1.Simvastatin inhibits expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 Cells
Guiqin LING ; Dongbo CHEN ; Baoqing WANG
Chinese Journal of Clinical Oncology 2013;(24):1523-1527
Objective:To investigate the effect of simvastatin on expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells. Methods:Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), immunofluo-rescent staining, flow cytometry, and Western blot were used to detect the expression of pluripotency markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells treated with different doses of simvastatin. Results:qRT-PCR revealed the more signifi-cant inhibition of gene expressions of Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells when subjected to high doses of simvastatin (10, 50, and 100 μmol/L) compared with the control group (P<0.05). By contrast, no significant difference was observed be-tween the expressions of low-dose simvastatin treatment (1 μmol/L) and control (P>0.05). The inhibitory effect of simvastatin on the gene expressions of Oct3/4 and Nanog was more significantly apparent at 50 and 100 μmol/L dosages than at 10 μmol/L (P<0.05). By contrast, no significant difference in the inhibitory expression of Sox-2 was observed among the three high-dose treatments (P>0.05). Between the two higher-dose treatments (50 and 100 μmol/L), no significant difference in the inhibitory expressions of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was found. Meanwhile, in the 10 μmol/L simvastatin treatment, immunoflurescent staining showed a marked reduction in the protein expression of all three pluripotent markers in MCF-7 cells, and flow cytometry demonstrated a decrease of Oct3/4-, Nanog-, and Sox-2-positive cells (P<0.05). Western blot further revealed that the protein expression of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was significantly declined by the same simvastatin dose (P<0.05). Conclusion: Simvastatin can inhibit the expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells, proving the anti-cancer properties of simvastatin.
2.Gene expression profile in mice model of experimental autoimmune encephalomyelitis using cDNA microarrays
Guiqin SONG ; Weiping WU ; Chunjie SONG ; Ling YIN ; Ke ZHU
Chinese Journal of Neurology 2001;0(01):-
Objective To construct the genes expression profile database of experimental autoimmune encephalomyelitis (EAE) mice model and to screen the high candidate genes with microarray analysis. Methods Polymerase chain reaction (PCR) products on behalf of 4 096 genes from mixed mice tissue library were assembled on the modified Oako glass slide. The general RNA from 6 EAE mice model and 6 normal mice head was transcripted into cDNA by RT-PCR and labelled with cy-3 and cy-5.6 groups each consist one EAE mice and a control. These two probes were mixed and hybridized with the above mentioned gene chips. Scan array 4000 was used for scanning the hybridizing signals and GenePixPro 3.0 for date analysis. Results Of the total 4 096 genes monitored, 43 genes in group one showed changes in expression level of the ratio outside 0.5 and 2, 30 genes differed in group two, 176 genes changed in group three, 76 in group four, 294 in group five and 129 in group six. Conclusions The results showed the consistent different genes expression throughout the EAE mice model. The genes related to immune, cell structure, cell cycle, ion channel, signal transduction, protein synthesis and metabolism are involved in EAE pathogenesis. The results provide deeper insights into the mechanism of EAE and multiple sclerosis.