Related Experiment Video
Updated: Dec 28, 2025
Through the Looking Glass: Time-lapse Microscopy and Longitudinal Tracking of Single Cells to Study Anti-cancer Therapeutics
Published on: May 14, 2016
Downregulation of the Helicase Lymphoid-Specific (HELLS) Gene Impairs Cell Proliferation and Induces Cell Cycle
Xi Liu1, Xuyang Hou1, Yan Zhou1
1Department of General Surgery, Second Xiangya Hospital, Central South University, Changsha, Hunan, People's Republic of China.
Aim:
Colorectal cancer (CRC) is the fourth most frequently diagnosed cancer worldwide. Despite the decrease in mortality of CRC patients, further investigation of the molecular pathogenesis of CRC could unveil new therapeutic targets and offer better prognosis predictions, which might direct attention to epigenetic regulators.
Methods:
Publicly available data from the Gene Expression Omnibus (GEO) database and clinical samples were collected. Bioinformatics methods were used to screen hub genes expressed in CRC. qRT-PCR and Western blotting were used to experimentally determine the expression of one gene of interest, the helicase lymphoid-specific (HELLS) gene, at the RNA and protein levels. Immunohistochemical (IHC) assays were used to correlate the stained HELLS proteins to survival data. Cell proliferation levels were assayed by a CCK-8 kit, a colony formation assay was performed, and flow cytometry was used to quantify the cells at each stage of the cell cycle.
Results:
A total of 225 overlapping genes were screened, including 14 hub genes. Analysis through a protein-protein interaction (PPI) network and the Gene Ontology database was performed by using the Cytoscape and DAVID online tools, respectively. HELLS RNA and protein expression levels in tumor tissues were 2.09-fold higher and 1.46-fold higher, respectively, than in the peritumoral tissues (p < 0.001, p<0.001). HELLS expression was significantly associated with the T stage (p=0.0027), M stage (p=0.0119), and TNM clinical stage (p = 0.0312) and a higher pathological grade (p=0.049). Highly expressed HELLS was reversibly associated with overall survival (log-rank p = 0.027). HELLS siRNA impaired cell proliferation and colony generation in vitro. HELLS siRNA induced significant G2+M arrest in HT29 and HCT116 cells compared with the respective negative controls (82.29% vs 25.85% and 35.41% vs 15.26%, respectively).
Conclusion:
Our data revealed that HELLS was significantly upregulated in CRC and correlated with clinicopathological parameters. High expression of HELLS indicated poor prognosis for CRC patients. HELLS knockdown led to impaired cell proliferation, colony generation, and G2+M cell cycle arrest.
Insights
Helicase lymphoid-specific (HELLS) is upregulated in colorectal cancer (CRC) and linked to poor prognosis. Inhibiting HELLS in CRC cells reduced proliferation and induced cell cycle arrest, suggesting HELLS as a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Epigenetics
Background:
- Colorectal cancer (CRC) remains a leading cause of cancer-related deaths globally.
- Investigating molecular pathogenesis is crucial for identifying novel therapeutic targets and improving prognostic predictions in CRC.
- Epigenetic regulators are increasingly recognized for their role in cancer development.
Purpose of the Study:
- To identify key molecular players in colorectal cancer pathogenesis.
- To investigate the role of the helicase lymphoid-specific (HELLS) gene in CRC.
- To evaluate HELLS as a potential prognostic biomarker and therapeutic target in CRC.
Main Methods:
- Bioinformatic analysis of Gene Expression Omnibus (GEO) data and clinical samples to screen hub genes in CRC.
- Quantitative real-time PCR (qRT-PCR) and Western blotting to assess HELLS RNA and protein expression.
- Immunohistochemical (IHC) assays to correlate HELLS expression with patient survival and clinicopathological parameters.
- In vitro assays (CCK-8, colony formation, flow cytometry) to determine the functional impact of HELLS knockdown on CRC cell proliferation and cell cycle progression.
Main Results:
- HELLS was significantly upregulated in CRC tumor tissues at both RNA (2.09-fold) and protein (1.46-fold) levels compared to peritumoral tissues.
- High HELLS expression correlated significantly with advanced T and M stages, overall TNM clinical stage, and higher pathological grade.
- Elevated HELLS expression was associated with poorer overall survival in CRC patients.
- Knockdown of HELLS using siRNA impaired CRC cell proliferation and colony formation in vitro.
- HELLS inhibition led to significant G2+M cell cycle arrest in HT29 and HCT116 cells.
Conclusions:
- HELLS is significantly overexpressed in colorectal cancer and serves as a potential biomarker for poor prognosis.
- HELLS expression is closely associated with adverse clinicopathological features in CRC patients.
- Targeting HELLS through knockdown inhibits CRC cell proliferation and induces cell cycle arrest, highlighting its potential as a therapeutic target.
Related Concept Videos
02:23Abnormal Proliferation
01:12Role Of Notch Signalling In Intestinal Stem Cell Renewal
Direct cell-to-cell contact is needed for the activation of Notch signaling. The signal is initiated when a notch ligand binds to a receptor on an adjacent cell, also...
01:23Negative Regulator Molecules
02:36DNA Damage can Stall the Cell Cycle
02:36DNA Damage Can Stall the Cell Cycle
02:34Inhibition of Cdk Activity