title: Expression of hPOT1 in HeLa cells and the probability of gene variation of hpot1 Exon14 in endometrial cancer are much higher than in other cancers.
abstract: To investigate the expression of hPOT1 in the HeLa cell line and screen point mutations of hpot1 in different tumor tissues a two step osmotic method was used to extract nuclear proteins. EMSA was performed to determine the expression of hPOT1 in the HeLa cell line. PCR was also employed to amplify the exon14 sequence of the hpot1 gene in various of cancer tissues. A SV gel and PCR clean-up system was performed to enrich PCR products. DNAStar was used to analyse the exon14 sequence of the hpot1 gene. hPOT1 was expressed in the HeLa cell line and the signal was gradually enhanced as the amount of extracted nuclear proteins increased. The DNA fragment of exon14 of hpot1 was successfully amplified in the HeLa cell line and all cancer tissues, point mutations being observed in 2 out of 3 cases of endometrial cancer (66.7%) despite the hpot1 sequence being highly conserved. However, the sequence of hpot1 exon14 do not demonstrate point mutations in most cancer tissues. Since hPOT1 was expressed in HeLa cell and the probability of gene point variants was obviously higher in endometrial cancer than other cancers, it may be involved in the pathogenesis of gynecological cancers, especially in cervix and endometrium.
authors: Department of Clinical Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing, China.
authors: Fei
authors: F
authors: Liu
authors: Xiao-Yun
authors: XY
authors: Pu
authors: Shao-Guang
authors: SG
authors: Huang
authors: Gui-Ming
authors: GM
authors: Xiang
authors: Dong-Neng
authors: DN
authors: Jiang
authors: Gou
authors: G
authors: Hou
authors: Di-Nan
authors: DN
authors: Huang
mesh_terms: D000328:Adult; D000368:Aged; D001483:Base Sequence; D024202:Electrophoretic Mobility Shift Assay; D016889:Endometrial Neoplasms / Q000235:genetics*; D005091:Exons / Q000235:genetics*; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D009369:Neoplasms / Q000235:genetics*; D017354:Point Mutation / Q000235:genetics*; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000089804:Shelterin Complex; D015996:Survival Rate; D034501:Telomere-Binding Proteins / Q000235:genetics*
chemical_list: C440849:POT1 protein, human; D012333:RNA, Messenger; D000089804:Shelterin Complex; D034501:Telomere-Binding Proteins