Construction of let-7d expression vector and its inhibitory effect on HMGA2 and ras expression in human ovarian cancer cells in vitro / 南方医科大学学报
Journal of Southern Medical University
;
(12): 1752-1757, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-352341
ABSTRACT
<p><b>OBJECTIVE</b>To elucidate the role of let-7d in regulating the biological behavior of ovarian cancer cells and their expressions of HMGA2 and ras proteins.</p><p><b>METHODS</b>The pre-let-7d sequence was synthesized and inserted into pcDNA6.2GW/EmGFPmiR and transfected into ovarian cancer IGROV1 cells to cause pre-let-7d overexpression. Real-time quantitative RT-PCR was employed to examine the expression levels of let-7d miRNA and HMGA2 mRNA, and Western blotting was performed to detect the expressions of HMGA2 and ras protein in the transfected cells. The effect of pcDNA6.2GW-let-7d transfection on IGROV1 cell proliferation was determined using MTT assay and the cell apoptosis rate was measured using flow cytometry.</p><p><b>RESULTS</b>The eukaryotic expression vector containing the target gene let-7d was successfully constructed and transfected into IGROV1 cells. The transfected cells showed a marked reduction of HMGA2 expression but a less obvious down-regulation of ras expression. Transfection with pcDNA6.2GW-let-7d to suppress the expression of HMGA2 caused alterations of the phenotype of IGROV1 cells shown by a reduced proliferative activity and increased cell apoptosis.</p><p><b>CONCLUSION</b>Let-7d plays an important role in altering the malignant cell phenotype of ovarian cancer IGROV1 cells by regulating the expression of HMGA2.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Ovarian Neoplasms
/
Pathology
/
Plasmids
/
Gene Expression Regulation, Neoplastic
/
Ras Proteins
/
HMGA2 Protein
/
MicroRNAs
/
Cell Line, Tumor
/
Genetics
/
Metabolism
Limits:
Female
/
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2012
Type:
Article
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