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Objective To investigate the expression and clinical significance of lactate dehydrogenase A (LDHA) in breast cancer.Methods Tissue samples of 76 breast cancers and corresponding paired adjacent normal tissues were collected and made into tissue micrcarrays (TMAs).Immunohistochemistry (IHC) analysis was performed to detect the expression of LDHA and further analyzed the correlation of LDHA expression and clinicopathological variables and prognosis of breast cancers.Results LDHA was frequently upregulated in breast cancer tissues compared to the normal breast tissues (P ＜ 0.05).High LDHA expression was associated with distant metastasis (P ＜ 0.05) and worse patient prognosis (P ＜ 0.05).Conclusions LDHA is closely related to the occurrence and clinical progress of breast cancers.LDHA might be a potential novel molecular marker for diagnosis,prognosis and therapy in breast cancers.
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OBJECTIVE To investigate whether quinolinic acid(QA)induces autophagy in PC12 cells and its relationship with glycogen synthase kinase-3β(GSK-3β)/β-catenin related signaling path?ways. METHODS PC12 cells were treated with QA 2.5,5.0 and 10.0 mmol·L-1 for 24 h. The cell viability was determined by MTT assay. Autophagy fluorescent spots labelled form of microtubule-associated protein 1 light chain 3(LC3)was examined by LC3 immunostaining. The expressions of GSK-3β,β-catenin,LC3 and Beclin 1 were determined by Western blotting. RESULTS QA inhibited PC12 cell survival in a concentration-dependent manner,and IC50 was 8.7 mmol · L- 1. Compared with normal control group,QA 2.5,5.0 and 10.0 mmol · L-1 increased autophagic intracellular LC3 fluorescence spots,elevated the expression ratio of LC3-Ⅱ/LC3-Ⅰ and expression of Beclin 1 in PC12 cells(P<0.05). In addition,QA enhanced GSK-3βexpression and decreasedβ-catenin expression(P<0.05,P<0.01). CONCLUSION QA induces autophagy in PC12 cells. This mechanism may be associated with the activation of GSK-3β/β-catenin related signaling pathways.
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Aim To investigate the role of HIF-1 αin PC1 2 cell injury induced by quinolinic acid.Methods PC1 2 cells were treated with quinolinic acid at the do-ses of 2.5,5 and 1 0 mmol·L -1 ,the cell viability was determined by MTT reduction assay and LDH as-say,the intracellular levels of oxygen species was measured by assessing SOD and MDA levels,cell ap-optosis was determined by Hoechst 33258 staining,the intracellular distribution of HIF-1 αwas examined by HIF-1 αimmunostaining,and the expressions of HIF-1 α,Akt,p-Akt,Bcl-2 and Bax were determined by im-munoblotting analysis.Results Quinolinic acid in-duced cell injury in PC1 2 cells in a dose-dependent manner,and potentiated oxygen radical production and cell apoptosis.In addition,quinolinic acid enhanced HIF-1 αexpression and accumulation in nuclei.The p-Akt expression and Bax/Bcl-2 ratio was increased by quinolinc acid in PC1 2 cells.Conclusions HIF-1 αand Akt mediate qunolinc acid-induced cell apoptosis in PC1 2 cells.And cellular oxidative stress may con-tribute to the injury as well.
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A magnetic extraction sorbent based on Fe3 O4@poly ( methacrylic acid-co-ethylene dimethacrylate) ( Fe3 O4@MAED ) was synthesized using methacrylic acid and ethylene dimethacrylate as monomer and cross-linker, respectively. The magnetic sorbent was characterized by FTIR spectroscopy, elemental analysis and transmission electron microscopy. At the same time, the Fe3 O4@MAED was used to extract four benzoylurea pesticides in water and juice samples under magnetic dispersive solid phase microextraction ( MDSPME ) format. To obtain the optimal extraction conditions, several parameters, including the amount of sorbent, desorption solvent, extraction and desorption time, pH value, and ionic strength in sample matrix were investigated. Based on this, a fast, simple and sensitive method for the determination of benzoylurea pesticides in water and juice samples was developed by the combination of MDSPME with HPLC equipped with diode array detector. Under the optimized experimental conditions, the developed method possessed expected linear dynamic ranges, coefficients of correlation ( R2>0. 99 ) and sensitivity. The limits of detection (S/N=3) for target analytes were 0. 10-0. 19 μg/L in water and 0. 12-0. 30 μg/L in juice, respectively. The RSDs for intra-day were less than 7% and inter-day RSDs were less than 11%. The developed method was successfully applied to the determination of benzoylurea pesticides residues in water and juice samples and the recoveries of spiked target compounds were in the range of 69. 4%-118. 0%. The results demonstrated that the Fe3 O4@ MAED could extract benzoylurea pesticides effectively through multi-interactions including hydrophobic, hydrogen-bond and ion-exchange interactions between sorbent and analytes.
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Aim To investigate the expression and im-plication of HIF-1α, ROCK-2 , FoxM1 in PC12 cell in-jury induced by lead acetate. Methods PC12 cells were treated with lead acetate at the doses of 100 , 200 and 400 μmol·L-1 . The cell viability was determined by MTT reduction assay and LDH assay, the intracellu-lar production of oxygen species was measured by as-sessing SOD and MDA levels, cell apoptosis was deter-mined by Hoechst 33342 staining, the expressions of HIF-1α, ROCK-2 , FoxM1 , Bcl-2 and Bax were deter-mined by immunoblotting analysis. Results Lead ac-etate induced cell injury in PC12 cells in a dose-de-pendent manner, and it potentiated oxygen radical pro-duction and cell apoptosis. In addition, lead acetate enhanced HIF-1α and ROCK-2 expressions, increased Bax/Bcl-2 ratio and decreased FoxM1 expression. Conclusion Lead acetate can induce PC12 cell apop-tosis, which may be related with the expressions of HIF-1α, ROCK-2 and FoxM1 . Cellular oxidative stress may contribute to the injury as well.
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Aim To investigate the effects of stromal cell-derived factor-1α ( SDF-1α) on cell proliferation in primary cultured rat astrocytes and the possible mechanisms. Methods The primary cultured rat astr-cytes were treated with recombinant human SDF-1α at different concentrations, the cell proliferation was as-sessed by cell counting and 5-bromo-2’-deoxyuridine incorporation assay;intracellular calcium concentration was detected with calcium sensitive fluorescent probe;phorphorylation of extracellular regulated protein ki-nase1/2 ( ERK1/2 ) was determined by Western blot analysis;cell cycle transition was analyzed by flow cy-tometry analysis; mRNA expressions of cyclin A2 and cyclin B1 were determined by quantitative RT-PCR. Results Treatment of astrocytes with SDF-1α (5 -40 nmol·L-1 ) for 48 h induced significant cell prolifera-tion. SDF-1α at 20 nmol·L-1 increased the intracel-lular calcium concentration and the phosphorylation of ERK1/2. In addition, SDF-1α at 20 nmol·L-1 pro-moted the cell cycle transition from G0 to S and M pha-ses, and up-regulated the mRNA expressions of Cyclin A2 and Cyclin B1 . Conclusion SDF-1α significantly induces cell proliferation in primary cultured rat astro-cytes via enhancing calcium influx, ERK1/2 phospho-rylation, Cyclin expression and promoting cell cycle transition.
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A method for the determination of bisphenol A, octyphenol, nonylphenol in water samples was developed using stir bar sorptive extraction (SBSE) based on poly (vinylimidazole-divinylbenzene) monolithic material (SBSEM) combined with high performance liquid chromatography with diode array detection.To achieve the optimum extraction performance, several main extraction parameters, including extraction and desorption time, pH value and contents of inorganic salt in the sample matrix, were investigated.Under the optimized experimental conditions, the method showed good linearity and repeatability, low detection limits (S/N = 3) and quantification limits (S/N = 10) of the proposed method for the target compounds were achieved within the range of 0.13-0.66 and 0.44-2.19 μg/L, respectively.The extraction performance of SBSEM to the target compounds was also compared with commercial SBSE which used polydimethylsiloxane as coating.The proposed method was successfully applied to the determination of the target compounds in water samples.The recoveries of spiked target compounds in real samples ranged from 37.8%-101.1%.The results indicated that the developed method possessed advantages such as sensitivity, simplicity, low cost and high feasibility.
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Objective:To investigate the effect of exogenous nitric oxide(NO) on the growth and proliferation of gastric cancer cell line SGC-7901. Methods:The inhibitory effects of NO donor sodium nitroprusside (SNP) and nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methylester(L-NAME) on the proliferation of SGC-7901 cells were analyzed by MTT assay. The changes of mRNA and protein expression of proliferating cell nuclear antigen(PCNA) and caspase-3 were examined by RT-PCR and Western blotting. The cell cycle was measured using flow cytometry. Results:Compared with control group, more cells in the SNP group were arrested at G_1 and G_0 phases (P<0.05) and fewer cells were at S phases (P<0.05). SNP decreased the speed of cell-cycle progression from G_0/G_1 phase into S phase. SNP inhibited the proliferation of SGC-7901 cells and reduced the mRNA and protein expressions of PCNA and caspase-3. NOS inhibitor L-NAME reversed the effects of SNP. Conclusion:NO inhibited cell growth and proliferation, but accelerated apoptosis of gastric cancer cells.