Abstract:
Objective To screen the genes which are related to stress response closely in the malignant proliferation of hepatocellular carcinoma cells SMMC-7721 against different stressors in vitro.
Methods We conventionally cultured SMMC-7721 cells in vitro, grouping the cells in log phase into normal group, G418 group and calcium group. Each treatment group was given the corresponding drug respectively. After 72h and 4h treatment, we collected the cells and extracted total RNA. The GeneAtlas microarray system was used to make Affymetrix Human Gene 1.1 ST Array Strip genechip. iterPlier was used for calculating gene expression, to work out preliminary statistical processing. We observed the characteristics of genes expression related to stress response in hepatocellular carcinoma cells. Then, qRT-PCR was used to detect the relative expression of each gene.
Results Eventually we screened 53 genes which had a large quantity of expression and the change were consistent with those under the effect of G418 and calcium chloride, moreover, 17 genes had relatively consistent trend with genechips verified by qRT-PCR.
Conclusion In the process of hepatocellular carcinoma cells SMMC-7721 resisting G418 and calcium chloride stress, 17 genes are important and verified relatively consistent trend by genechips and qRT-PCR.