Dysregulated Long Non-Coding RNAs in Colorectal Cancer: Identification and Validation Using RNA-seq and Real-Time Reverse Transcription Polymerase Chain Reaction.

OBJECTIVE

Colorectal cancer (CRC) is the third most common cancer and the second leading cause of cancerrelated deaths worldwide, posing a significant public health challenge. Recent advances in molecular research and technologies have highlighted the potential of long non-coding RNAs (lncRNAs) as key players in CRC development and progression. High-throughput technologies, such as RNA sequencing (RNA-seq), enable comprehensive analysis of cancer omics data, providing valuable insights into CRC biology. This study aimed to identify and validate lncRNAs with potential roles in CRC pathogenesis.

MATERIALS AND METHODS

In this experimental study, we conducted RNA-seq analysis using data from The Cancer Genome Atlas (TCGA) to identify differentially expressed lncRNAs between 481 CRC and 41 healthy control samples. Using the Boruta feature selection algorithm, we identified statistically significant lncRNAs. Based on a comprehensive literature review, four lncRNAs ( and ) were selected for experimental validation. Real-time reverse transcription polymerase chain reaction (RT-PCR) was performed on 10 primary CRC and 9 normal colorectal tissue samples to quantify the expression of these lncRNAs. Additionally, bioinformatic analyses were conducted to explore molecular pathways associated with these lncRNAs in CRC.

RESULTS

RNA-seq analysis identified 388 differentially expressed lncRNAs in CRC tissues compared to healthy controls. Among these, four lncRNAs ( and ) were validated through real-time RT-PCR, confirming their significant upregulation in CRC tissues. Bioinformatic analysis revealed their potential involvement in molecular pathways critical to tumor growth and metastasis.

CONCLUSION

This study identified and validated four significantly upregulated lncRNAs (, and ) in CRC tissues, providing evidence of their potential roles in CRC pathogenesis. These lncRNAs could serve as promising candidates for future biomarker development and therapeutic research in colorectal cancer.