Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Jul 1;23(7):2387-2395.
doi: 10.31557/APJCP.2022.23.7.2387.

Novel Approach Using shRNA of IQGAP1 for Colon Cancer Therapy: HCT116 as a Surrogate Model Colorectal Carcinoma

Khairy M M A Zoheir  1 , Khaled Mahmoud  2 , Gamal I Harisa  3   4 , Abdelkader Ashour  5 , Hala E Abdel-Hamied  6 , Amro A Amara  7 , Karima F Mahrous  1 , Ahmed A Abd-Rabou  8
Affiliations

Novel Approach Using shRNA of IQGAP1 for Colon Cancer Therapy: HCT116 as a Surrogate Model Colorectal Carcinoma

Khairy M M A Zoheir et al. Asian Pac J Cancer Prev. .

Abstract

Background: Colorectal carcinoma (CRC) represents life-threatening problems worldwide. IQ motif containing GTPase activating protein 1 (IQGAP1) is acting as oncogenesis regulators. RNAi is proposed as promising cancer therapeutics.

Objective: The objective of this work to explore the consequences of the IQGAP1 silence as a goal for treating CRC using the HCT166 cells as a model for human colon cancer.

Methods: RNAi technology was used to design a short specific sequence of RNA (shRNA) to silence the IQGAP1 oncogene. The impact of IQGAP1 silencing on IQGAPs, Ras, IL-8, and TRAIL was investigated. Furthermore, the effect of IQGAP1 silencing on cell viability, proliferation, apoptosis, and invasive capacity was investigated.

Results: The present results revealed that IQGAP1 shRNA-treated HCT166 cells showed no invasive capacity compared to the control cells. The silencing of IQGAP1 induced remarkable downregulation of IQGAP1, RAS (H&K), IL-8, CXCR1, CXCR2, NF-kB, BCL-2, and apoptosis of HCT166 cells. On the contrary, IQGAP2, IQGAP3, DR4, DR5, CASP-3, and BAX genes were significantly up-regulated.

Conclusion: The IQGAP1 regulates the expression of IQGAPs, Ras, IL-8 receptors, and the apoptotic network. Therefore, the silence of IQGAP1 is a promising strategy for colon cancer therapy.

Keywords: Apoptosis; Cell migration; Colon cancer; IQGAP1; shRNA.

PubMed Disclaimer

Conflict of interest statement

The authors declared no potential conflicts of interest

Figures

Figure 1
Figure 1
Showing Cell Proliferation by MTT Assay. HCT166 cells were incubated with 5μM of IQGAP1 shRNA vector for 24 48, 72 and 72 h
Figure 2
Figure 2
Healthy Population of HCT166 Cells was Counted Using Flow Cytometry up on Treatment with 5μM of IQGAP1 shRNA Vector for 24 and 48 h. The experiment was repeated 3 independent times (n=3).
Figure 3
Figure 3
Wound Healing Assay of IQGAP1siRNA on HCT166 Cells. (After 48 h)
Figure 4
Figure 4
Expressions of mRNA for Definite Genes after HCT166 Cell Treatment with IQGAP1 shRNA Vector. (a-f) * indicated significantly differences between means at P <0.05 and error bars represents standard error of mean (SEM). Means comparisons were performed by using Duncan’s multiple range test
Figure 5
Figure 5
Protein Expression of IQGAP1, KRAS, BAX, casp-9 and cleaved casp-3 Using Western Blot Analysis
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Bashour AM, Fullerton AT, Hart MJ, Bloom GS. IQGAP1, a Rac- and Cdc42-binding protein, directly binds and cross-links microfilaments. J Cell Biol. 1997;137:1555–1566. - PMC - PubMed
    1. Chen F, Zhu HH, Zhou LF, Wu SS, Wang J, Chen Z. IQGAP1 is overexpressed in hepatocellular carcinoma and promotes cell proliferation by Akt activation. Exp Mol Med. 2010;42:477–483. - PMC - PubMed
    1. Chen WQ, Zheng RS, Zhang SW, Li N, Zhao P, Li GL, Wu LY, He J. Report of incidence and mortality in china cancer registries, 2008. Chin J Cancer Res. 2012;24:171–80. - PMC - PubMed
    1. Colin D, Hema K, Dmitri V, Zhigang L, Robert D, David B. Valentina A IQGAP1 and IQGAP2 are reciprocally altered in hepatocellular carcinoma. BMC Gastroenterol. 2010;10:125. - PMC - PubMed
    1. Dong P, Nabeshima K, Nishimura N. Overexpression and diffuse expression pattern of IQGAP1 at invasion fronts are independent prognostic parameters in ovarian carcinomas. Cancer Lett. 2006;243:120–12. - PubMed

MeSH terms

Substances