Oridonin up-regulates expression of P21 and induces autophagy and apoptosis in human prostate cancer cells.
Oridonin up-regulates expression of P21 and induces autophagy and apoptosis in human prostate cancer cells.
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DOI：
10.7150/ijbs.4554
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发表时间：
2012
影响因子：
9.2
通讯作者：
Li JC
中科院分区：
文献类型：
--
作者：
Li X;Li X;Wang J;Ye Z;Li JC
背景：冬凌草甲素（ORI）可抑制多种肿瘤细胞的增殖并诱导其凋亡。然而，该机制尚未完全理解。方法：体外培养人前列腺癌（HPC）细胞，CCK-8法检测细胞活力。透射电镜下观察超微结构变化。用吖啶橙子（AO）、MDC或DAPI化学染色检测细胞内酸性囊泡细胞器（AVO）和DNA的变化。Western Blot检测LC 3和P21的表达。流式细胞仪检测细胞凋亡率和细胞周期阻滞。结果如下：我们的研究表明，ORI处理后，人前列腺癌（HPC）细胞系PC-3和LNCaP的增殖抑制浓度和时间依赖性的方式。ORI诱导细胞周期阻滞在G2/M期。ORI处理24 h后，HPC细胞胞浆内出现大量双膜结构的自噬体和酸性泡状细胞器（AVO）。ORI导致LC 3-I转化为LC 3-II并将LC 3-II募集到自噬体膜。自噬抑制剂3-甲基腺嘌呤（3-MA）减少AVO的形成，并抑制LC 3-I向LC 3-II的转化。48 h时，ORI处理组细胞DNA断裂，染色质浓缩，表面微绒毛消失。ORI诱导凋亡细胞数量显著增加（PC-3：5.4%至27.0%，LNCaP：5.3%至31.0%）。通过营养饥饿促进自噬增加细胞活力，而通过3-MA抑制自噬促进细胞死亡。ORI可使P21蛋白表达增加，抑制自噬可完全逆转P21蛋白表达。结论：我们的研究结果表明，自噬发生在凋亡发生之前，并在ORI处理的HPC细胞中保护癌细胞。P21参与了ORI诱导的自噬和凋亡。本研究结果为深入了解ORI治疗前列腺癌的抗肿瘤机制提供了实验依据。
Background: Oridonin (ORI) could inhibit the proliferation and induce apoptosis in various cancer cell lines. However, the mechanism is not fully understood. Methods: Human prostate cancer (HPC) cells were cultured in vitro and cell viability was detected by the CCK-8 assay. The ultrastructure changes were observed under transmission electron microscope (TEM). Chemical staining with acridine orange (AO), MDC or DAPI was used to detect acidic vesicular organelles (AVOs) and alternation of DNA. Expression of LC3 and P21 was detected by Western Blot. Apoptotic rates and cell cycle arrest were detected by FACS. Results: Our study demonstrated that after ORI treatment, the proliferations of human prostate cancer (HPC) cell lines PC-3 and LNCaP were inhibited in a concentration and time-dependent manner. ORI induced cell cycle arrest at the G2/M phase. A large number of autophagosomes with double-membrane structure and acidic vesicular organelles (AVOs) were detected in the cytoplasm of HPC cells treated with ORI for 24 hours. ORI resulted in the conversion of LC3-I to LC3-II and recruitment of LC3-II to the autophagosomal membranes. Autophagy inhibitor 3-methyladenine (3-MA) reduced AVOs formation and inhibited LC3-I to LC3-II conversion. At 48 h, DNA fragmentation, chromatin condensation and disappearance of surface microvilli were detected in ORI-treated cells. ORI induced a significant increase in the number of apoptotic cells (PC-3: 5.4% to 27.0%, LNCaP: 5.3% to 31.0%). Promoting autophagy by nutrient starvation increased cell viability, while inhibition of autophagy by 3-MA promoted cell death. The expression of P21 was increased by ORI, which could be completely reversed by the inhibition of autophagy. Conclusions: Our findings indicated that autophagy occurred before the onset of apoptosis and protected cancer cells in ORI-treated HPC cells. P21 was involved in ORI-induced autophagy and apoptosis. Our results provide an experimental basis for understand the anti-tumor mechanism of ORI as treatment for prostate cancer.
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影响因子：
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