|Public on Nov 25, 2013
|clear renal cell carcinoma cell line 786-O, stable transfection with knockdown of S100A6
|cell line: clear cell renal cell carcinoma cell line 786-O
genotype/variation: knockdown of S100A6
|The 786-O cells were stable transfection with overexpression and knockdown of S100A6, each treatment had a vector transfection control. The final purity of stable transfection is 99%.
|The 786-O cells were cultured in RPMI 1640 medium (HyClone), and the culture medium was supplemented with 10% fatal bovin serum (FBS, Gibco). The cells were maintained at 37℃ with 5% CO2.
|Total RNA from each sample was extracted using TRK1001（LC Sciences, Cat. TRK-1001, Total RNA Purification Kit）according to the manufacturer’s instructions. Total RNA was quantified by the NanoDrop ND-2000 (Thermo Scientific) and the RNA integrity was assessed using Agilent Bioanalyzer 2100 (Agilent Technologies).
|The sample labeling, microarray hybridization and washing were performed based on the manufacturer’s standard protocols. Briefly, total RNA were transcribed to double strand cDNA, then synthesized into cRNA and labeled with Cyanine-3-CTP.
|The labeled cRNAs were hybridized onto the microarray according to the manufacturer’s standard protocols.
|After washing, the arrays were scanned by the Agilent Scanner G2505C (Agilent Technologies).
|Sample name: SH1
Total 1399 different expression genes (DEGs) including 880 up-regulated and 519 down regulated genes were found in overexpression S100A6 and vector control group.
|Feature Extraction software (version10.7.1.1, Agilent Technologies) was used to analyze array images to get raw data. Genesrping were employed to finish the basic analysis with the raw data. To begin with, the raw data was normalized with the quantile algorithm. The probes that at least 1 out of all samples have flags in Detected were chosen for further data analysis. Differentially expressed genes were then identified through fold change as well as P value calculated with t-test. The threshold set for up- and down-regulated genes was a fold change>= 2.0 and a P value<= 0.05. expression pattern among samples.
|Nov 25, 2013
|Last update date
|Apr 23, 2018
|PLA general hospital
|The mechanism of elevated S100A6 (Calcyclin) Enhances Tumorigenesis in clear cell renal cell carcinoma