ABSTRACT
Objective To investigate the expression and clinical significance of myeloid cell leukemia-1 (MCL-1) and F-box and WD repeat domain-containing 7 (FBW7) in breast cancer polyploid induced by spindle poisons. Methods (1) Nocodazole spindle poison was used to treat breast cancer cell MDA-MB-231. The morphological changes of cells were ob?served under microscope, and cells were harvested in 0, 6, 12, 24, 48 and 72 h. The cell cycle and DNA-ploidy changes were examined by flow cytometry. The expressions of FBW7 and MCL-1 proteins were detected by Western blot assay. (2) A multikinase inhibitor (Sorafenib) with Nocodazole or Taxol was used to treat MDA-MB-231 cells. MCL-1 protein expression was detected by Western blot assay after 48 h treatment. The cell cycle and DNA-ploidy changes were examined by flow cy?tometry after 48 h treatment. MTT method was used to observe cell proliferation after 48 and 72 h treatment. Results (1)Af?ter treatment by Nocodazole, polyploid characteristics of large cell size and nucleus were appeared. The percentages of octa?ploid were (0.8±0.2)%, (8.5±2.3)%, (7.8±2.0)%, (9.9±0.9)%, (28.2±0.8)%and (35.1±4.9)%after 0, 6, 12, 24, 48 and 72 h treatment, showing the increasing trend in turn (P<0.001). The number of polyploidy (tetraploid and octaploid) cells was as high as (97.6±0.7)%after 48 h treatment. The expression level of FBW7 protein was decreased significantly but the expres?sion of MCL-1 protein was increased significantly after 48 h treatment. (2) After 48 h treatment, the expression level of MCL-1 protein, polyploidy percentage and cell proliferation decreased significantly in Nocodazole+Sorafenib group and Taxol+Sorafenib group compared with those of Nocodazole group and Taxol group (P<0.05). Conclusion The lower expression of FBW7 protein and over-expression of MCL-1 protein are correlated with the formation of breast cancer polyploidy. Sorafenib can reduce polyploid tumor cells by inhibiting MCL-1 protein expression.
ABSTRACT
Objective To study the effects of antisense oligodeoxynucleotides(ODN) of hEST2 (AODN) on telomerase activity and proliferation in ovarian cancer cell lines SKOV3 and COC1 Methods Antisense and sense human telomerse catalytic sub unit (hEST2) phosphorothioate (SODN)and random ODN were designed, synthesized and transfected into SKOV3 and COC1 cells by lipofectamine The expression of hEST2 mRNA and telomerase activity in SKOV3 and COC1 were tested by reverse transcription polymerase chain reaction and telomeric repeat amplification protocol before and after transfection The proliferation and growth in SKOV3 and COC1 were also investigated by methyl thiazolyl tetrazolium and growth curve before and after transfection Results AODN could down regulate the expression of hEST2 mRNA, inhibit telomerase activity and proliferation of ovarian cell lines The efficiency depends on dose and period of administration At 48 h, 30 ?mol/L AODN had the highest activity The expression of hEST2 mRNA were declined 54 6% and 44 6% in SKOV3 and COC1 respectively And also the inhibition of telomerase activity were 47 9% and 42 7% respectively in the two cell lines Conclusions AODN of hEST2 clearly inhibited the proliferation of ovarian cancer cell lines hEST2 may thus be a new target of gene therapy in ovarian carcinoma
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AIM: To observe the effect of altitude hypoxia on glutamate, aspartate and nitric oxide synthase (NOS) in the rat hypothalamus. METHODS: Using altitude hypoxia model, amino acid analysis system and the NADPH-d histochemistry, we determined the content of glutamate, aspartate and the number of NADPH-d neurons in the rat hypothalamus. RESULTS: After altitude hypoxia, the contents of glutamate, aspartate in the hypothalamus of rats were increased significantly, densely and deeply stained NADPH-d neurons were seen in hypothalamic paraventricular nucleus (PVN)and supraoptic nucleus(SON). If rat were pretreated with the NMDA receptor blockers Ketamine (ip,40 mg/Kg)or AP-V(i.c.v, 10 ?g) , the number of NADPH-d neurons in the rat hypothalamic PVN and SON was markedly less than that in corresponding altitude hypoxia group. CONCLUSION: NMDA receptor may take part in the expression of hypothalamic NOS induced by altitude hypoxia.
ABSTRACT
AIM:To observe the effect of altitude hypoxia on ?-aminobutyric (GABA) content and prepro-somatostatin mRNA (PPS-mRNA) in the rat hypothalamus. METHODS: Using altitude hypoxia model,in situ hybridization and amino acid analyzer, the number of PPS-mRNA and GABA content in rat hypothalamus was determined. RESULTS:After altitude hypoxia, the contents of GABA in hypothalamus and the number of PPS-mRNA neurons in periventricular nucleus (PeVN), paraventricular nucleus (PaVN) and arcuate nucleus (ArcN) increased significiantly. Bicuculline, a GABA receptor antagonits, could enhance PPS-mRNA expression evoked by altitude hypoxia, but had no effect on GABA content. CONCLUSION: Altitude hypoxia can induce neurotransmitter imbalance of hypothalamus.