Related Experiment Video

Updated: Feb 16, 2026

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
10:06

Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells

Published on: April 26, 2017

9.5K

Aberrant alternative splicing of RHOA is associated with loss of its expression and activity in diffuse-type gastric

Shingo Miyamoto1, Yuko Nagamura1, Ayaka Nakabo2

  • 1Department of Cancer Cell Research, Sasaki Institute, Sasaki Foundation, 2-2 Kandasurugadai, Chiyoda-ku, Tokyo 101-0062, Japan.

Biochemical and Biophysical Research Communications
|December 17, 2017
PubMed

Insights

Aberrant alternative splicing of RhoA leads to its loss of activity and expression in diffuse-type gastric carcinoma (DGC) cells, as identified in this study. This finding highlights a novel mechanism contributing to DGC development.

Area of Science:

  • Molecular Biology
  • Oncology
  • Genetics

Background:

  • RhoA, a Rho family small GTPase, regulates crucial cellular functions.
  • Recurrent somatic mutations in RHOA have been identified in diffuse-type gastric carcinoma (DGC), implicating it as a driver gene.
  • Gastric carcinoma (GC) is a significant global health concern, necessitating research into its underlying molecular mechanisms.

Purpose of the Study:

  • To investigate potential abnormalities in RhoA expression and activity within a panel of gastric carcinoma (GC) cell lines.
  • To elucidate the molecular mechanisms behind RhoA dysregulation in DGC.
  • To determine the functional consequences of RHOA alterations in gastric cancer.

Main Methods:

  • Pulldown assays and immunoblot analysis to assess RhoA activity and expression.
  • RHOA coding region sequencing to identify mutations and splicing abnormalities.
  • Quantitative real-time PCR (qRT-PCR) to analyze gene expression levels of wild-type RHOA and its splicing variants.

Main Results:

  • RhoA activity and expression were detected in most GC cell lines, but absent in two DGC cell lines (HSC-59 and GSU).
  • Aberrant alternative splicing of RHOA was identified in these DGC cell lines, with near-undetectable wild-type RHOA and exclusive expression of splicing variants.
  • Low expression levels of RHOA splicing variants prevented protein detection, and ectopic expression of these isoforms did not restore protein expression or activity.

Conclusions:

  • Aberrant alternative splicing of RHOA is a key mechanism leading to the loss of its functional activity and expression in DGC.
  • This RHOA dysregulation contributes to the pathogenesis of diffuse-type gastric carcinoma.
  • Targeting RHOA splicing or its downstream pathways may offer therapeutic strategies for DGC.
Keywords:
Alternative splicingDiffuse-typeGastric carcinomaPulldown assayRho family small GTPaseRhoA

More Related Videos

Detection of Alternative Splicing During Epithelial-Mesenchymal Transition11:48

Detection of Alternative Splicing During Epithelial-Mesenchymal Transition

Published on: October 9, 2014

13.4K
RhoC GTPase Activation Assay09:58

RhoC GTPase Activation Assay

Published on: August 22, 2010

13.1K

Related Concept Videos

Alternative RNA Splicing02:18

Alternative RNA Splicing

Alternative RNA splicing is the regulated splicing of exons and introns to produce different mature mRNAs from a single pre-mRNA. Unlike in constitutive splicing where a single gene produces a single type of mRNA, alternative splicing allows an organism to produce multiple proteins from a single gene and plays an important role in protein diversity.
There are five types of alternative RNA splicing that vary in the ways the pre-mRNA segments are removed or retained in the mature mRNA. The first...
25.3K
Alternative RNA Splicing02:18

Alternative RNA Splicing

5.2K
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...
7.4K
RNA Splicing01:32

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
60.8K
Small GTPases - Ras and Rho01:24

Small GTPases - Ras and Rho

Ras and Rho are small monomeric GTPases that act downstream of receptor tyrosine kinase (RTK) and regulate various cellular processes. These GTPases switch between active and inactive states by binding to guanine nucleotides.
Three regulatory proteins control their activity:
5.5K
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...
5.3K