Retrovirus-mediated RNA inhibition of EphA2 gene expression in colon adenocarcinoma HCT-8 cells / 中华病理学杂志
Chinese Journal of Pathology
;
(12): 101-105, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-258213
ABSTRACT
<p><b>OBJECTIVE</b>To explore novel cancer gene therapy by retrovirus-mediated RNAi technique to suppress the endogenous EphA2 oncogene expression in colon adenocarcinoma HCT-8 cells.</p><p><b>METHODS</b>Sequence information of EphA2 mRNA was selected and two complementary oligonucleotides with hairpin loop were designed. Retrovirus-mediated RNAi expression vector (pSIREN-EphA2) was then constructed and transfected into the HCT-8 cells. Inhibition of EphA2 protein expression was quantitatively determined by Western blot and immunohistochemistry assay (SP method).</p><p><b>RESULTS</b>The construction of pSIREN-EphA2 vector was successful and confirmed by restriction enzyme analysis and DNA sequencing. The post-transfection level of EphA2 protein expressions was greatly reduced in HCT-8 cells transfected with pSIREN-EphA2, as compared with those of untransfected cells and the vector control (P < 0.001).</p><p><b>CONCLUSIONS</b>EphA2 protein expression in HCT-8 cell line can be suppressed using recombinant retrovirus-mediated RNAi technique. This approach may provide a novel gene therapy against colonic adenocarcinoma.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pathology
/
Retroviridae
/
Recombinant Proteins
/
RNA, Messenger
/
Transfection
/
Adenocarcinoma
/
Down-Regulation
/
Gene Expression Regulation, Neoplastic
/
Colonic Neoplasms
/
Reverse Transcriptase Polymerase Chain Reaction
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Pathology
Year:
2006
Type:
Article
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