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Silencing of IQGAP1 by shRNA inhibits the invasion of ovarian carcinoma HO-8910PM cells in vitro.

Dong PX, Jia N, Xu ZJ, Liu YT, Li DJ, Feng YJ - J. Exp. Clin. Cancer Res. (2008)

Bottom Line: Levels of both IQGAP1 mRNA and protein were significantly reduced in HO-8910PM cells transfected with plasmid-based IQGAP1-specific shRNAs.RNAi-mediated knockdown of IQGAP1 expression in HO-8910PM cells resulted in a significant decrease in cell invasion and migration.Our findings support the hypothesis that IQGAP1 promotes tumor progression and identify IQGAP1 as a potential therapeutic strategy for ovarian cancer and some other tumors with over-expression of the IQGAP1 gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Hospital and Institute of Obstetrics and Gynecology, Fudan University Shanghai Medical, College, Shanghai, PR China. dpx1cn@yahoo.com.cn

ABSTRACT

Background: IQGAP1 is a scaffolding protein and overexpressed in many human tumors, including ovarian cancer. However, the contribution of IQGAP1 to invasive properties of ovarian cancer cells remains unknown. Here, we investigated the effect of IQGAP1-specific short hairpin RNA (shRNA) expressing plasmids on metastatic potential of ovarian cancer HO-8910PM cells.

Methods: We used RT-PCR and Western blot analysis to characterize expression of IQGAP1 in three human ovarian cancer-derived cell lines SK-OV-3, HO-8910 and HO-8910PM. We then determined whether expression of endogenous IQGAP1 correlated with invasive and migratory ability by using an in vitro Matrigel assay and cell migration assay. We further knocked down IQGAP1 using shRNA expressing plasmids controlled by U1 promoter in HO-8910PM cells and examined the proliferation activity, invasive and migration potential of IQGAP1 shRNA transfectants using MTT assay, in vitro Matrigel-coated invasion assay and migration assay.

Results: IQGAP1 expression level seemed to be closely associated with the enhanced invasion and migration in ovarian cancer cell lines. Levels of both IQGAP1 mRNA and protein were significantly reduced in HO-8910PM cells transfected with plasmid-based IQGAP1-specific shRNAs. RNAi-mediated knockdown of IQGAP1 expression in HO-8910PM cells resulted in a significant decrease in cell invasion and migration.

Conclusion: Our findings support the hypothesis that IQGAP1 promotes tumor progression and identify IQGAP1 as a potential therapeutic strategy for ovarian cancer and some other tumors with over-expression of the IQGAP1 gene.

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indicates that IQGAP1 mRNA and protein levels in HO-8910PM-shIQGAP1, HO-8910PM-shRNA negative and un-transfected HO-8910PM cells were determined by RT-PCR and Western blot analysis, respectively.
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Figure 2: indicates that IQGAP1 mRNA and protein levels in HO-8910PM-shIQGAP1, HO-8910PM-shRNA negative and un-transfected HO-8910PM cells were determined by RT-PCR and Western blot analysis, respectively.

Mentions: To further investigate whether IQGAP-1 can induce cancer cell invasion and migration, we knocked down IQGAP1 using shRNA expressing plasmids controlled by U1 promoter (Clone 1, 2, 3 and 4) in HO-8910PM cells and selected by growth in the presence of puromycin. We first examined the knock-down efficiencies of different IQGAP1 shRNAs using Western blot analysis. HO-8910PM-shIQGAP1 cells (Clone 1, 2, 3 and 4) showed a significant decrease in IQGAP1 mRNA and protein expression when compared with HO-8910PM-shRNA negative cells or un-transfected HO-8910PM cells (Fig. 2). The above results demonstrated that the expression of IQGAP1 could be down-regulated specifically and effectively by specific IQGAP1 shRNA.


Silencing of IQGAP1 by shRNA inhibits the invasion of ovarian carcinoma HO-8910PM cells in vitro.

Dong PX, Jia N, Xu ZJ, Liu YT, Li DJ, Feng YJ - J. Exp. Clin. Cancer Res. (2008)

indicates that IQGAP1 mRNA and protein levels in HO-8910PM-shIQGAP1, HO-8910PM-shRNA negative and un-transfected HO-8910PM cells were determined by RT-PCR and Western blot analysis, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2626583&req=5

Figure 2: indicates that IQGAP1 mRNA and protein levels in HO-8910PM-shIQGAP1, HO-8910PM-shRNA negative and un-transfected HO-8910PM cells were determined by RT-PCR and Western blot analysis, respectively.
Mentions: To further investigate whether IQGAP-1 can induce cancer cell invasion and migration, we knocked down IQGAP1 using shRNA expressing plasmids controlled by U1 promoter (Clone 1, 2, 3 and 4) in HO-8910PM cells and selected by growth in the presence of puromycin. We first examined the knock-down efficiencies of different IQGAP1 shRNAs using Western blot analysis. HO-8910PM-shIQGAP1 cells (Clone 1, 2, 3 and 4) showed a significant decrease in IQGAP1 mRNA and protein expression when compared with HO-8910PM-shRNA negative cells or un-transfected HO-8910PM cells (Fig. 2). The above results demonstrated that the expression of IQGAP1 could be down-regulated specifically and effectively by specific IQGAP1 shRNA.

Bottom Line: Levels of both IQGAP1 mRNA and protein were significantly reduced in HO-8910PM cells transfected with plasmid-based IQGAP1-specific shRNAs.RNAi-mediated knockdown of IQGAP1 expression in HO-8910PM cells resulted in a significant decrease in cell invasion and migration.Our findings support the hypothesis that IQGAP1 promotes tumor progression and identify IQGAP1 as a potential therapeutic strategy for ovarian cancer and some other tumors with over-expression of the IQGAP1 gene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Hospital and Institute of Obstetrics and Gynecology, Fudan University Shanghai Medical, College, Shanghai, PR China. dpx1cn@yahoo.com.cn

ABSTRACT

Background: IQGAP1 is a scaffolding protein and overexpressed in many human tumors, including ovarian cancer. However, the contribution of IQGAP1 to invasive properties of ovarian cancer cells remains unknown. Here, we investigated the effect of IQGAP1-specific short hairpin RNA (shRNA) expressing plasmids on metastatic potential of ovarian cancer HO-8910PM cells.

Methods: We used RT-PCR and Western blot analysis to characterize expression of IQGAP1 in three human ovarian cancer-derived cell lines SK-OV-3, HO-8910 and HO-8910PM. We then determined whether expression of endogenous IQGAP1 correlated with invasive and migratory ability by using an in vitro Matrigel assay and cell migration assay. We further knocked down IQGAP1 using shRNA expressing plasmids controlled by U1 promoter in HO-8910PM cells and examined the proliferation activity, invasive and migration potential of IQGAP1 shRNA transfectants using MTT assay, in vitro Matrigel-coated invasion assay and migration assay.

Results: IQGAP1 expression level seemed to be closely associated with the enhanced invasion and migration in ovarian cancer cell lines. Levels of both IQGAP1 mRNA and protein were significantly reduced in HO-8910PM cells transfected with plasmid-based IQGAP1-specific shRNAs. RNAi-mediated knockdown of IQGAP1 expression in HO-8910PM cells resulted in a significant decrease in cell invasion and migration.

Conclusion: Our findings support the hypothesis that IQGAP1 promotes tumor progression and identify IQGAP1 as a potential therapeutic strategy for ovarian cancer and some other tumors with over-expression of the IQGAP1 gene.

Show MeSH
Related in: MedlinePlus