title: Notch3 overexpression causes arrest of cell cycle progression by inducing Cdh1 expression in human breast cancer cells.
abstract: Uncontrolled cell proliferation, genomic instability and cancer are closely related to the abnormal activation of the cell cycle. Therefore, blocking the cell cycle of cancer cells has become one of the key goals for treating malignancies. Unfortunately, the factors affecting cell cycle progression remain largely unknown. In this study, we have explored the effects of Notch3 on the cell cycle in breast cancer cell lines by 3 methods: overexpressing the intra-cellular domain of Notch3 (N3ICD), knocking-down Notch3 by RNA interference, and using X-ray radiation exposure. The results revealed that overexpression of Notch3 arrested the cell cycle at the G0/G1 phase, and inhibited the proliferation and colony-formation rate in the breast cancer cell line, MDA-MB-231. Furthermore, overexpressing N3ICD upregulated Cdh1 expression and resulted in p27(Kip) accumulation by accelerating Skp2 degradation. Conversely, silencing of Notch3 in the breast cancer cell line, MCF-7, caused a decrease in expression levels of Cdh1 and p27(Kip) at both the protein and mRNA levels, while the expression of Skp2 only increased at the protein level. Correspondingly, there was an increase in the percentage of cells in the G0/G1 phase and an elevated proliferative ability and colony-formation rate, which may be caused by alterations of the Cdh1/Skp2/p27 axis. These results were also supported by exposing MDA-MB-231 cells or MCF-7 treated with siN3 to X-irradiation at various doses. Overall, our data showed that overexpression of N3ICD upregulated the expression of Cdh1 and caused p27(Kip) accumulation by accelerating Skp2 degradation, which in turn led to cell cycle arrest at the G0/G1 phase, in the context of proliferating breast cancer cell lines. These findings help to illuminate the precision therapy targeted to cell cycle progression, required for cancer treatment.
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.|b Dept. of Pathology, and The Breast Center , Cancer Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Chun-Fa
authors.initials: CF
authors.lastname: Chen
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Dou
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.|b Dept. of Pathology, and The Breast Center , Cancer Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Yuan-Ke
authors.initials: YK
authors.lastname: Liang
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.|b Dept. of Pathology, and The Breast Center , Cancer Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Hao-Yu
authors.initials: HY
authors.lastname: Lin
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.|b Dept. of Pathology, and The Breast Center , Cancer Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Jing-Wen
authors.initials: JW
authors.lastname: Bai
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.|b Dept. of Pathology, and The Breast Center , Cancer Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Xi-Xun
authors.initials: XX
authors.lastname: Zhang
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.|b Dept. of Pathology, and The Breast Center , Cancer Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Xiao-Long
authors.initials: XL
authors.lastname: Wei
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Yao-Chen
authors.initials: YC
authors.lastname: Li
authors.affiliation: a Department of Thyroid and Breast Surgery, Changjiang Scholar's Laboratory , The First Affiliated Hospital of Shantou University Medical College , Shantou , China.|b Dept. of Pathology, and The Breast Center , Cancer Hospital of Shantou University Medical College , Shantou , China.
authors.forename: Guo-Jun
authors.initials: GJ
authors.lastname: Zhang
mesh_terms: D015703:Antigens, CD; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology; D015820:Cadherins / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000528:radiation effects; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics / Q000378:metabolism; D005260:Female; D059585:G1 Phase Cell Cycle Checkpoints; D006801:Humans; D061986:MCF-7 Cells; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D011839:Radiation, Ionizing; D060888:Real-Time Polymerase Chain Reaction; D000071656:Receptor, Notch3 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D044786:S-Phase Kinase-Associated Proteins / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000528:radiation effects
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; C499374:NOTCH3 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000071656:Receptor, Notch3; D044786:S-Phase Kinase-Associated Proteins; D050760:Cyclin-Dependent Kinase Inhibitor p27
keywords: Cdh1; Notch3; Skp2; breast cancer; cell cycle; p27Kip