Acute hypoxia induces apoptosis in serum-deprived prostate cancer LNCaP cells
Acute hypoxia induces apoptosis in serum-deprived prostate cancer LNCaP cells
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急性缺氧诱导去血清前列腺癌LNCaP细胞凋亡
DOI：
10.21037/tcr.2017.10.13
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发表时间：
2017-12-01
影响因子：
0.9
通讯作者：
Chen, Yaqing
中科院分区：
文献类型：
--
作者：
Liu, Weiyong;Zhu, Yunkai;Chen, Yaqing
背景：人前列腺癌LNCaP细胞可以建立一种雄激素敏感的前列腺癌动物模型，该模型与临床疾病非常相似。然而，当皮下异种移植到免疫缺陷小鼠中时，LNCaP细胞的植入产生低摄取率。鉴于LNCaP细胞是敏感的缺氧，我们假设，LNCaP细胞可能会发生大量的凋亡，在急性缺氧条件下的serum-deprivation.Methods：血清剥夺LNCaP细胞培养和暴露于常氧（21.0%O-2）或缺氧（1.0%O-2）条件。荧光显微镜下Hoechst 33258染色观察细胞核形态学变化。分别用CCK-8试剂盒和伤口愈合试验检测细胞活力和迁移能力。细胞内活性氧（ROS）的产生通过荧光探针测定来测量。结果：急性缺氧条件下，去血清LNCaP细胞倾向于形成多细胞聚集体，细胞活力和迁移能力明显受到抑制。急性低氧暴露24小时诱导显著的ROS产生，其在细胞间分布不均匀。流式细胞仪分析显示，LNCaP细胞在急性缺氧条件下凋亡呈时间依赖性增加。此外，急性缺氧诱导的LNCaP细胞的Δ Psi m的崩溃，它也增加了细胞周期阻滞在G 0/G1期在时间依赖martens.Conclusions：急性缺氧诱导大量的细胞凋亡LNCaP细胞在血清剥夺条件下。本研究结果为LNCaP裸鼠皮下移植瘤成瘤方法的改进提供了有价值的信息。
Background: Human prostate cancer LNCaP cells can develop an androgen-sensitive prostate cancer animal model that closely mimic clinical disease. However, implantation of LNCaP cells yields a low take rate when subcutaneously xenografted in immunodeficient mice. Given that LNCaP cells are sensitive to hypoxia, we hypothesized that LNCaP cells might undergo substantial apoptosis in response to acute hypoxia under conditions of serum deprivation.Methods: Serum-deprived LNCaP cells were cultured and exposed to either normoxic (21.0% O-2) or hypoxic (1.0% O-2) conditions. Morphological changes in nuclei were evaluated with Hoechst 33258 staining under fluorescence microscopy. Cell viability and migration were assayed by Cell Counting Kit-8 (CCK-8) and wound healing assay, respectively. The production of intracellular reactive oxygen species (ROS) was measured by a fluorescent probe assay. Apoptosis, mitochondrial membrane potential (Delta Psi m), and the cell cycle of LNCaP cells were analyzed by flow cytometry.Results: Serum-deprived LNCaP cells tended to form multicellular aggregates, and the viability and migration of LNCaP cells were markedly suppressed under acute hypoxia. Acute hypoxia exposure for 24 hours induced significant ROS production, which was distributed heterogeneously among cells. LNCaP cells exhibited a time-dependent increase in apoptosis under acute hypoxia by flow cytometric analysis. Furthermore, acute hypoxia induced the collapse of the Delta Psi m of LNCaP cells, and it also increased cell cycle arrest at the G0/G1 phases in a time-dependent manner.Conclusions: Acute hypoxia induces substantial apoptosis in LNCaP cells under conditions of serum deprivation. The findings may provide valuable information for improving the tumor-formation method of LNCaP subcutaneous xenografts in nude mice.