Effect of 5-aza-2'-deoxycytidine on growth and methylation of RUNX3 gene in human pancreatic cancer cell line MiaPaca2 / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effect of demethylating agent 5-aza-2'-deoxycytidine (5-Aza-CdR) on the growth of human pancreatic cancer cell line MiaPaca2 and the expression and methylation of tumor suppressor gene RUNX3.</p><p><b>METHODS</b>Human pancreatic cancer cell line MiaPaca2 cells were treated with different concentrations of 5-Aza-CdR. Morphological changes of MiaPaca2 cells were observed by light microscopy. The activity of cell proliferation was analyzed by MTT assay. The changes of RUNX3 mRNA expression were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Changes of RUNX3 gene methylation was detected by methylation-specific polymerase chain reaction.</p><p><b>RESULTS</b>MiaPaca2 cells were treated with 2.5, 5, 10 and 20 µmo1/L 5-Aza-CdR, respectively. The inhibition rates of MiaPaca2 cells treated for 24 h were (9.17 ± 2.15)%, (10.75 ± 2.04)%, (12.57 ± 1.64)% and (18.70 ± 1.51)%, respectively. The inhibition rates were (14.94 ± 1.68)%, (18.60 ± 1.57)%, (22.84 ± 1.58)% and (33.24 ± 1.53)%, respectively, after 48 h treatment; (21.46 ± 1.60)%, (28.62 ± 1.72)%, (35.14 ± 1.64)% and (45.06 ± 1.47)%, respectively, after 72 h treatment; and (26.35 ± 1.71)%, (34.48 ± 1.69)%, (40.05 ± 1.60)% and (49.99 ± 1.61)%, respectively, after 96 h treatment. The differences between inhibition rates of each experimental and control groups (0.00 ± 0.00)% were statistically significant (P < 0.05). At the same time, the inhibition rates of different concentration groups showed significant differences (P < 0.05). At 48 h, 72 h and 96 h, the inhibition rates of each pair concentration groups showed significant differences (P < 0.05). 5-Aza- CdR inhibited the growth of MiaPaca2 cells, and the higher the concentration, the stronger the inhibition after 24 h. 5-Aza-CdR also reversed the methylation status of RUNX3 gene, and restored the expression of RUNX3 mRNA with a dose-effect relationship.</p><p><b>CONCLUSIONS</b>The methylation of RUNX3 gene is significantly related with the occurrence and development of pancreatic cancer, and abnormal methylation of RUNX3 gene may contribute to the loss of RUNX3 mRNA expression. 5-Aza-CdR may effectively cause reversion of RUNX3 methylation, and treatment with 5-Aza-CdR can reactivate the gene expression and inhibit the cell growth. This may provide a new way for diagnosis and treatment of pancreatic cancer.</p>

Identification of gastric cancer-related genes by multiple high throughput analysis and data mining / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate gastric cancer-related genes by combined multiple high throughput analysis and data mining, and to further identify gene markers that may be useful in the diagnosis and treatment of gastric cancer.</p><p><b>METHODS</b>Data of expressed sequence tags (EST) and serial analysis of gene expression (SAGE) in Cancer Genome Anatomy Project (CGAP) were employed to analyze differential gene expression between normal and cancerous gastric epithelium,the obtained genes were further analyzed by virtual Northern blotting and compared with microarray data from Stanford Microarray Database (SMD).</p><p><b>RESULTS</b>NCBI digital differential display (DDD), cDNA digital gene expression displayed (DGED) and SAGE DGED produced 165,286 and 181 differential expression genes.All these genes were analyzed by virtual Northern blotting and 45 genes were obtained. Comparing with microarray data, candidate genes were reduced to 12. Further RT-PCR analyses validated 4 genes, including ANXA1, MSMB, ANXA10 and PSCA, were differentially expressed in normal and cancerous gastric tissues.</p><p><b>CONCLUSIONS</b>Combined multiple high throughput analysis and data mining is an effective strategy for identification of gastric cancer-related genes. Further analyses of these genes from data mining will provide biomarkers for the diagnosis and treatment of gastric cancer.</p>