Inhibition of ovarian cancer growth by small interfering RNA targeting X-linked inhibitor of apoptosis gene
- VernacularTitle:X染色体相关凋亡抑制蛋白的小分子干扰RNA对卵巢上皮性癌细胞生长的影响
- Author:
Jia-Jia MA
;
Bi-Liang CHEN
;
Xiao-Yan XIN
;
- Publication Type:Journal Article
- Keywords:
Ovarian neoplasms;
RNA;
small interfering;
X-linked inhiritor of apoptosis
- From:
Chinese Journal of Obstetrics and Gynecology
2001;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct an RNA interference vector to down-regulate X-linked inhibitor of apoptosis(XIAP)gene and study the RNA interference effect on the cell cycle and growth of ovarian cancer.Methods Oligonucleotides of 64 base pairs for hairpin RNA targeting XIAP were designed, chemically synthesized,annealed,and cloned into the pSUPER vector.After identification by restriction digestion,the correct vectors were transiently transfected into SKOV3 cells,a human ovarian cancer cell line.The XIAP mRNA was detected by RT-PCR.The proteins were detected by western blot and indirect immunofluorescence staining.Flow cytometry(FCM)analysis and methyl thiazolyl tetrazolium(MTT)assay method were applied to measure cell cycle,cell growth and sensitiveness to cisplatin.Results SKOV3 cells had a high level expression of XIAP.The vector of RNA interference,which can interfere with XIAP gene was successfully constructed.After transient transfection,the expression of XIAP protein was significantly decreased in SKOV3 cells and the value of relative density was 3584?124,2138?65,1973?80 and 110 ?12,respectively(P=0.0334).At the same time,the expression of XIAP mRNA was decreased accordingly and the value of relative density was 6674?274,4532?107,2322?57 and 1864?78, respectively(P=0.0127).The FCM results showed that,the vector could increase the number of cells in G_1 phase compared with parent cells and compared with the cells transfected with pSUPER(P
