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@# Objective: To screen related genes of melanoma-associated antigen-A11 (MAGE-A11) in breast cancer cells based on highthroughput DNAmicroarray technology, and to validate from the aspects of quantity and function. Methods: DNAmicroarray was used to screen the differently-expresseddown-stream mRNAs of MAGE-A11 in breast cancercelllines (MCF-7, MDA-MB-231 and BT-549). Cluster analysis was applied on representative genes and quantitative RT-PCR was used to validate. CCK-8, scratch wound healing assay and Transwell assaywere used to detect the effect of MAGE-A11 on the proliferation,migration and invasion of breast cancer cells. Results: Over-expression of MAGE-A11 caused the differential expression of 1608 down-stream genes in 3 breast cancer cell lines, which was associated with various cell functions such as protein ubiquitination,cell proliferation and apoptosis, tumor invasion and metastasis.qRT-PCR validated that the expression of ZNF-451, CENPTJ, CDK13, API5 and LMO7, which were highly expressed in microarray, were also significantly higher than those in control group (P<0.01);in addition, SHPRH, PML, MARK2, LIMA1 and ANGPTL4, which were low-expressed in microarray, were also significantly lower than those in control group (P<0.01). MAGE-A11transfection directly increased the proliferation of breast cancer MCF-7, MDA-MB-231 and BT-549 cells at 72 h (all P<0.01); compared with control group after transfectionexhibited obvious wound healing at 48 h (P<0.05 or P<0.01) and significantly increased trans-membrane cell numbers (all P<0.01). Conclusion: Many differentially expressed genes related to ubiquitination, cell proliferation and apoptosis, tumor invasion and migration were screened in MCF-7, MDA-MB-231 and BT-549 breast cancer cells. Among them, 10 typical differentially expressed genes were identified in terms of quantity and function.
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Objective To study the effect of rhubarb extract on the gene expression profile in diabetic nephropathy rats, and analyze the mechanism of rhubarb on diabetic nephropathy in gene expression level. Methods Male rats without any treatments were used as the normal control group. Diabetic nephropathy was induced by streptozotocin in the rats as the diabetic nephropathy group. The diabetic nephropathy rats were treated with rhubarb extract as the rhubarb-treated group and with placebo as negative control group. The renal cortex was obtained from four groups. Cy5 or Cy3 fluorescent labeled DNA probes were prepared from mRNA and hybridized against the BioDoor Genechip, finally fluorescent signals were assayed and raw data was analyzed. Results There were about 8. 4 % genes altered in gene expression profile. The expression level of 12 genes were downregulated while 323 genes were upregulated significantly in the renal cortex of the diabetic animals. Compared with the negative control rats, 120 genes were downregulated and eight genes were upregulated significantly in the rhubarb-treated group, which occupied 3.3% of the total genes on the genechip. Furthermore, the expression changes of partial altered genes in the diabetic nephropathy group were reversed by rhubarb treatment. Conclusion Rhubarb is effective on the downregulation or upregulation of some differential expression genes, and can partially reverse streptozotocin-induced gene expression profile.