Effects of novel genistein derivative 5-hydroxy-4'-nitro-7-propionyloxyisoflavone on proliferation and apoptosis of human cervical cancer HeLa cells in vitro
10.3781/j.issn.1000-7431.2017.11.269
- Author:
Aiqiong TANG
1
Author Information
1. First Department of Gynecology, Maternal and Child Health Hospital of Hunan Province
- Publication Type:Journal Article
- Keywords:
5-Hydroxy-4'-nitro-7-propionyloxy-isoflavon;
Apoptosis;
Cell proliferation;
Uterine cervical neoplasms
- From:
Tumor
2017;37(9):924-935
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effects of 5-hydroxy-4'-nitro-7-propionyloxy-isoflavone (HNPI), a new derivative of genistein, on proliferation and apoptosis of human cervical cancer HeLa cells in vitro , and to explore the possible molecular mechanism. Methods: HeLa cells were treated with different concentrations of HNPI (5, 10, 20, 40, 60 and 80 μmol/L) for 48 h, while the cells were treated with 0.9% sodium chloride solution as the control group. The proliferation inhibitory rate of HeLa cells was detected by MTT assay. The alterations of HeLa cells on surface morphology, height, width, root-mean-squared roughness (Rq) and average roughness (Ra) were measured by atomic force microscopy. The changes of nuclear morphology of HeLa cells stained with DAPI were detected by laser scanning confocal microscopy. The apoptosis rate, cell cycle distribution, the level of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) in HeLa cells were detected by FCM. Results: The proliferation of HeLa cells was significantly inhibited by HNPI in a dosedependent manner, and there was statistical significance as compared with the control group when the concentration of HNPI was greater than 10 μmol/L (all P < 0.05). The value of half maximal inhibitory concentration (IC50) of HNPI on HeLa cells was 46.83 μmol/L. After treatment with 20 and 60 μmol/L HNPI for 48 h, HeLa cells became round and shrinkage, the cytoplasm condensed, the surface roughness decreased, the filopodia structure was shrunk and even completely destroyed; the surface ultrastructure of HeLa cells became smooth and homogeneous, and merged into large protuberance or bulk; the height of single cell improved, but the width, Rq and Ra of cells down-regulated in a dose-dependent manner as compared with the control group (all P < 0.05); the nucleus indented, the chromosomes gathered, and the apoptosis bodies formed. Furthermore, the apoptosis of Hela cells was induced and the cell cycle was arrested at G0/G1 phase by HNPI in a dose-dependent manner, and there were statistical differences as compared with the control group when the concentration of HNPI was greater than 10 μmol/L (all P < 0.05). Meanwhile, the level of intracellular ROS in HeLa cells was increased, and MMP was decreased in a dose-dependent manner as compared with the control group (both P < 0.05). Conclusion: HNPI can significantly inhibit the proliferation and induce the apoptosis of HeLa cells in vitro , which may be related to the cell damage and ultrastructure change caused by HNPI, resulting in accumulation of intracellular ROS, reduction of MMP, and the blockage of cell cycle at G0/G1 phase.
