To investigate the in situ expression of the genes in large numbers of cultured cells, a high-throughput technique was established. A novel method of constructing the cell microarrays was devised and the cell microarrays including 100 spot arrays of mixture of the graft cells and paraffin from 20 kinds of cell lines was made successfully. Expression of P53, P21, PTEN and P16 protein in the cell microarrays was detected by immunohistochemistry. Meanwhile, expression of BRD7 and NGX6 mRNA in the cell microarrays was estimated by in situ hybridization. In situ expression profiles of P53, P21, PTEN, P16 protein and BRD7, NGX6 mRNA in different kinds of cultured cells were established. The cell microarrays provided a new high-throughput tool for biological function studies of the genes. The cell microarrays could be widely used the in situ analysis of the gene expression at the DNA, RNA, and protein level. In additional, the cell microarrays could also be applied to estimate drug target validations.