Related Experiment Video

Updated: Aug 9, 2026

Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment
10:59

Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment

Published on: November 23, 2017

RNA interference as a key to knockdown overexpressed cyclooxygenase-2 gene in tumour cells

A Strillacci1, C Griffoni, E Spisni

  • 1Department of Experimental Biology, University of Bologna, and Center for Applied Biomedical Research (CRBA), St Orsola-Malpighi University Hospital, Bologna, Italy.

British Journal of Cancer
|April 20, 2006
PubMed

Insights

This study demonstrates that RNA interference targeting cyclooxygenase-2 (COX-2) effectively reduces colon cancer cell growth. A retroviral vector enhanced this gene silencing, offering a potential targeted cancer therapy with fewer side effects.

Area of Science:

  • Oncology
  • Molecular Biology
  • Gene Therapy

Background:

  • Overexpression of Cyclooxygenase-2 (COX-2) is common in various cancers, particularly colon cancer, contributing to tumor progression.
  • Current COX-2 inhibitor treatments face concerns due to potential long-term side effects.
  • RNA interference (RNAi) presents a novel strategy for gene silencing in cancer research.

Purpose of the Study:

  • To investigate the efficacy of RNA interference (RNAi) in downregulating COX-2 gene expression in colon cancer cells.
  • To evaluate the potential of a retroviral vector system for enhanced COX-2 gene silencing.
  • To assess the impact of COX-2 knockdown on the malignant behavior of colon cancer cells in vitro.

Main Methods:

  • Designing and utilizing specific short hairpin RNA (shRNA) sequences targeting COX-2 mRNA.
  • Employing a retroviral vector (pSUPER.retro) for efficient delivery of anti-COX-2 shRNA into HT29 colon cancer cells.
  • Assessing gene silencing potency, interferon response, and phenotypic changes in COX-2 deficient cells.

Main Results:

  • Sequences targeting COX-2 mRNA effectively downregulated gene expression and inhibited angiogenesis in vitro.
  • The retroviral vector significantly enhanced COX-2 silencing in HT29 cells without inducing an interferon response.
  • COX-2 deficient HT29 cells exhibited impaired in vitro malignant behavior.
  • Higher infection efficiency was observed in tumor cells compared to normal endothelial cells.

Conclusions:

  • Retroviral delivery of RNAi targeting COX-2 is an effective tool for understanding COX-2's role in colon cancer.
  • This approach offers a promising strategy for targeted cancer therapy by specifically silencing COX-2 in tumor cells.
  • The method allows for specific tumor cell treatment while potentially preserving normal endothelial COX-2 activity.

More Related Videos

DNA Vector-based RNA Interference to Study Gene Function in Cancer13:10

DNA Vector-based RNA Interference to Study Gene Function in Cancer

Published on: June 4, 2012

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells07:23

Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells

Published on: May 30, 2025

Related Concept Videos

Experimental RNAi02:15

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
Abnormal Proliferation02:23

Abnormal Proliferation

Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the daughter...
Cancer-Critical Genes II: Tumor Suppressor Genes01:05

Cancer-Critical Genes II: Tumor Suppressor Genes

Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
Cancer-Critical Genes II: Tumor Suppressor Genes01:05

Cancer-Critical Genes II: Tumor Suppressor Genes

Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
Inhibition of Cdk Activity02:34

Inhibition of Cdk Activity

The orderly progression of the cell cycle depends on the activation of Cdk protein by binding to its cyclin partner. However, the cell cycle must be restricted when undergoing abnormal changes. Most cancers correlate to the deregulated cell cycle, and since Cdks are a central component of the cell cycle, Cdk inhibitors are extensively studied to develop anticancer agents. For instance, cyclin D associates with several Cdks, such as Cdk 4/6, to form an active complex. The cyclin D-Cdk4/6 complex...
Cancer-Critical Genes I: Proto-oncogenes01:33

Cancer-Critical Genes I: Proto-oncogenes

Genes usually encode proteins necessary for the proper functioning of a healthy cell. Mutations can often cause changes to the gene expression pattern, thereby altering the phenotype.
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...