Screening imprinted genes during malignant transformation of immortalized human osteoblastic cells by large-scale oligomicroarray technique
- VernacularTitle:高密度Oligo基因芯片筛选人永生化成骨细胞恶性转化中印迹基因的研究
- Author:
Gang MENG
;
Yi LI
;
Qiaonan GUO
- Publication Type:Journal Article
- Keywords:
osteosarcoma;
imprinted gene;
DNA microarray
- From:Journal of Third Military Medical University
2003;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the pathogenesis-related imprinted genes in osteosarcoma and investigate the role of imprinted genes expression in osteosarcoma pathogenesis.Methods The model of malignant transformed immortalized human osteoblastic cells was established and the malignant transformed cells were collected on day 40,55,70,90.The untreated cells passaged from the normal cells on day 90 served as control.After the total RNA extraction,the finally synthesized biotinylated cDNAs were hybridized to CapitalBio Genechip CAP-F0017.2 of the differentially expressed genes.Two genes were chosen at random to further confirm the array results using the SYBR Green real-time PCR in samples of control,day 40 and day 90. Results By an entrance limit of ≥2.0 or ≤0.5,ten imprinted genes which biological functions are mainly related with cell growth/maintenance and signaling transduction were detectable for notablely differential expression,including down-regulated genes of CD81,GRB10,NDN,MEST and up-regulated genes of H19,MKRN3,SLC22A1L,TSSC3,CDKN1C in the malignant transformation of immortalized human osteoblastic cells.The array results were further confirmed by the real-time RT-PCR.Conclusion Ten imprinted genes were detectable for notablely differential expression.Our results will promote our understanding of the molecular mechanisms of imprinted genes in the course of osteosarcoma pathogenesis.
