Purpose: This study was designed to investigate the role of homeodomain-interacting protein kinase 2(HIPK2) in the apoptosis of T lymphocytes which was mediated by the deprivation of cytokine. Methods: CTLL-2, interleukin-2(IL-2)-dependent T lymphocytes, was deprived of IL-2 to induce apoptosis. Apoptotic cells were quantified by the measurement of the hypodiploid area in the PI-staining profiles. Apoptosis of CTLL-2 was also determined by DNA fragmentation analysis after agarose gel electrophoresis. The expression of genes involved in apoptosis was detected by real-time quantitative RT-PCR. Antisense technology was applied to suppress the expression of HIPK2 in order to investigate the role of HIPK2 in the apoptosis of CTLL-2 mediated by the deprivation of IL-2. Results: The percentage of apoptotic CTLL-2 increased over time with IL-2 deprivation, which resulted in detectable DNA ladder. Analysis of the gene expression involved in the apoptosis of CTLL-2 revealed that there was no change found for Fas or FasL (P > 0.05). However, the expression of Bim was increased (P < 0.01) and the expression of Bcl-x was decreased (P < 0.05). The up-regulation of HIPK2 was also found (P < 0.05). After the expression of HIPK2 was down-regulated, the percentage of apoptotic CTLL-2 decreased consequently (P < 0.01). In addition, the expression of Bim decreased (P < 0.01) and that of Bcl-x increased (P < 0.05). Conclusions: HIPK2 promoted the apoptosis of CTLL-2 deprived of IL-2, which was associated with the increased expression of Bim and the decreased expression of Bcl-x.