Role of allograft inflammatory factor-1 in regulating the proliferation, migration and apoptosis of colorectal cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of allograft inflammatory factor-1 (AIF-1) in colorectal cancer (CRC) progression and explore the possible mechanism.</p><p><b>METHODS</b>The expression levels of AIF-1 in 70 CRC tissues and paired adjacent tissues were detected using immunohistochemistry and Western blotting, and the correlation of AIF-1 expression with the clinicopathological features of the patients was analyzed. In the CRC cell line SW480, the functional role of AIF-1 in regulating tumor progression was investigated by transfecting the cells with an AIF-1-overexpressing plasmid (AIF-1) and a negative control plasmid (NC). EdU proliferation assay and flow cytometry were used to assess the cell proliferation and cell cycle changes; Transwell migration assay and Annexin V-APC/7-AAD apoptosis assay kit were used to analyze the cell migration and apoptosis. The changes in the biological behaviors of the cells were observed after application of SB203580 to block the p38 MAPK pathway. The expression levels of CDK4, cyclin D1, P21, P27, MMP2, MMP9, Bax, Bcl2, Bcl-xl, p38 and p-p38 were detected using Western blotting.</p><p><b>RESULTS</b>AIF-1 was down-regulated in CRC tissues compared with the adjacent normal tissues, and its expression level was positively correlated with lymph node metastasis (P=0.008), TNM stage (P=0.003) and tumor size (P=0.023). Overexpression of AIF-1 in SW480 cells significantly reduced EdU-positive cells and caused obvious cell cycle arrest in G1 phase (P<0.05). AIF-1 overexpression resulted in significantly lowered protein expressions of CDK4 and cyclin D1, enhanced expressions of P21 and P27, attenuated cell migration ability (P<0.001), and decreased protein levels of MMP2 and MMP9. AIF-1 overexpression also induced obvious apoptosis of SW480 cells (P<0.01), significantly increased the protein levels of Bax and p-p38, and decreased the protein levels of Bcl-2 and Bcl-xl; SB203580 significantly attenuated the apoptosis-inducing effect of AIF-1 overexpression.</p><p><b>CONCLUSION</b>AIF-1 plays the role of a tumor suppressor in CRC by inhibiting cell proliferation, suppressing cell migration and inducing cell apoptosis. AIF-1 overexpression promotes the apoptosis of CRC cells by activating the p38 MAPK pathway.</p>

Effect of quercetin on gene expression in glucose-oxygen deprived astrocytes / 中华神经医学杂志

Objective To study the effect of quercetin on gene expression in glucose-oxygen deprived astrocytes using large-scale oligo microarray technology.Methods Astrocytes were primarily cultured in vitro and divided into ischemia and hypoxia group and ischemia and hypoxia plus quercetin treatment(50 μmol/L)group; ischemia and hypoxia cells from these 2 groups were induced by anaerobic culture for 4 h,and then,the nutrient solutions were added into each group,respectively.Twenty-four h after that,cDNA microarray was employed to select the differentially expressed genes in the 2 groups,and these genes were confirmed by quantitative RT-PCR.Results The cDNA microarray indicated that the expressions of 180 genes had significant changes at the mRNA level between ischernia and hypoxia group and ischemia and hypoxia plus quercetin treatment group,of which 49 genes were up-regulated and 131 were down-regulated.One hundred and forty-eight differentially expressed genes were confirmed by RT-PCR,including 34 up-regulated genes and 114 down-regulated genes,which showed that 82.2％(148/180)genes that matched with the results of cDNA microarray.Conclusion Gene expression profiling by large-scale oligo microarray provides good understanding of the molecular mechanism of quercetin in glucose-oxygen deprived astrocytes,and laids the foundation for investigating the influence of quercetin and astrocytes in hypoxic-ischemic brain damage.

Effect of quercetin on neural stem cell proliferation in the subventricular zone of rats after focal cerebral ischemia-reperfusion injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of quercetin on the proliferation of neural stem cells in the subventricular zone (SVZ) of rats after focal cerebral ischemia.</p><p><b>METHODS</b>An adult rat model of middle cerebral artery occlusion (MCAO) model was established by placement of an intraluminal filament at the origin of the MCA. Quercetin was administered intraperitoneally in the rats at a dose of 50 mg/kg every 3 days starting at 6 h after MCAO, and BrdU (50 mg/kg daily) was also injected intraperitoneally starting at 4 h after MCAO. BrdU-positive cells in the SVZ were counted at 7, 14 and 21 days after MCAO.</p><p><b>RESULTS</b>Compared with the sham-operated group, the rats in the ischemic model group showed significantly increased BrdU-positive cells in the ipsilateral SVZ 7 days after MCAO, reaching the peak level on day 14 and beginning to decrease on day 21 (P<0.05). The number of ipsilateral BrdU-positive cells in quercetin group was significantly greater than that in the model group on days 7, 14 and 21 (P<0.05), and maintained the high level on day 21.</p><p><b>CONCLUSION</b>Quercetin can maintain a high level of neural stem cell proliferation in the SVZ after focal cerebral ischemia in adult rats.</p>

Protective effect of mild hypothermia on astrocytes with traumatic or ischemic injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effect of mild hypothermia on rat astrocytes with traumatic or ischemic injury.</p><p><b>METHODS</b>Rat astrocytes in primary culture were subjected to scratching or hypoxic injury and exposed to normothermia (37 degrees celsius;) or hypothermia (34 or 32 degrees celsius;) for 24 h. The morphology of the astrocytes was evaluated by live/dead staining, and the cell injury was measured by lactate dehydrogenase (LDH) release assay.</p><p><b>RESULTS</b>As the temperature reduced the LDH release rate from the cells in hypoxic group decreased significantly, to (11.48 - or + 1.53)% at 34 degrees celsius; and (3.79 - or + 0.45)% at 32 degrees celsius; as compared to that in normothermia [(33.02 - or + 3.58)%] in the absence of rat white blood cells (WBC) (P<0.001). LDH release rate of the hypoxic cells further decreased in the presence of rat WBC to (51.14 - or + 2.17 )% at 37 degrees celsius;, (19.53 - or + 4.37)% at 34 degrees celsius; and (16.68 - or + 1.47)% at 32 degrees celsius; (P<0.001). In the scratched cells, with or without WBC, LDH release rate showed no significant variation between the 3 temperatures (P>0.05).</p><p><b>CONCLUSION</b>Mild hypothermia offers obvious protective effects on rat astrocytes against ischemic damage but not against mechanical injury.</p>

Construction of delta-pIRES2-EGFP plasmid and its expression in HEK293 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the delta-pIRES2-EGFP plasmid and investigate its expression in HEK293 cells.</p><p><b>METHODS</b>Full length cDNA of rat delta opioid receptor gene amplified from rat brain tissues using reverse transcription and nested PCR was cloned into pMD20 T vector. The delta cDNA was inserted into pIRES2-EGFP plasmid to construct the recombinant eukaryotic plasmid delta-pIRES2-EGFP, which was transfected into HEK293 cells via Lipofectamine2000. The expression of delta was examined under fluorescence microscope.</p><p><b>RESULTS</b>The recombinant delta-pIRES2-EGFP plasmid was successfully constructed, and high expression of delta was detected in HEK293 cells transfected by the plasmid.</p><p><b>CONCLUSION</b>delta-pIRES2-EGFP has been successfully cloned, which shows high expression of delta in HEK293 cells.</p>