Centrosome is the major microtubule organizing center in mammalian cells. The centrosome plays key roles in the formation of the mitotic spindle, cell polarity and cell locomotion. In a typical somatic cell, the centrosome is composed of a pair of centrioles that are surrounded by a mass of amorphous pericentriolar material(PCM). The centrosomal-associated protein, hNinein, has been identified as a microtubules minus end capping, centrioles position, centrosome maturation and anchoring protein. Recently data reveal ninein CCII domain is associated with centrosomal targeting signal, regulating signal and phosphorylation sites, suggesting that this protein could contribute to centrosomal targeting signal as well as regulate asymmetry centrosomes. In this report, we therefore examine whether four C-terminal of hNinein splicing isoforms, including isoform 1, isoform 2, isoform 5 and a newly finding isoform 6, represent differential characterization, expression and functional aspects. To investigate the expression level of hNinein isoform 6 in variant cancer cell lines and astrocytoma patients. Isoform 6 only exist in IMR32 cancer cell line ( neuroblastoma cell line) and 42¢Mastrocytoma patients. We speculate that isoform 6 might play a role and exist in nerve cell or neuron. Using immunofluorescence assay, we show that all three hNinein isoforms are concentrated at centrosomes in HeLa cells, but not isoform 6. Overexpression of isoform 6 in HeLa cell might influence the distribution of