| Aim: To study the response mechanism of cancer cell cycle regulation through liver microsomal lipid peroxide models and transplanted tumor-bearing animals such as rats to compare the actions among synchronization, antioxidation and γ-irradiation on transplanted tumor-bearing rats.Methods: The liver microsome of rat was prepared by Slater' s assay, liver microsomal lipid peroxide models including models stimulated by carbon tetrachloride(CCl4), cumene hydroperoxide(CHP) and vitamin C (Vc)/Iron(Fe2+) to estimate the capacity and reactivity on free radicals of antioxidants in which the Lingzhi sporule powder and pollen from tea were tested. The transplanted tumor-bearing rats injected by Walker 256 mammary cancer were used to compare the actions among synchronization, antioxidation and γ-irradiation and to study the response mechanism of cancer cell cycle regulation. The animals were divided into two groups: control group (10 rats) and experimental group(180 rats). All the transplanted tumor-bearing rats were divided into six groups: con-group (control group), pos-group (tumor-bearing animal group, positive group), syn-group (synchronization animal group induced by Lingzhi sporule powder, synchronization group),ant-group (antioxidationanimal group), irr-group (irradiation group), irr+ani-group(irradiation plus antioxidation group) and con+irr+ani-group(irradiation plus antioxidation plus synchronization group), with30 rats in each group. Some experimental transplanted tumor-bearinganimal groups including irr-group, irr+ani- group and con+irr+ani-group received total 60 Gy of 60CO- γ ray irradiation in ten days.The animal growth status, the livability, the weight of tumor, thehaematological index, the cell microgel electrophoresis (DNAinjury index), the microstructure of cancer cell, the immunolo-gical index, the regulator of cell cycle and differential gene weredetermined.Results:1. The transplanted tumor-bearing animal model injected by Walker 256 mammary cancer were successfully duplicated, which may provide technique guarantees for related research in this field.2. The andioxidative activity and dose-effect relation of Lingzhi sporule powder on models stimulated by carbon tetrachloride (CC14), cumene hydroperoxide (CHP) and vitamin C(Vc) /Iron(Fe2+) were confirmed. The Lingzhi sporule powder had strong synchronization of 91. 7% cancer cells at G1 phase on arresting. Cell cycle increased sensitivity.3. Pollen had certain antioxidation on peripheral blood, liver and bone marrow. It enhanced the kill effect on tumor cells and at the same time eliminated the side effect of free radical.4. The experiment further demonstrated that the main mechanism of local radiation's side effect was free radical damage.5. Antioxidants facilitated the up-regulation of the expression of CDK4, E2F-I, P2t and P53, but down-regulation of the expression of Rb. Synchronization down-regulated the expression of Cyclin D, and Rb but up-regulated Cyclin E and E2F-1. Irradiation slightly up-regulated all but Rb.Antioxidants had some significant control effects on the gene change caused by tumor-bearing and irradiation.0 In all the irr-groups, the leucocyte number degraded significantly (P<0. 05). The number of leucocyte and erythrocyte in con+irr+ani-group was higher than the synchronization' s. The antioxidants facilitated the leucocyte to step up in some condition. Meanwhile, the synchronization and antioxidant protected the injury of platelet to some extent.?Local irradiation on tumor-bearing animals, the weight of tumor in order was: irr-group>syn-group>irr+anigroup. Antioxidants had the best inhibitive effect on tumor.?Histology ultramicrostructure indicateded that in con+irr+ani-group the tumor cells showed karyopycnosis, pykno-chrom-atin and higher-electrode-density microbody, which was typical sign of apoptosis.@In the groups that were added antioxidants, the activity of GPT went down instead of went up. Therefore, the antioxidants had some apparent protective effect on cellular membrane.? Antioxidants significantly increased the ratio of CD4VCD8+T, which showed that synchronization plus antioxidation could enhance the immunocell function.?The outcome of content determination of MDA, GSSG, GSH in liver homogenate after different treatment showed that the MDA level of liver in syn-group increased notably, anti-group had inhibitive effect on MDA level, the content of GSH in tumor-bearing animals decreased significantly, but after syn+anti treatment, the level of GSH in liver stepped up.6. Using the gene-chip triage techniques, the up-and down-regulation of differential gene under different treatments was as follows.(1) There were 4 genes which might up-regulate in irradiation,antioxidants, and down-regulate in antioxidants/irradiation: Aminolevulinic acid synthase 1, Carbonic anhydrase 2, Glycera-Dehyde-3-phosphate dehydrogenase and Ribosomal protein L41.(2) There were 14 genes which might up-regulate in both irradiation and antioxidants: Annexin 1, CDdo, Fos like antigen 2, Golgi SNAP receptor complex member 1, Hemoglobin beta, Lysozyme, Procollagen, type I alpha 1, Ribosomal protein, Proteoglycan peptide core protein L5, Short form of beta II spectrin, Tropomyosin 3, Solute carrier family 25, Synaptogyrin 1 and Thymosin beta-4.( 3) There were 16 genes which might down-regulate in irradiation and turn back in antioxidants/irradiation: Acid phosphatase 5, tartrate resistant, Cyclic AMP phosphor protein, Dynein, Ferritin subunit H, mitochondrial dicarboxylate carrier, liver nuclear proteinP4-, Pancreatic and duodenal homeobox gene 1, Prothymosin alpha, proteolipid protein, Ribosomal protein S3a, Ribosomal protein L37, Sodium channel, voltage-gated, type IPot-assium voltage-gated channel, subfami, high mobility group box 2.(4)There were 5 genes which might up-regulate in irradiation: Heat shock 70kD protein 8, Methyl CpG binding protein 2, Prolactin receptor, IFP, Vimentin, submandibular gland secretory Glx-rich protein.(5) There were 5 genes which might up-regulate in antioxidants: B-cell translocation gene 2, Carbonyl reductase, GATA-binding protein 1 , Mast cell protease 8 and PMF31.(6) There was one up-regulated mighty gene in antioxidants: Neurogranin.(7) There were 2 genes which might up-regulate in irradiation and turn back in antioxidants/irradiation: Tumor protein and F-0-ATPase subunit b mRNA.(8) There was one up-regulated gene in irradiation and one mighty down-regulated gene in antioxidants: Transferrin.(9) There were 2 genes which might down-regulate in irradiation and mighty up-regulated gene in antioxidants: Ig active Iambda2-like chain mRNA and Kirsten rat sarcoma viral oncogene homol.(10) There were 2 genes which might down-regulate in irradiation and mighty down-regulated gene in antioxidants: defensin RatNP-3 precursor mRNA and S100 calcium-binding protein A9.(11) There were 4 down-regulated mighty genes in antioxidants: Ribosomal protein Li2, S100 calcium-binding protein A9, Synaptic vesicle glycoprotein 2b and mitochondrial ATP synthase beta subunit.Conclusion:1. The transplanted tumor-bearing animal models injected by Walker 256 mammary cancer were successfully duplicated.2. Lingzhi sporule powder had andioxidative activity and dose-effect relation. It also had strong synchronization (more than 90%).3. Pollen had certain andioxidative activity and could eliminate the side effect of free radical on peripheral blood, liver and bone marrow.4. The experiment further proved that the main mechanism of local radiation' s side effect was free radical damage. It provided necessary ground treatment for cancer using multi-functional antioxidants to diminish the side effect.5. Antioxidants could facilitate the up-regulation of the expression of CD&, E2F1, P27 and P53 but down-regulation of the expression of Rb. Synchronization down-regulated the expression of Cyclin Dl and Rb but up-regulated Cyclin E and E2F1. Irradiation slightly up-regulated all but Rb.6. Some differential genes were found. (1) There were 2 genes which up-regulated in irradiation and turned back in... |
PDF Full Text Request