme B content in the co-culture system were lower (P < 0.05). Conclusion Jaceosidin inhibits the proliferation and immune escape of prostate cancer cells and promotes apoptosis by down-regulating the expression of immune checkpoints such as PD-L1.

R287.3

遵义市科技计划项目[遵市科合HZ字（2024）20号]