Related Experiment Video
Updated: Aug 9, 2026
The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers
Published on: January 22, 2013
[Screening of novel genes differentially expressed in human renal cell carcinoma by suppression subtractive
Jun-kui Ai1, Xiao Huang, Yu-juan Wang
1Institute of Urology, First Hospital, Peking University, Beijing 100034, P. R. China.
Background And Objective:
Identifying the differentially expressed genes in renal cell carcinoma (RCC) contributes to the elucidation of its genetic basis. However, the above knowledge has not yet been fully understood. The aims of this experiment were to screen novel genes differentially expressed in RCC tissues by suppression subtractive hybridization (SSH) and clone RCC-specific related genes.
Methods:
To construct SSH library of RCC by using the mRNA from RCC tissues and matched normal kidney tissues as tester and driver, respectively. Partial positive clones in the library were selected randomly and sequenced, then analyze the sequences with the BLAST software. To confirm the location of the fragments of interest in human chromosome through comparing their sequences with the human genome draft. mRNA levels of the novel genes in RCC and matched normal kidney tissues were determined by Northern blot and semi-quantitative RT-PCR analysis.
Results:
The SSH library contained 414 positive clones. Random analysis of 280 clones with enzyme restriction showed that 265 clones contained cDNA fragments distributed mainly between 300-900bp. Among 80 arbitrary clones with were derived from above 265 clones and sequenced, No. 28, 158, 170, and 249 clones are previously unknown genes and located in human chromosome 21q22, 4p15.3, 9q34, and 22q11.2 by electronic mapping, respectively. The consequence of semi-quantitative RT-PCR demonstrated that mRNA levels of the two novel genes were overexpressed in RCC compared to matched normal tissues by more than 2-6 folds. Northern blot analysis confirmed the above results.
Conclusions:
SSH is a reliable strategy for screening novel genes differentially expressed in RCC. The novel gene fragments can be used to clone their full length and further to study their functions.
Insights
This study identified novel genes overexpressed in renal cell carcinoma (RCC) using suppression subtractive hybridization (SSH). These findings advance understanding of RCC's genetic basis and potential therapeutic targets.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Context:
- Renal cell carcinoma (RCC) is a significant health concern, and understanding its genetic underpinnings is crucial for diagnosis and treatment.
- Previous research has not fully elucidated the genetic landscape of differentially expressed genes in RCC.
Purpose:
- To screen for novel genes exhibiting differential expression in RCC tissues using suppression subtractive hybridization (SSH).
- To clone and characterize RCC-specific genes for further functional studies.
Summary:
- Suppression subtractive hybridization (SSH) was employed to construct a library from RCC and normal kidney tissues.
- Analysis identified four previously unknown genes located on human chromosomes 21q22, 4p15.3, 9q34, and 22q11.2.
- Semi-quantitative RT-PCR and Northern blot confirmed overexpression of two novel genes in RCC tissues (2-6 fold increase).
Impact:
- SSH proves to be a reliable method for identifying novel differentially expressed genes in RCC.
- The identified gene fragments serve as a basis for cloning full-length genes and investigating their roles in RCC pathogenesis.
- This research contributes to the elucidation of RCC's genetic basis and may reveal new diagnostic or therapeutic targets.
More Related Videos
09:44High-throughput Screening for Chemical Modulators of Post-transcriptionally Regulated Genes
Published on: March 3, 2015
09:33Genetic Profiling and Genome-Scale Dropout Screening to Identify Therapeutic Targets in Mouse Models of Malignant Peripheral Nerve Sheath Tumor
Published on: August 25, 2023
