[Expression of RhoC in gastric cancer cell lines and construction and identification of RhoC-specific siRNA expression vector].
OBJECTIVE
To study the expression of RhoC in human gastric cancer cell lines and to construct and identify the RhoC-specific siRNA expressing vector.


METHODS
The expression of RhoC in human gastric cancer cell lines was detected by Western blot. According to the computer aided design(CAD),RhoC-specific siRNA gene was synthesized and cloned into the expression vector mU6pro. The constructed RhoC-siRNA was transiently transfected into high-metastatic gastric cancer cell line AGS and the inhibition effect of RhoC-siRNA on expression of RhoC in AGS cells was detected using Western blot.


RESULTS
The expression level of RhoC was up-regulated in high metastatic cells lines. Double enzyme digestion analysis and DNA sequencing confirmed that the RhoC-specific siRNA expression vector was constructed successfully. RhoC expression in AGS cells was specifically suppressed after transfection of RhoC-siRNA.


CONCLUSIONS
The RhoC-specific siRNA expression vector has been successfully constructed, which may provide a novel applicable strategy for gene therapy of gastric cancer.
Liu
Na
N
Institute of Digestive Disease, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China. liuzhp@edu.cn
Bi
Feng
F
Pan
Yang-lin
YL
Xue
Yan
Y
Han
Zhe-yi
ZY
Liu
Chang-jiang
CJ
Fan
Dai-ming
DM
D045744:Cell Line, Tumor; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D009362:Neoplasm Metastasis; D034741:RNA, Small Interfering / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013274:Stomach Neoplasms / Q000378:metabolism* / Q000473:pathology; D014162:Transfection; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000075024:rhoC GTP-Binding Protein
D034741:RNA, Small Interfering; C066690:RHOC protein, human; D020741:rho GTP-Binding Proteins; D000075024:rhoC GTP-Binding Protein