title: Alantolactone induces cell apoptosis partially through down-regulation of testes-specific protease 50 expression.
abstract: Testes-specific protease 50 (TSP50) is aberrantly expressed in many cancer biopsies and plays a crucial role in tumorigenesis, which make it a potential cancer therapeutic target for drug discovery. Here, we constructed a firefly luciferase reporter driven by the TSP50 gene promoter to screen natural compounds capable of inhibiting the expression of TSP50. Then we identified alantolactone, a sesquiterpene lactone, could efficiently inhibit the promoter activity of TSP50 gene, further results revealed that alantolactone also efficiently inhibited the expression of TSP50 in both mRNA and protein levels. Moreover, we found alantolactone could increase the ratio of Bax/Bcl-2, and activate caspase-9 and caspase-3 in the cancer cells with high expression of TSP50, surprisingly, the same effects can also be observed in the same cells just by knockdown of TSP50 gene expression. Furthermore, our results suggested that overexpression of TSP50 decreased the cell sensitivity to alantolactone-induced apoptosis in those cancer cells. Taken together, these results suggest that alantolactone induces mitochondrial-dependent apoptosis at least partially via down-regulation of TSP50 expression.
authors.affiliation: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, 130024 Changchun, China; Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, 130024 Changchun, China.
authors.forename: Xu-Guang
authors.initials: XG
authors.lastname: Mi
authors.affiliation: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, 130024 Changchun, China.
authors.forename: Zhen-Bo
authors.initials: ZB
authors.lastname: Song
authors.affiliation: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, 130024 Changchun, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Wu
authors.affiliation: Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China.
authors.forename: Yu-Wei
authors.initials: YW
authors.lastname: Zhang
authors.affiliation: Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, 130024 Changchun, China.
authors.forename: Lu-Guo
authors.initials: LG
authors.lastname: Sun
authors.affiliation: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, 130024 Changchun, China. Electronic address: baoyl800@nenu.edu.cn.
authors.forename: Yong-Li
authors.initials: YL
authors.lastname: Bao
authors.affiliation: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, 130024 Changchun, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, 130024 Changchun, China.
authors.forename: Li-Hua
authors.initials: LH
authors.lastname: Zheng
authors.affiliation: Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, 130024 Changchun, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China.
authors.forename: Chun-Lei
authors.initials: CL
authors.lastname: Yu
authors.affiliation: Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Northeast Normal University, 130024 Changchun, China. Electronic address: wuy705@nenu.edu.cn.
authors.forename: Yin
authors.initials: Y
authors.lastname: Wu
authors.affiliation: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, 130024 Changchun, China.
authors.forename: Guan-Nan
authors.initials: GN
authors.lastname: Wang
authors.affiliation: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, 130024 Changchun, China; Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China.
authors.forename: Yu-Xin
authors.initials: YX
authors.lastname: Li
mesh_terms: D000935:Antifungal Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3 / Q000096:biosynthesis; D053453:Caspase 9 / Q000096:biosynthesis; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects / Q000648:ultrastructure; D015536:Down-Regulation / Q000187:drug effects; D006801:Humans; D007783:Lactones / Q000633:toxicity*; D008297:Male; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000187:drug effects* / Q000235:genetics; D045787:Sesquiterpenes, Eudesmane / Q000633:toxicity*; D014162:Transfection; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D007783:Lactones; D019253:Proto-Oncogene Proteins c-bcl-2; D045787:Sesquiterpenes, Eudesmane; D051028:bcl-2-Associated X Protein; D012313:RNA; D012697:Serine Endopeptidases; C446379:testis-specific protease 50; D053148:Caspase 3; D053453:Caspase 9; C004363:alantolactone
keywords: Alantolactone; Anticancer; Apoptosis; Drug screening; TSP50 inhibitor