ShRNA-mediated gene silencing of beta-catenin inhibits growth of human colon cancer cell Colo205 in vitro / 中华胃肠外科杂志
Chinese Journal of Gastrointestinal Surgery
;
(12): 301-305, 2009.
Article
in Chinese
| WPRIM
| ID: wpr-326508
ABSTRACT
<p><b>OBJECTIVE</b>To observe the gene silencing and disruption of WNT pathway mediated by the specific shRNA targeted against beta-catenin and its effect on cell proliferation of the human colon cancer cell line Colo205.</p><p><b>METHODS</b>The shRNA plasmid vectors against beta-catenin were constructed and transfected into Colo205 cells with Lipofectamine 2000. The expression of beta-catenin was detected by RT-PCR and Western blot. Immunofluorescence staining was also performed to detect the beta-catenin protein expression in cells. The cell proliferation inhibition was determined by MTT assay and soft agar colony formation assay.</p><p><b>RESULTS</b>The shRNA vectors targeted against beta-catenin were successfully constructed and efficiently suppressed the expression of beta-catenin mRNA and protein(P<0.05). The expression inhibition rates were 47.89% and 45.26% at the mRNA and protein level respectively. Immunofluorescence microscopy also demonstrated the inhibition of beta-catenin protein induced by these specific shRNAs. The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates in time-dependent manner. At 72 h post-transfection, the cell viability of CAT group was 48.5%, which was significantly different as compared with that of blank control group's 91.3%(P<0.05). In the soft agar, there were 9, 46, 43 cell colonies in the CAT, blank, and negative control groups respectively, which were significantly different(P<0.05).</p><p><b>CONCLUSIONS</b>The specific shRNAs targeted against beta-catenin has a gene silencing effect and blocks the WNT signaling pathway, which can inhibit the growth of Colo205 cells.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
In Vitro Techniques
/
Transfection
/
Signal Transduction
/
Colonic Neoplasms
/
RNA Interference
/
Cell Line, Tumor
/
Cell Proliferation
/
Wnt Proteins
/
Beta Catenin
/
Genetics
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Gastrointestinal Surgery
Year:
2009
Type:
Article
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