| Objective:To investigate the Proliferation inhibition and apoptosis induction effect of hirudin on human nasopharyngeal cancer CNE2 cells and the possible mechanisms.Methods:The human nasopharyngeal carcinoma cell line CNE2 was assigned to different concentration. Inhibitory effect of cell proliferation was measured by MTT assay. And calculate the IC50 at different time periods. The cell apoptosis was detected by flow Cytometry(FCM) after annexin-V and PI dual labeling. The cell cycle of CNE2 cells was detected FCM. Quantitative real-time polymerase chain reaction was used to demonstrate expression levels of involved RNA such as p21, Bax.Results:Apparently, Hirudin could inhibit CNE2 cells proliferation, with the increasing of concentration and time of hirudin on CNE2 cells (p<0.05). The IC50 at 24h、48h and 72h was(8.28±2.1) ATU/ml、(5.11±0.7) ATU/ml and (4.23 ±0.5)ATU/ml respectively. The apoptosis rate of the group treated by hirudin raised in evidence (p<0.05), moreover, with the same concentration of hirudin at the longer time, the apoptosis rate was raised (p<0.05). For hirudin treated group, the arresting of the cell cycle was in G2/M phase. According to the results of the expression levels of involved RNA, p21 and Bax were significantly upregulated in cells treated with hirudin.Conclusion:In this study, the apoptosis induced effect of hirudin on human nasopharyngeal carcinoma CNE2 cell line was analyzed. The hirudin can inhibit growth, induce CNE2 cell G2/M cycle arrest, also can induce apoptosis of CNE2 cells. The findings presented here reveal that the G2/M cycle arrest treated with hirudin was associated with the upregulation of p21, and Bax was also raised when treated with hirudin, which would trigger apoptosis. These data provide preclinical support for chemotherapeutic approach with hirudin for the treatment of nasopharyngeal cancer. All these results indicate that hirudin may represent a novel class of chemotherapeutic agents for nasopharyngeal cancer. |
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