Related Experiment Video

Updated: Apr 28, 2026

Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells
07:19

Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

Published on: September 28, 2011

38.0K

Identification of miRNAs that specifically target tumor suppressive KLF6-FL rather than oncogenic KLF6-SV1 isoform

Wei-Cheng Liang1, Yan Wang2, Li-Jia Xiao3

  • 1Croucher Laboratory for Human Genomics, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, P.R. China; School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, P.R. China.

RNA Biology
|June 13, 2014
PubMed

Insights

Specific microRNAs target the tumor-suppressive KLF6-FL isoform, promoting liver cancer. This discovery reveals a new mechanism where microRNAs drive tumorigenesis by inhibiting KLF6-FL, not the oncogenic KLF6-SV1.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Gene Regulation

Background:

  • The Krüppel like factor 6 (KLF6) gene produces multiple isoforms via alternative mRNA splicing, with implications in hepatocarcinogenesis.
  • KLF6 full-length (KLF6-FL) acts as a tumor suppressor, while KLF6 splice variant 1 (KLF6-SV1) is oncogenic, exhibiting opposing functions in liver cancer.
  • MicroRNAs (miRNAs) are small non-coding RNAs crucial for regulating gene expression by targeting specific mRNA regions.

Purpose of the Study:

  • To identify specific miRNAs that target the tumor-suppressive KLF6-FL isoform.
  • To investigate the functional consequences of miRNA targeting on KLF6-FL and KLF6-SV1.
  • To explore the role of identified miRNAs in liver cancer development and their potential epigenetic regulation.

Main Methods:

  • Bioinformatics analysis to predict miRNA targets.
  • Luciferase reporter assays to validate miRNA-binding specificity.
  • In vitro experiments to assess the functional effects of KLF6-FL and miR-1301 on cell proliferation, migration, and angiogenesis.
  • Analysis of miRNA expression in liver cancer cell lines and clinical specimens.
  • Investigation of epigenetic modifications (methylation, histone acetylation) of the miR-1301 gene.

Main Results:

  • Two miRNAs, miR-210 and miR-1301, were identified as specifically targeting KLF6-FL, but not KLF6-SV1.
  • Stable expression of KLF6-FL inhibited cell proliferation, migration, and angiogenesis.
  • Overexpression of miR-1301 promoted cell migration and angiogenesis, indicating an oncogenic role.
  • miR-1301 was found to be highly expressed in liver cancer cell lines and clinical samples.
  • Potential methylation and histone acetylation mechanisms regulating miR-1301 expression were identified.

Conclusions:

  • Specific miRNAs, such as miR-1301, promote liver tumorigenesis by selectively targeting the tumor-suppressive KLF6-FL isoform.
  • This miRNA-mediated targeting of KLF6-FL represents a novel molecular mechanism contributing to cancer progression.
  • The findings highlight the distinct roles of KLF6 isoforms and the potential of targeting miRNA-KLF6 interactions in liver cancer therapy.
Keywords:
Alternative splicingKLF6miRNA

More Related Videos

Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library08:40

Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library

Published on: April 6, 2012

18.5K
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells09:45

An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells

Published on: May 21, 2019

8.1K

Related Concept Videos

MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
3.0K
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After...
21.0K
MicroRNAs01:22

MicroRNAs

9.8K
lncRNA - Long Non-coding RNAs02:39

lncRNA - Long Non-coding RNAs

In humans, more than 80% of the genome gets transcribed. However, only around 2% of the genome codes for proteins. The remaining part produces non-coding RNAs which includes ribosomal RNAs, transfer RNAs, telomerase RNAs, and regulatory RNAs, among other types. A large number of regulatory non-coding RNAs have been classified into two groups depending upon their length – small non-coding RNAs, such as microRNA, which are less than 200 nucleotides in length, and long non-coding RNA...
7.5K
Non-LTR Retrotransposons03:18

Non-LTR Retrotransposons

As the name suggests, non-LTR retrotransposons lack the long terminal repeats characteristic of the LTR retrotransposons. Additionally, both LTR and non-LTR retrotransposons use distinct mechanisms of mobilization. Non-LTR retrotransposons are further divided into two classes - Long interspersed nuclear elements (LINEs) and short interspersed nuclear elements (SINEs), both of which occur abundantly in most mammals, including humans. Some of the active non-LTR retrotransposons in humans are L1...
12.4K