[Study on NB4 cell apoptosis induced by trichosanthin]
Wen-Da Luo1, Chang-Ming Ren, Min Zhu
1Department of Hematology, Taizhou Hospital of Zhejiang Province, Linhai 317000, China.
Abstract:
In order to study the influence of trichosanthin (TCS) on apoptosis and growth inhibition of human NB4 cells in vitro, the expression of annexin V and the change of DeltaPsim of NB4 cells induced by TCS was analyzed by FACS, and MTT assay was adopted to measure the growth inhibition ratio of NB4 cells treated with TCS. Apoptosis was assayed by agarose gel electrophoresis. The results showed the higher concentration of TCS and the longer the acting time, the stronger growth inhibition of NB4 cells. The expression of annexin V was positive, and the positive ratio was greatly enhanced with prolongation of acting time. DeltaPsim reduced gradually while the apoptosis cells increasing. DNA agarose gel electrophoresis showed a gradient, which confirmed that TCS could induce NB4 cells apoptosis. In conclusion, taken together, data show that TCS can inhibit NB4 growth in vitro, and induce apoptosis. Experiment provides an important evidence for application of TCS in clinical treatment of acute promyelocytic leukemia.
Insights
Trichosanthin (TCS) effectively inhibits human NB4 cell growth and induces apoptosis in vitro. This study provides evidence for TCS
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Context:
- Acute promyelocytic leukemia (APL) is a hematologic malignancy.
- Developing novel therapeutic agents for APL is crucial.
- Trichosanthin (TCS), a protein from Trichosanthes kirilowii, has shown cytotoxic properties.
Purpose:
- To investigate the effects of TCS on the apoptosis and growth inhibition of human NB4 cells in vitro.
- To analyze the expression of annexin V and mitochondrial membrane potential (DeltaPsim) in TCS-treated NB4 cells.
- To confirm apoptosis induction by TCS using DNA agarose gel electrophoresis.
Summary:
- MTT assays demonstrated that higher concentrations and longer exposure times of TCS resulted in stronger growth inhibition of NB4 cells.
- Flow cytometry analysis revealed increased annexin V expression and decreased DeltaPsim, indicating apoptosis.
- DNA agarose gel electrophoresis confirmed dose-dependent DNA fragmentation, characteristic of apoptosis.
Impact:
- Data indicate that TCS inhibits NB4 cell proliferation and induces apoptosis in vitro.
- This research provides evidence supporting the potential clinical application of TCS in treating acute promyelocytic leukemia.
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