title: [Inducing-apoptosis effect of brucine on human monocytic leukemia cell line THP-1 and its mechanism].
abstract: This study was aimed to investigate the inducing-apoptosis effect of brucine on human monocytic leukemia cell line THP-1 cells and its possible mechanism. The inhibition effect of brucine on growth of THP-1 cells was measured by CCK-8 method. Morphological changes of THP-1 cells treated with brucine was detected by acridine orange/ethidium bromide (AO/EB)double staining. Annexin-V/PI double labeling method was used to assay the apoptosis rate of THP-1 cells. The effect of brucine on THP-1 cell cycle distribution was detected by PI single staining. RT-PCR was used to detect the expression of BCL-2 and BAX. The results showed that the brucine could inhibit the THP-1 cell growth in concentration and time-dependent manners at the range of 50 to 400 µg/ml. The cells stained with AO/EB revealed that the brucine induced the nuclear chromatin condensation. After the THP-1 cells were treated with brucine of 400µg/ml for 48 hours, most nucleic were stained as orange-red, and condensed, displaying the late apoptotic cell morphology. Annexin-V/PI detection showed that brucine could induce apoptosis of THP-1 cells in a concentration-dependent manner. Compared with the control group, more cells in brucine-treated group were arrested at G0/G1 phase in a concentration-dependent manner. RT-PCR detection revealed that the expression of BCL-2 was down-regulated strikingly and BAX was up-regulated. It is concluded that brucine can efficiently inhibit cell growth and block THP-1 cells in G0/G1 phase. The mechanism of THP-1 cell apoptosis induced by brucine may be related to the inhibition of BCL-2 and activation of BAX.
authors.affiliation: First clinicial Department, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Xin
authors.affiliation: Department of Hematology, Peace Hospital Affiliated to Changzhi Medical college, Changzhi 046000, Shanxi Province, China. E-mail: weiwu88@sohu.com.
authors.forename: Wu
authors.initials: W
authors.lastname: Wei
authors.affiliation: Department of Hematology, Peace Hospital Affiliated to Changzhi Medical college, Changzhi 046000, Shanxi Province, China.
authors.forename: Ai-Fang
authors.initials: AF
authors.lastname: Ji
authors.affiliation: Department of Hematology, Peace Hospital Affiliated to Changzhi Medical college, Changzhi 046000, Shanxi Province, China.
authors.forename: Xu-Liang
authors.initials: XL
authors.lastname: Shen
authors.affiliation: Department of Hematology, Peace Hospital Affiliated to Changzhi Medical college, Changzhi 046000, Shanxi Province, China.
authors.forename: Guo-Xiang
authors.initials: GX
authors.lastname: Zhang
authors.affiliation: Department of Hematology, Peace Hospital Affiliated to Changzhi Medical college, Changzhi 046000, Shanxi Province, China.
authors.forename: Mei-Xiang
authors.initials: MX
authors.lastname: Zhang
authors.affiliation: First clinicial Department, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China.
authors.forename: Xian-Xian
authors.initials: XX
authors.lastname: Li
authors.affiliation: First clinicial Department, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China.
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Zhang
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013331:Strychnine / Q000031:analogs & derivatives* / Q000494:pharmacology; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: C494826:BAX protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C083806:brucine; D013331:Strychnine