The Influence of Nano-apatite on c-myc and p53 Gene in the Hepatocellular Carcinoma

The influence mechanism of the nano-apatite on the human hepatocellular carcinoma in vitro was investigated. Using the homogeneous precipitation method, the nano-apatite was synthesized at room temperature, and it was characterized with transmission electron microscopy (TEM) and the Zataplus. The influence on the expression of the c-myc and p53 gene in the human hepatocellalar carcinoma cell lines were tested with the TEM and hybridization in situ. The TEM and the Zataplus analyses show that the nano-apatite is distributed homogenously in size and needle-shaped sizes, which ranges from 67.5 nm to 88.3 nm. It is found that the nanoapatitet increases the volume of the human hepatocellalar carcinoma cells, makes extensive cytoplasmic vacuolization, the mitochondria swelling, chromatin in nucleus dispersed partially and condensed around the nuclear membranes. The interspace in nuclear membranes were separated and even the cytoplasm dissolved. It is also found that the expression of the c-myc gene is inhibited, but the p53 /s enhanced. The experimental results demonstrate that the nano-apatite enables the oneosis of the human hepatocellular carcinoma cells by down-regulation of the expression of the c-myc and up-regulation of the expression of the p53 in vitro.     

纳米磷灰石对肝癌癌基因表达的影响

研究纳米磷灰石体外对人肝癌细胞系Bel-7402的作用机理。应用均相共沉淀法室温下合成纳米磷灰石，用透射电镜和电位粒度仪进行表征；利用透射电镜、原位杂交细胞化学检测其对人肝癌细胞系Bel-7402 c-myc和p53基因表达的影响。纳米磷灰石呈均匀分散的针状颗粒．粒径范围在67．5～88．3nm之间。可使肝癌细胞体积增大，胞质空泡化，线粒体肿胀；部分细胞核内染色质分散，凝集在核膜周围，呈团块状，核膜间隙不等，胞浆溶解。抑制c-myc和增强p53基因的表达。纳米磷灰石在体外通过下调c-myc和上调p53基因的表达，致肝癌细胞胀亡。     

纳米羟基磷灰石的表征及其对肝癌抑制作用的研究

利用化学方法辅助超声制备出稳定单分散的羟基磷灰石(HAP)纳米粒子体系.应用原子力显微镜(AFM)、粒径电位仪对HAP纳米粒子进行表征;采用形态学观察和MTT比色法研究HAP纳米粒子对细胞系Bel-7402人肝癌细胞的作用;从单细胞荧光元素微区分析、超微结构及细胞周期的改变研究其作用机理.实验结果表明:HAP纳米粒子在体外对Bel-7402人肝癌细胞具有明显的抑制作用,通过进入到癌细胞内,阻滞细胞周期的进展,使细胞在G1期堆积,导致癌细胞胀亡.     

Morphological changes in mouse embryos cryopreserved by different techniques

Cryopreservation of mammalian embryos is an important tool for the application of reproductive biotechnologies. Subjective evaluation to determine embryo viability is often used. The determination of the best cryopreservation protocol depends on morphological and molecular analysis of cellular injuries. The main objective of this study was to compare two methods of cryopreservation by assessing morphological alterations of frozen embryos using light, fluorescence, and transmission electron microscope. Fresh (control), slow frozen, and vitrified mouse embryos were composed. To evaluate the viability of the embryos, the cell membrane integrity was assessed using Hoechst33342 and propidium iodide (H/PI) staining. Morphological analyses using hematoxylin and eosin (HE) staining were performed to test different techniques (in situ, paraffin, and historesin) by both light and fluorescence microscopy. Transmission electron microscope was used to detect ultrastructural alterations in Spurr- and Araldite-embedded samples. H/PI staining detected more membrane permeability in the vitrification (69.8%) than in the slow freezing (48.4%) or control (13.8%) groups (P < 0.001). Historesin-embedded samples showed to be more suitable for morphological analyses because cellular structures were better identified. Nuclear evaluation in historesin sections showed the induction of pycnosis in slow freezing and vitrification groups. Cytoplasm evaluation revealed a condensation and an increase in eosinophilic intensity (indicating apoptosis) in the slow freezing group, and weakly eosinophilic structures and degenerated cells (indicating oncosis) in the vitrification group (P < 0.05). Ultrastructural analyses confirmed HE morphological findings. It was concluded that both cryopreservation techniques resulted in oncosis and apoptosis injuries. However, vitrification caused more severe cellular alterations and reduced embryonic viability compared to slow freezing.     

The influence of nano-apatite on c -myc and p 53 gene in the hepatocellular carcinoma

The influence mechanism of the nano-apatite on the human hepatocellular carcinoma in vitro was investigated. Using the homogeneous precipitation method, the nano-apatite was synthesized at room temperature, and it was characterized with transmission electron microscopy (TEM) and the Zataplus. The influence on the expression of the c-myc and p53 gene in the human hepatocellular carcinoma cell lines were tested with the TEM and hybridization in situ. The TEM and the Zataplus analyses show that the nano-apatite is distributed homogenously in size and needle-shaped sizes, which ranges from 67.5 nm to 88.3 nm. It is found that the nanoapatitet increases the volume of the human hepatocellular carcinoma cells, makes extensive cytoplasmic vacuolization, the mitochondria swelling, chromatin, in nucleus dispersed partially and condensed around the nuclear membranes. The interspace in nuclear membranes were separated and even the cytoplasm dissolved. It is also found that the expression of the c-myc gene is inhibited, but the p53 is enhanced. The experimental results demonstrate that the nano-apatite enables the oncosis of the human hepatocellular carcinoma cells by down-regulation of the expression of the c-myc and up-regulation of the expression of the p53 in vitro. Funded by the State Key Laboratory of Advanced Technology for Materials Synthesis and Processing (Wuhan University of Technology, WUT2004Z02), and the Science and Technology Brain-storm Key Project of Hubei Province (2002AA105A)