title: The role of c2orf68 and PI3K/Akt/mTOR pathway in human colorectal cancer.
abstract: The aim of the research is to determine whether c2orf68 gene plays a role in the carcinogenesis of human colorectal cancer and to study the function of c2orf68 belonging to the UPF0561 family. The mRNA expression levels of c2orf68 were examined in 30 pairs of human colorectal adenocarcinoma tissues and adjacent normal colorectal tissues by qRT-PCR. The SW480 and SW620 cell lines were transfected with siRNA against the c2orf68 gene and set gene. The expressed mRNA levels of Akt, PI3K, Bax, Bcl-2, caspase3, c-Myc, cyclinD1, pp2a and set were determined by qRT-PCR, and the protein levels of C2ORF68, c-Myc, PP2A and SET were examined by Western blot. Cell proliferation was tested by MTT assay, and apoptosis and cell cycle were studied by flow cytometry. Cancer metastasis assay was performed by transwell chamber. The c2orf68 mRNA expression was down-regulated in 63.33 % of the cancer samples, and a positive correlation was found between the mRNA expression of c-Myc and pp2a that of c2orf68. Meanwhile, there was a negative correlation between the mRNA expression of c2orf68 and set. The c2orf68 mRNA was significantly down-regulated in SW480(-c2orf68) and SW620(-c2orf68) cells. The inhibitory rate in the two cell lines was, respectively, 65.2 and 71.6 % by qRT-PCR. A 22.7 % inhibition on cell proliferation in SW480(-c2orf68) cells and a 21.2 % inhibition in SW620(-c2orf68) cells were observed using the MTT assay. Flow cytometry analysis indicates that the cell apoptosis rate was 21.42 % in SW480(-c2orf68) cells and 17.78 % in SW620(-c2orf68) cells, whereas the percentage of G1 phase cells was 61.8 and 58.6 % in SW480(-c2orf68) and SW620(-c2orf68) cells, respectively. In addition, the mRNA expression of set and Bax was up-regulated after c2orf68 interfered in SW480(-c2orf68) and SW620(-c2orf68) cells, whereas that of Bcl-2, c-Myc, cyclinD1, caspase3 and pp2a was down-regulated. Consistent with the mRNA results, the protein expression of C2ORF68, PP2A and c-Myc was down-regulated, whereas that of SET was up-regulated. Our data thus suggest that c2orf68 promotes carcinogenesis through the regulation of mammalian target of rapamycin signaling pathway.
authors.affiliation: Department of Anatomy, College of Basic and Forensic Medicine, Sichuan University, Chengdu, Sichuan, People's Republic of China.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Wen
authors.forename: Jianjun
authors.initials: J
authors.lastname: Zhu
authors.forename: Lei
authors.initials: L
authors.lastname: Dong
authors.forename: Yao
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000328:Adult; D017209:Apoptosis / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D004268:DNA-Binding Proteins; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D016259:Genes, myc; D056488:Histone Chaperones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering; D015398:Signal Transduction / Q000235:genetics; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D056488:Histone Chaperones; D009363:Neoplasm Proteins; D019869:Phosphatidylinositol 3-Kinases; D054648:Protein Phosphatase 2; D051057:Proto-Oncogene Proteins c-akt; D034741:RNA, Small Interfering; D058570:TOR Serine-Threonine Kinases; D014157:Transcription Factors; C000597591:C2ORF68 protein, human; C513473:SET protein, human; C546842:MTOR protein, human