[Apoptosis of HL-60 cells induced by recombinant common Buckwheat trypsin inhibitor].
The study was purposed to investigate the apoptosis of HL-60 cells induced by recombinant common buckwheat trypsin inhibitor (rBTI) and its mechanism. The inhibition rate of rBTI on HL-60 cells was detected by MTT (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide); the morphology of HL-60 nuclei was observed by fluorescence microscopy; the apoptosis cells of HL-60 detected by agarose gel electrophoresis and the changes of apoptosis rate was assayed by flow cytometry (FCM), when the HL-60 cells were treated with different concentration of rBTI for 24 hours. The results showed that the growth of HL-60 cells was inhibited evidently after treatment with rBTI in a dose-dependent manner, but there were minimal effects on normal human peripheral blood mononuclear cells (PBMNCs). The nuclei of HL-60 cells showed the characteristics of apoptosis, the analysis by flow cytometry indicated that the apoptosis rate of HL-60 cells was 52% after treatment with rBTI (100 microg/ml), DNA analyzed by agarose gel electrophoresis showed "ladder" pattern. It is concluded that rBTI obviously inhibits growth of HL-60 and induces its apoptosis which provides a foundation for use of recombinant common buckwheat trypsin inhibitor to cure the acute myeloid leukemia.
Gao
Li
L
Key Laboratory for Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China.
Li
Yu-Ying
YY
Zhang
Zheng
Z
Wang
Zhuan-Hua
ZH
Wang
Hong-Wei
HW
Zhang
Li
L
Zhu
Lei
L
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D019612:Fagopyrum / Q000737:chemistry*; D018922:HL-60 Cells; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D014361:Trypsin Inhibitors / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*
D000970:Antineoplastic Agents; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors