0.05).Immunohistochemical staining for intratumoral aromatase in endometrial cancer was positive,21(42 %)in stromal cells.The expression of P450 aromatase in stromal cells correlated positively with advanced surgical stage and histologic grade,indepedent on muscular invasion and lymph node metastasis.Conclusion Aromatase P450 play an important function in establishment and devel- opment of the endometrioid endometrial cancer.The expression of P450 aromatase in intratumoral stromal cells correlated positively with the development and prognosis of endometrioid endometrial cancer.

0.05).The change of heart rate in 2 groups was greater than that of distilled water respectively(P

OBJECTIVETo observe the effect of Astragalus injection (AI) combined with chemotherapy on quality of life (QOF) in patients with advanced non-small cell lung caner (NSCLC).

METHODSSixty-NSCLC patients were randomly divided into the treated group (n = 30, treated with AI combined with chemotherapy) and the control group (n = 30, treated with chemotherapy alone). Chemotherapy of MVP protocol was applied to both groups. AI was supplemented to the treated group by intravenous dripping 60 ml per day. Treatment of 21-28 days as one treatment cycle, and 2-3 treatment cycles were applied.

RESULTSThe effective rate in the treated group was 40.0% and in the control group was 36.7%, the mean remission rate in the treated and control group was 5.4 months and 3.3 months, the median survival period 11 months and 7 months, and the 1-year survival rate 46.75% and 30.0%, respectively, the differences of these indexes between the two groups were all significant (P < 0.05). Moreover, the clinical improving rate and QOF elevation rate in the treated group was 80.4% and 43.3%, as compared with those in the control group (50.0% and 23.3% respectively), the difference was also significant (P < 0.01).

CONCLUSIONAI combined with chemotherapy can significantly improve the QOF in NSCLC patients of advanced stage.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Astragalus membranaceus ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Carcinoma, Squamous Cell ; drug therapy ; Cisplatin ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lung Neoplasms ; drug therapy ; Male ; Middle Aged ; Mitomycins ; therapeutic use ; Phytotherapy ; Quality of Life ; Vinblastine ; therapeutic use

OBJECTIVETo establish a heat shock protein 90alpha (HSP90alpha) expression-inhibited cell line and study the effect of lowered HSP90alpha level on cell stress response.

METHODSThe recombinant plasimid pSilencerHSP90 containing the 21nt small interfering RNA of human HSP90alpha was subcloned, purified and identified by DNA sequence analysis before introduced into mouse fibroblast cell line NIH-3T3 by electroporation. After G418 selection, the positive clones were identified by immunofluorescence and Western blotting. NIH-3T3 cells were subjected to hyperthermia at 44 degrees C for 40 min to simulate oxidative stress, and flow cytometry was performed to analyze the effect of low-level HSP90 on DNA damage under stress condition.

RESULTSImmunofluorescence and Westen blotting showed lowered HSP90 levels in the transfected cells. Compared with the control cells, cells subjected to hyperthermia displayed intensified DNA damage.

CONCLUSIONLow-level HSP90alpha causes the cells to be more vulnerable to oxidative stress condition, and HSP90 content can be associated with cell protection against such condition.

Animals ; Base Sequence ; Blotting, Western ; DNA Damage ; Fluorescent Antibody Technique ; HSP90 Heat-Shock Proteins ; genetics ; metabolism ; Hot Temperature ; Mice ; Models, Biological ; Molecular Sequence Data ; NIH 3T3 Cells ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

AIMTo establish a HSP90 highly expressing cell line and study the effect of high level of HSP90 on cell stress response.

METHODSThe recombined plasmid pSmycHSP, which contained the full length DNA coding for human HSP90 B, was introduced into mouse fibroblast cell line NIH-3T3 by electroporation after being subcloned, purified and identified by limited enzyme digestion. After screened by G418, the positive clones were selected and identified by immunocytochemistry and Western-blotting. Contrasted with NIH-3T3 cells transfected with empty plasmid, the effect of high-level HSP90 on cell proliferation and cell cycle was analyzed by MTT method and flow cytometry.

RESULTSThe rising level of HSP90 was shown by immunocytochemistry and Western-blotting. Compared with control, the growth of HSP90 highly expressing cell line slowed down and the DNA content of S phase was lower.

CONCLUSIONThe NIH-3T3 derived cell line, which stably expressed high level of HSP90 was established. The effect of high-level HSP90 on cell proliferation was to retard cell growth by affecting cell cycle.

Animals ; Cell Cycle ; Cell Proliferation ; HSP90 Heat-Shock Proteins ; metabolism ; Mice ; NIH 3T3 Cells

Objective To investigate expression of transforming growth factor-?1(TGF-?1)and connective tissue growth factor(CTGF) protein in renal tissues,and detect the levels of urinary TGF-?1 and CTGF in rats with diabetic nephropathy(DN).To observe the influence of losartan on expression of the two protein in renal tissue and excretion in urine.Methods Wistar rats were treated by intravenous injection of streptozotocin after right nephrectomy to induce DN rat model.The DN rats were randomly divided into two groups:DN experimental group and losartan treated group.The expression of TGF-?1 and CTGF in renal tissue were determined with immunohistochemical staining,urinary TGF-?1 and CTGF were assayed by enzyme-linked immunosorbent assay(ELISA) at 6,12 weeks respectively.Results Compared with losartan treated group,urinary protein excretion and the protein expression of TGF-?1 and CTGF significantly increased(P

AIMTo construct underexpression HSP90alpha and overexpression HSP90beta human hepatoma cell line HepG2.

METHODSThe combined plasimid pSilencerHSP90alpha and pSmycHSP90beta were introduced into HepG2 by electroporation, respectively. The result of transfection was identified by Western-blotting and the curve of cell growth was drew by MTT. Observe the cell vitality and expression of HSP90.

RESULTSExpression of HSP90 in transfected cell line was shown by Western-blotting: Compared with control, expression of HSP90 in the cells transfected with pSilencerHSP90alpha decreased, whereas that in the cells transfected with pSmycHSP90beta increased.The growth curves of the two groups of transfected cells was as the same as that of the control group.

CONCLUSIONThe stable overexpression HSP90beta and underexpression HSP90alpha HepG2 cell lines were established.

Base Sequence ; Electroporation ; HSP90 Heat-Shock Proteins ; genetics ; metabolism ; Hep G2 Cells ; metabolism ; Humans ; Molecular Sequence Data ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

Reduced radiosensitivity of lung cancer cells represents a pivotal obstacle in clinical oncology. The hypoxia-inducible factor (HIF)-1α plays a crucial role in radiosensitivity, but the detailed mechanisms remain elusive. A relationship has been suggested to exist between hypoxia and autophagy recently. In the current study, we studied the effect of hypoxia-induced autophagy on radioresistance in lung cancer cell lines. A549 and H1299 cells were cultured under normoxia or hypoxia, followed by irradiation at dosage ranging from 0 to 8 Gy. Clonogenic assay was performed to calculate surviving fraction. EGFP-LC3 plasmid was stably transfected into cells to monitor autophagic processes. Western blotting was used to evaluate the protein expression levels of HIF-1α, c-Jun, phosphorylated c-Jun, Beclin 1, LC3 and p62. The mRNA levels of Beclin 1 were detected by qRT-PCR. We found that under hypoxia, both A549 and H1299 cells were radio-resistant compared with normoxia. Hypoxia-induced elevated HIF-1α protein expression preferentially triggered autophagy, accompanied by LC3 induction, EGFP-LC3 puncta and p62 degradation. In the meantime, HIF-1α increased downstream c-Jun phosphorylation, which in turn upregulated Beclin 1 mRNA and protein expression. The upregulation of Beclin 1 expression, instead of HIF-1α, could be blocked by SP600125 (a specific inhibitor of c-Jun NH2-terminal kinase), followed by suppression of autophagy. Under hypoxia, combined treatment of irradiation and chloroquine (a potent autophagy inhibitor) significantly decreased the survival potential of lung cancer cells in vitro and in vivo. In conclusion, hypoxia-induced autophagy through evaluating Beclin1 expression may be considered as a target to reverse the radioresistance in cancer cells.

OBJECTIVE: To study the role of Novaferon on TNF-α production and expression of NF-κB mRNA in monocytes isolated from normal human peripheral blood and to provide theoretical basis for treatment of immunological diseases with Novaferon. METHODS: Monocytes were isolated from the peripheral blood in 30 healthy volunteers and divided into 5 groups: group A was blank control, group B was stimulated by LPS without Novaferon intervention, group C by LPS together with Novaferon intervention, group D by LPS before Novaferon intervention, which group E by LPS after Novaferon intervention. We detected the concentration of TNF-α after LPS stimulation and Novaferon intervention in the supernatant by ELISA and expression of NF-κB mRNA by RT-PCR. RESULTS: Novaferon inhibited TNF-α production by monocytes isolated from healthy volunteers induced by LPS in vitro in group D compared with group B [(1446.76±72.36) pg/mL vs (946.46±46.12) pg/mL, P<0.01], and the rate was 29.7%. There was no significant change in TNF-α concentration in group C and E compared with group B [(1446.76±72.36) pg/mL vs (1275.62±87.75) pg/mL, P>0.05; (1446.76±72.36) pg/mL vs (1383.62±86.96) pg/mL, P>0.05]. There was significant change in NF-κB mRNA expression in group D compared with group B (0.2829±0.0365 vs 0.4994±0.0604, P<0.01). There was no significant change in NF-κB mRNA expression in group C and group E compared with group B (0.4716±0.0616 vs 0.4994±0.0604, P>0.05; 0.4767±0.0600 vs 0.4994±0.0604, P>0.05). CONCLUSION Novaferon can suppress TNF-α secretion by monocytes induced by LPS in vitro, and it can affect the immunity function of monocytes, which may be associated with the downregulation of NF-κB mRNA expression in monocytes.
Humans ; Lipopolysaccharides ; pharmacology ; Monocytes ; drug effects ; metabolism ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; pharmacology ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors ; metabolism

OBJECTIVETo establish stress adaptation model of mouse fibroblast cell line NIH-3T3, to provide a group of parallel object for stress adaptation research, and to explore the function and mechanism of HSP90 in stress adaptation.

METHODSA stress-adapted cell model was established by thermal preconditioning (42 degrees C, 20 minutes), and the adaptation result was evaluated by observing the change of the membrane injury and the damage of DNA induced by the heat stress for the second time (44 degrees C, 20 minutes). The HSP90 content was detected by Western blot.

RESULTSAccording to the membrane injury and HSP90 synthesis induced by the heat stress for the second time, it was primarily confirmed that 6 hours after thermal preconditioning were the optimum stress protection time. When cells underwent heat stress for the second time 6 hours after thermal preconditioning, the membrane injury (15.4% +/- 2.6% vs 41.2% +/- 5.1%), damage of DNA (15.1% vs 26.3%) were decreased compared with the control group in which there was no preconditioning. The OD(HSP90)/OD(control) value indicated that the cellular HSP90 contents was decreased immediately after heat stress (44 degrees C, 40 min). The content of HSP90 was 0.82 +/- 0.18 in the heat stress group, 1.70 +/- 0.52 in the preconditioning group and 1.41 +/- 0.16 in the heat stress after preconditioning group.

CONCLUSIONWith the preconditioning for the NIH-3T3, the time point for the stress protection is confirmed, the model for the cellular stress adaptation is established and the protective effect of HSP90 is primarily confirmed in this model.

Adaptation, Physiological ; physiology ; Animals ; DNA Damage ; HSP90 Heat-Shock Proteins ; biosynthesis ; Heat Stress Disorders ; metabolism ; physiopathology ; L-Lactate Dehydrogenase ; metabolism ; Mice ; NIH 3T3 Cells