Related Experiment Video

Updated: Aug 11, 2026

An In Vitro Dormancy Model of Estrogen-sensitive Breast Cancer in the Bone Marrow: A Tool for Molecular Mechanism Studies and Hypothesis Generation
08:48

An In Vitro Dormancy Model of Estrogen-sensitive Breast Cancer in the Bone Marrow: A Tool for Molecular Mechanism Studies and Hypothesis Generation

Published on: June 30, 2015

Gene expression profile by inhibiting Raf-1 protein kinase in breast cancer cells

Rajshree R Mewani1, Song Tian, Bihua Li

  • 1Department of Radiation Medicine, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC 20057, USA.

International Journal of Molecular Medicine
|February 9, 2006
PubMed

Insights

Inhibiting Raf-1 kinase in breast cancer cells with antisense oligonucleotides reduced tumor cell growth and identified 17 downstream genes involved in critical cellular processes like apoptosis and metabolism.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Genomics

Background:

  • Raf-1 protein serine-threonine kinase is crucial for cell growth, proliferation, and survival.
  • Antisense raf oligonucleotide (AS-raf-ODN)-mediated inhibition of Raf-1 has shown potential in arresting tumor growth and enhancing radiosensitization and chemosensitization.
  • Raf-1 inhibition is linked to apoptotic cell death, suggesting its role in regulating programmed cell death pathways.

Purpose of the Study:

  • To identify downstream target genes regulated by Raf-1 kinase using microarray gene expression analysis.
  • To investigate the impact of Raf-1 inhibition on gene expression profiles in breast cancer cells.
  • To gain insights into Raf-1-related signaling pathways and discover potential therapeutic targets.

Main Methods:

  • Treatment of MDA-MB-231 breast cancer cells with AS-raf-ODN.
  • Microarray gene expression analysis to compare gene profiles between treated and control cells.
  • Validation of microarray findings using Real-time PCR, Northern analysis, and Western analysis.

Main Results:

  • AS-raf-ODN treatment significantly inhibited Raf-1 protein and c-raf-1 mRNA levels in breast cancer cells.
  • A total of 17 genes, including c-raf-1, were identified with altered expression (4 upregulated, 13 downregulated) following AS-raf-ODN treatment.
  • Functional clustering revealed affected genes involved in apoptosis, cell adhesion, metabolism, signal transduction, and transcriptional regulation.

Conclusions:

  • Raf-1 inhibition impacts a network of genes involved in fundamental cellular processes.
  • The study provides a comprehensive view of Raf-1's downstream targets and signaling pathways.
  • This research establishes a model system for identifying potential therapeutic target genes in cancer treatment.

More Related Videos

Exploring the Pharmacological Action and Molecular Mechanism of Salidroside in Inhibiting MCF-7 Cell Proliferation and Migration11:13

Exploring the Pharmacological Action and Molecular Mechanism of Salidroside in Inhibiting MCF-7 Cell Proliferation and Migration

Published on: June 9, 2023

Related Concept Videos

mTOR Signaling and Cancer Progression03:03

mTOR Signaling and Cancer Progression

The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
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...
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a rapamycin-insensitive companion...
Targeted Cancer Therapies02:57

Targeted Cancer Therapies

The targeted cancer therapies, also known as “molecular targeted therapies,” take advantage of the molecular and genetic differences between the cancer cells and the normal cells. It needs a thorough understanding of the cancer cells to develop drugs that can target specific molecular aspects that drive the growth, progression, and spread of cancer cells without affecting the growth and survival of other normal cells in the body.
There are several types of targeted therapies against specific...
The Ras Gene02:38

The Ras Gene

The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
Ras is a superfamily...