title: Multisite mutation of monomer survivin with enhanced effect on apoptosis regulation of breast cancer cells.
abstract: Survivin is an important protein in regulating both cell apoptosis and proliferation. It has attracted growing attentions in recent years as a promising target for cancer therapy. Previous studies have revealed that monomeric survivin regulated apoptosis in a more significant way than the wild-type survivin that generally contains a large portion of its dimers. In order to investigate the roles of monomeric mutant survivin apoptosis and cell cycle regulation of human cancer cells, we developed and tested three dominant-negative mutants with multisite mutations (MSM) including TAT-survivin(34/101/102), TAT-survivin(34/117/101/102) and TAT-survivin(117/101/102). Results revealed that MSM mutants remained as monomers under ambient conditions, and induced cells (breast cancer Bcap-37 cells) apoptosis even more efficiently, primarily through the caspase-dependent and Bcl-2-related pathways, than non-monomeric mutants. We further identified that the TAT-survivin(34/101/102) and TAT-survivin(117/101/102) MSM significantly inhibited the proliferation of Bcap-37 cells and arrested cells in S and G2/M phases, while TAT-survivin(34/117/101/102) arrested cells in G2/M phase. It appeared to us that TAT-survivin(34/101/102) and TAT-survivin(117/101/102) also inhibited cell proliferation more significantly. These findings suggest that such MSM afford monomeric survivin with promising potentials for cancer therapy.
authors: State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, East China University of Science and Technology, Shanghai 200237, China; Department of Bioproducts and Biosystems Engineering and Biotechnology Institute, University of Minnesota, St. Paul, MN 55108, USA.
authors: Guoying
authors: G
authors: Dai
authors: Department of Bioproducts and Biosystems Engineering and Biotechnology Institute, University of Minnesota, St. Paul, MN 55108, USA; Shanghai Key Laboratory of New Drug Design, School of pharmacy, East China University of Science and Technology, Shanghai 200237, China.
authors: Wenyun
authors: W
authors: Zheng
authors: State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, East China University of Science and Technology, Shanghai 200237, China. Electronic address: maxy@ecust.edu.cn.
authors: Xingyuan
authors: X
authors: Ma
authors: State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, East China University of Science and Technology, Shanghai 200237, China; Department of Bioproducts and Biosystems Engineering and Biotechnology Institute, University of Minnesota, St. Paul, MN 55108, USA. Electronic address: ping@umn.edu.
authors: Ping
authors: P
authors: Wang
mesh_terms: D017209:Apoptosis; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D002453:Cell Cycle; D045744:Cell Line, Tumor; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D000077022:Survivin
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D050505:Mutant Proteins; D000077022:Survivin
keywords: Apoptosis; Breast cancer; Cell cycle regulation; Protein engineering; Site-specific mutagenesis; Survivin