Related Experiment Video

Updated: Apr 27, 2026

Intracellular Phosphoflow Cytometry of Acute Myeloid Leukemia Patient-Derived Xenotransplants
07:38

Intracellular Phosphoflow Cytometry of Acute Myeloid Leukemia Patient-Derived Xenotransplants

Published on: June 6, 2025

945

Expression of RKIP in chronic myelogenous leukemia K562 cell and inhibits cell proliferation by regulating the

Bin Li, Baolan Sun, Jianwei Zhu

    Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine
    |July 13, 2014
    PubMed

    Insights

    RAF kinase inhibitor protein (RKIP) regulates cell signaling. Overexpressing RKIP in chronic myelogenous leukemia (CML) cells reduced viability and proliferation while increasing apoptosis, offering potential therapeutic strategies.

    Area of Science:

    • Molecular Biology
    • Cancer Biology
    • Cell Signaling

    Background:

    • RAF kinase inhibitor protein (RKIP) is a known negative regulator of the RAS-MAPK/ERK signaling pathway.
    • Understanding RKIP's role in hematological malignancies like chronic myelogenous leukemia (CML) is crucial for developing targeted therapies.

    Purpose of the Study:

    • To investigate RKIP expression in CML K562 cells.
    • To determine the effects of RKIP modulation on CML cell characteristics and signaling pathways.

    Main Methods:

    • RKIP expression was modulated using pcDNA3.1-RKIP plasmid (overexpression) and RKIP-siRNA (knockdown) in K562 cells.
    • Cell viability was assessed by MTT assay, cell cycle and apoptosis by Flow Cytometry (FCM).
    • Clonogenic growth was evaluated using Colony Forming Unit (CFU) assays; signaling pathways (phospho-ERK1/2, NF-κB) were analyzed via Western blot and luciferase reporter assays.

    Main Results:

    • Transfection successfully altered RKIP expression, with overexpression significantly decreasing cell viability, proliferation, and clonogenic growth (p<0.05).
    • RKIP overexpression notably enhanced CML cell apoptosis (p<0.01).
    • Overexpression of RKIP strongly inhibited phospho-ERK1/2 levels and NF-κB transcriptional activity.

    Conclusions:

    • RKIP plays a significant inhibitory role in CML cell proliferation and survival.
    • Modulating RKIP levels impacts key signaling pathways (ERK, NF-κB) in CML.
    • RKIP represents a promising target for future biological therapies in myelogenous leukemia.

    More Related Videos

    Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle10:05

    Studying RNA Interactors of Protein Kinase RNA-Activated during the Mammalian Cell Cycle

    Published on: March 5, 2019

    6.1K
    Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation09:09

    Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation

    Published on: November 10, 2017

    7.9K

    Related Concept Videos

    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...
    7.3K
    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...
    4.8K
    Inhibition of CDK Activity02:34

    Inhibition of CDK Activity

    4.3K
    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...
    5.1K
    Interactions Between Signaling Pathways01:19

    Interactions Between Signaling Pathways

    Signaling cascades usually lack linearity. Multiple pathways interact and regulate one another, allowing cells to integrate and respond to diverse environmental stimuli.
    Convergence and divergence, and cross-talk between signaling pathways
    Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
    4.6K
    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...
    3.6K