[Growth inhibitory effects of lipofectamine-mediated DCC gene on ovarian epithelial carcinoma].
OBJECTIVE
To study the inhibitory effects of lipofectamine-mediated deleted colorectal carcinoma gene on ovarian epithelial carcinoma (ovarian cancer) cell line SKOV3.


METHODS
We constructed a recombinant eukaryotic expression vector pcDNA3.1 (+)-DCC containing exogenous human DCC cDNA and vector with neomycin resistance gene, which were introduced by lipofectamine-mediated gene transfection into SKOV3 cell line that does not express DCC endogenously, thus forming SKOV3/DCC. Therefore, the experimental cells were classified into SKOV3/DCC, SKOV3/Neo and SKOV3. By using reverse transcriptase-polymerase chain reaction and immunocytochemistry, the expression of DCC mRNA and its protein were examined.


RESULTS
Exogenous DCC had successfully been transferred into SKOV3 cells and obtained permanent expression. The growth speed of SKOV3/DCC was slower than the other two groups, there was significant difference between them (P < 0.01). SKOV3/DCC clones number was 38 +/- 8, while SKOV3 and SKOV3/Neo were 192 +/- 8 and 186 +/- 10, respectively, there was significant difference between them (P < 0.01). The percentage of G(1) phase cells increased to 78.0%, which that of S phase decreased to 5.3% by analyzing cell cycle, there was significant difference between them (20.0% and 3.2%, P < 0.01). The ultrastructural changes of the cells were observed under electron microscope, revealing growth retardation.


CONCLUSIONS
DCC gene played an important role in generation and development of ovarian carcinomas.
Li
Pei-ling
PL
Department of Obstetrics and Gynecology, Second Affiliated Hospital of Harbin Medical University, Harbin 150086, China.
Hu
Chun-jie
CJ
Li
Chang-min
CM
Meng
Chun-yan
CY
Gao
Lei
L
D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D000075389:DCC Receptor; D005260:Female; D005822:Genetic Vectors / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008055:Lipids / Q000737:chemistry*; D010051:Ovarian Neoplasms / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods; D025521:Tumor Suppressor Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology
D000075389:DCC Receptor; C501909:DCC protein, human; D008055:Lipids; C086724:Lipofectamine; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D025521:Tumor Suppressor Proteins