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Differential transcriptome analysis in HPV-positive and HPV-negative cervical cancer cells through CRISPR knockout of

Prakriti Sen1, Pooja Ganguly, Kirti K Kulkarni

  • 1Cancer Biology Lab, School of Biotechnology, KIIT University, Bhubaneswar, India.

Journal of Biosciences
|September 25, 2020
PubMed

Insights

This study reveals how microRNA-214 (miR-214) impacts gene expression in cervical cancer cells. It identifies key genes affected by miR-214 in both HPV-positive and HPV-negative cancer cells.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • Cervical cancer is frequently associated with Human Papillomavirus (HPV) infection.
  • MicroRNAs (miRNAs) play crucial roles in cancer development and progression.
  • miR-214 is recognized as a tumor suppressor microRNA.

Purpose of the Study:

  • To investigate the global gene expression changes induced by miR-214 in HPV-positive and HPV-negative cervical cancer cells.
  • To identify specific genes regulated by miR-214 in the context of HPV status.

Main Methods:

  • Utilized next-generation sequencing (NGS) for RNA sequencing (RNA-seq) analysis.
  • Manipulated miR-214 levels by knockout and overexpression in CaSki (HPV-positive) and C33A (HPV-negative) cell lines.
  • Validated key differentially expressed genes using real-time quantitative PCR (RT-qPCR).

Main Results:

  • Identified 904 upregulated and 365 downregulated genes (fold change +/- 2) between HPV-positive and HPV-negative cells.
  • Observed significant differential expression of 11 genes (e.g., TNFAIP3, MET, CD44) with a fold change of +/- 5.
  • This study is the first to report on the global gene expression effects of miR-214 in cervical cancer concerning HPV status.

Conclusions:

  • miR-214 significantly influences global gene expression in cervical cancer cells, irrespective of HPV status.
  • Specific genes regulated by miR-214 may represent novel therapeutic targets or biomarkers for cervical cancer.
  • Further research into miR-214's role can elucidate mechanisms underlying cervical carcinogenesis and inform treatment strategies.

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