title: CSN5 silencing inhibits invasion and arrests cell cycle progression in human colorectal cancer SW480 and LS174T cells in vitro.
abstract: CSN5 has been implicated as a candidate oncogene in human cancers by genetic linkage with activation of the poor-prognosis, wound response gene expression signature. The present study aimed to investigate the effect of silencing CSN5 on invasion and cell cycle progression of human colorectal cancer cells, and to determine the potential molecular mechanisms that are involved. The CSN5 specific small interfering RNA (shRNA) plasmid vector was constructed and then transfected into colorectal cancer cells. The expression of CSN5 mRNA and protein was detected by quantitative polymerase chain reaction and western blot analysis, respectively. Cell adhesion and invasion were analyzed using MTS and Transwell assays, respectively, and cell cycle progression was analyzed using flow cytometry. Adhesion, invasion, and cell cycle distribution were assessed following knockdown of CSN5 by RNA interference (RNAi). Furthermore, knockdown of CSN5 significantly inhibited cell adhesion and reduced the number of invasive cells, while increasing the percentage of cells in the G0/G1 phase (P<0.05). Western blot and real-time PCR analysis were used to identify differentially expressed invasion and cell cycle associated proteins in cells with silenced CSN5. The expression levels of CSN5 in colorectal cancer cells transfected with siRNA were decreased, leading to a significant inhibition of colorectal cancer cell adhesion and invasion. Western blot analysis revealed that silencing of CSN5 may inhibit CD44, matrix metalloproteinase (MMP) 2 and MMP 9 protein expression, significantly promoted cell cycle-related genes P53 and P27 expression. In addition, CSN5 silencing may induce activation PI3K/AKT signal regulated cell invasion. Moreover, CSN5 silencing inhibited the secretion of TGF-β, IL-1β and IL-6 and the transcriptional activity of transcription factor NF-κB and Twist in human colorectal cancer cells. Taken together, down regulation of CSN5 may inhibit invasion and arrests cell cycle progression in colorectal cancer via PI3K/AKT/NF-κB signal pathway, which indicates that there is a potential of targeting CSN5 as a novel gene therapy approach for the treatment of colorectal cancer.
authors.affiliation: Department of Gastrointestinal Surgery, Tianjin Nankai Hospital Tianjin 300100, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Zhong
authors.affiliation: Department of Abdominal Tumor Surgery, Tianjin Medical University Cancer Institute Hospital Tianjin 300060, China.
authors.forename: Huikai
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Gastrointestinal Surgery, Tianjin Nankai Hospital Tianjin 300100, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Shan
authors.affiliation: Department of Gastrointestinal Surgery, Tianjin Nankai Hospital Tianjin 300100, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Zhang
mesh_terms: D000075686:COP9 Signalosome Complex; D002448:Cell Adhesion; D059447:Cell Cycle Checkpoints; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D016207:Cytokines / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D009361:Neoplasm Invasiveness; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection
chemical_list: D018797:Cell Cycle Proteins; D016207:Cytokines; D047908:Intracellular Signaling Peptides and Proteins; D016328:NF-kappa B; D012333:RNA, Messenger; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; D010447:Peptide Hydrolases; C501488:COPS5 protein, human; D000075686:COP9 Signalosome Complex
keywords: CSN5; colorectal cancer; invasion