1.Implication of expression of Nanog in prostate cancer cells and their stem cells.
Chen, GONG ; Hui, LIAO ; Fengjin GUO ; Liang, QIN ; Jun, QI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):242-6
Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.
2.Differential regulation of SREBP 1 c/cm on transcriptional activity and expression of PERK promoter
Qin HU ; Yu MAO ; Jialing LU ; Weiwei XIE ; Wei ZHENG ; Fengjin GUO
Basic & Clinical Medicine 2017;37(2):162-168
Objective To investigate the effect of sterol regulating element binding protein (SREBP1c) and its ac-tive form (SREBP1cm) on human protein kinase R-like endoplasmic reticulum kinase (PERK).Methods Re-porter victors of PERK promoter and its truncations were constructed with pGL 3-basic and co-transfected with internal reference pRL-SV40 into cell and luciferase activity was detected .pcDNA3.1 ( -)-SREBP1c or pcDNA3.1 ( -)-SREBP1cm was co-transfected with PERK promoter transcriptional activity core regions and the detection of dual -lu-ciferase reporter gene was used to analyze the regulation of SREBP 1c/1cm on PERK promoter transcriptional activity . The expression level of PERK mRNA and protein were detected by RT-PCR and Western blot .Results PERK pro-moter and truncations were successfully constructed into pGL 3-basic, and PERK promoter core area of transcription-al activity had determined;Dual-luciferase report gene showed that SREBP 1c inhibited PERK promoter transcrip-tional activity and SREBP1cm promoted PERK promoter transcriptional activity .RT-PCR and Western blot showed that SREBP1c decreased PERK mRNA and protein expression , but SREBP1cm increased PERK mRNA and protein expression, which was consistent with the detection of dual-luciferase report gene .Conclusions SREBP1c and SREBP1cm have a opposite regulation effect on PERK promoter transcriptional activity and its expression .
3.Cytogenetic and molecular characterization of a patient with partial 6q trisomy and 1q monosomy.
Fengjin QIN ; Xiaoyan LU ; Yapei FENG ; Peihong TANG ; Gang NIU ; Fan LI ; Jianhai ZHANG
Chinese Journal of Medical Genetics 2016;33(2):231-234
OBJECTIVETo analyze a girl with moderate mental retardation and speech and language disorders with cytogenetics technique and next-generation sequencing (NGS).
METHODSG-banding chromosome analysis was used to ascertain the karyotype of the child and her parents, and NGS was used for determining the size and origin of the abnormal chromosome fragment. Mate-pair and PCR were used to determine its parental origin.
RESULTSThe karyotype of the child was determined to be 46,XX,add(1)(q44)dn, while her parents were both normal. NGS revealed that the child has harbored a partial trisomy of 6q24.3-q27, and the breakpoint was mapped to at 6q24.3q27. In addition, a 2.5 Mb microdeletion at 1q44 was found in the patient.
CONCLUSIONNo recognizable phenotype was associated with 1q44 deletion. The abnormal phenotypes presented by the child may be attributed to the 6q24.3-q27 triplication. Compared with conventional cytogenetic analysis, NGS has a much higher resolution and great accuracy.
Adult ; Child ; Chromosome Banding ; Chromosome Disorders ; genetics ; Chromosomes, Human, Pair 1 ; genetics ; Chromosomes, Human, Pair 6 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Intellectual Disability ; genetics ; Male ; Monosomy ; genetics ; Trisomy ; genetics
4.Implication of Expression of Nanog in Prostate Cancer Cells and Their Stem Cells
GONG CHEN ; LIAO HUI ; GUO FENGJIN ; QIN LIANG ; QI JUN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(2):242-246
Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells.The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells,and fhe relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells.By using magnetic active cell sorting (MACS),we isolated a population of CD44+/CD133+ prostate cancer cells that display stem cell characteristics from PC3 cell line.lmmunohistochemistry revealed positive expressions of CD44,C D 133 and α2β1-integin in the isolated cells.CCK-8 analysis showed that isolated cells had a strong proliferafive ability.The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing,indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line.Western blotting exhibited that the isolated cells had higher experession of Nanog,an embryonic stem marker,as compared with PC3 cells.Our study showed that Nanog might be helpful in sustaining the self-renewal and fhe undifferentiation of prostate cancer stem cells,and may serve as a marker for prostate cancer stem cells for isolation and identification.