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The Study of Survivin in the Mechanisms of the Apoptosis Induced by Daunorubicin in Leukemic Cells

Author: HouYan
Tutor: HuQun
School: Huazhong University of Science and Technology
Course: Pediatrics
Keywords: Daunorubicin Survivin Apoptosis Leukemia
CLC: R733.7
Type: Master's thesis
Year: 2006
Downloads: 33
Quote: 0
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Abstract


By detecting the changes of different chemotherapy drugs after survivin and caspase-3, 8, 9, to explore the role of survivin DNR induced leukemia cell apoptosis mechanism. Different concentration of daunorubicin (DNR) and vincristine (VCR) (0.1 mg / L, 1 mg / L, 10 mg / L) were acting on Molt-4 cells at different time points: ( 1) MTT assay cell proliferation inhibition rate; (2) flow cytometry (FCM) analysis of the rate of apoptosis; (3) reverse transcription - polymerase chain reaction (RT-PCR) to detect changes in survivin mRNA content; (4) immunohistochemical method to detect survivin protein expression changes; (5) immunoblot (Western blot) detection of caspase-3, 8, 9 protein expression. Immunohistochemical method detected 62 cases of acute leukemia in children survivin protein expression, positioning, analysis of the correlation of survivin and P53, and the survivin and chemosensitivity relationship will. The results of the first part: (1) MTT results showed that with the increase in drug dose and longer duration of action, reduced inhibition of cell proliferation, in a time-and dose-dependent, and the the DNR role than VCR obvious (P lt; 0.05). (2) FCM analysis showed that in the same dose and the same time, DNR group compared VCR group of Molt-4 cell apoptosis rate was significantly higher (P lt; 0.01). (3) RT-PCR assay found in the DNR group, increased with dose and longer duration of action, reduced survivin mRNA contents of (P lt; 0.05), while no significant change (P gt the VCR group; 0.05). (4) immunohistochemical method can be seen to increase with dose and longer duration of action, DNR survivin protein expression reduce (P lt; 0.01) presents the phenomenon of transfer from the cytoplasm to the nucleus, and no significant changes in the VCR group (P gt; 0.05). (5) Western blot method observed the DNR group Molt-4 cells can be detected expression of caspase-3 zymogen activation fragments, while the expression of caspase-9 zymogen shear activation fragments detected in a time-and dose-dependent ( P lt; 0.05), but no significant expression of caspase-8 activation fragment. The second part: (1) Survivin expression in acute leukemia cells than normal cells (P lt; 0.05). (2) children with acute leukemia survivin expression was positively correlated (P lt; 0.01) and P53. (3) Survivin nuclear expression of acute leukemia in children than the cytoplasm by clinical complete remission rate (P lt; 0.05). Conclusion (1) DNR-induced apoptosis in leukemia cells through mitochondrial pathway. (2) Survivin/P53 play an important role in apoptosis induced by DNR, no significant relationship with VCR-induced apoptosis. (3) Survivn high in children with acute leukemia the expression prompted to play an important role in the development of acute leukemia (AL), survivin in the nucleus and cytoplasm of different expression as a prognostic indicator of tumor suppressor gene P53 inactivation of survivin expression in the AL in a synergistic effect.

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CLC: > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Leukemia
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