Related Experiment Video
Updated: Apr 14, 2026
Silencing of BRCA2 to Identify Novel BRCA2-regulated Biological Functions in Cultured Human Cells
Published on: August 12, 2015
ShRNA-mediated knock-down of CXCR7 increases TRAIL-sensitivity in MCF-7 breast cancer cells
Weiran Gao1, Xifan Mei2, Jikun Wang1
1Department of Oncology, The First Affiliated Hospital of Liaoning Medical University, No.2, Wuduan, Renmin Street, 121000, Jinzhou, China.
Abstract:
This study aims to investigate the effects of CXCR7-shRNA on TRAIL-mediated apoptosis and suppression of invasive migration and the underlying mechanisms. (1) We constructed CXCR-7-shRNA lentiviral vectors and confirmed their silencing efficiency in MCF-7 cells by RT-PCR analysis. (2) The effects of CXCR7 and/or TRAIL on cell proliferation were examined by MTT assay. (3) Trans well invasion assay was used to examine the effects of CXCR7 silencing and/or TRAIL on MCF-7 cell invasive migration. (4) Expression of Caspase-3, and Caspase-8, and MMP-2 and MMP-9 proteins was examined by Western blot analysis. (1) Viral titers were 2.95 × 10(8) TU/ml, 3.01 × 10(8) TU/ml, 3.26 × 10(8) TU/ml, and 3.08 × 10(8) TU/ml, respectively. (2) CHXR7 shRNAs markedly decreased CXCR7 mRNA expression in MCF-7 cells, among which CXCR7-shRNA-1 showed significantly higher rate of inhibition (P < 0.05). (3) Combination of TRAIL and CXCR7-shRNA-1 resulted in marked suppression of cell proliferation in time-dependent manner (P < 0.05). (4) Cell invasion capacity was inhibited in each experimental group as compared to blank control group at 48 h post treatments (P < 0.05). Among them, combination of TRAIL and CXCR7-shRNA had the highest inhibitory effect (P < 0.05). (5) Western blot analysis indicated that TRAIL alone does not affect CXCR7 expression, but either TRAIL + CXCR7 shRNA or CXCR7 shRNA alone markedly suppressed CXCR7 protein expression. Furthermore, combination of TRAIL and CXCR-7-shRNA significantly increased Caspase-3 and Caspase-8 expression and decreased MMP-2 and MMP-9 expression (P < 0.05). Knock-down of CXCR-7 expression leads to augmented TRAIL-mediated suppression of MCF-7 cell proliferation and invasion.
Insights
This study demonstrates that reducing CXCR7 expression enhances TRAIL-induced apoptosis and inhibits MCF-7 cell proliferation and invasion. Knocking down CXCR7 significantly boosts TRAIL
Area of Science:
- Molecular Biology
- Cancer Research
- Cell Biology
Background:
- CXCR7 (C-X-C chemokine receptor type 7) is implicated in cancer progression.
- TRAIL (TNF-related apoptosis-inducing ligand) is a potent inducer of apoptosis in cancer cells.
- Understanding the interplay between CXCR7 and TRAIL is crucial for developing novel cancer therapies.
Purpose of the Study:
- To investigate the effect of CXCR7-shRNA on TRAIL-mediated apoptosis and invasion in MCF-7 cells.
- To elucidate the underlying molecular mechanisms involving Caspase-3, Caspase-8, MMP-2, and MMP-9.
- To evaluate the combined efficacy of CXCR7 knockdown and TRAIL treatment.
Main Methods:
- Construction and validation of CXCR7-shRNA lentiviral vectors.
- Assessment of cell proliferation using MTT assay.
- Evaluation of cell invasive migration via Transwell invasion assay.
- Analysis of protein expression (Caspase-3, Caspase-8, MMP-2, MMP-9) by Western blot.
Main Results:
- CXCR7-shRNA effectively reduced CXCR7 mRNA and protein expression in MCF-7 cells.
- Combined treatment with TRAIL and CXCR7-shRNA significantly suppressed cell proliferation and invasion.
- The combination therapy markedly increased Caspase-3 and Caspase-8 expression.
- Downregulation of CXCR7 led to decreased expression of MMP-2 and MMP-9.
Conclusions:
- CXCR7 knockdown potentiates TRAIL-induced apoptosis and suppresses invasive migration in MCF-7 cells.
- The combination of TRAIL and CXCR7-shRNA offers a promising therapeutic strategy for targeting breast cancer.
- Modulating CXCR7 expression can enhance the anti-cancer effects of TRAIL.
More Related Videos
10:59Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment
Published on: November 23, 2017
08:59Looking for Driver Pathways of Acquired Resistance to Targeted Therapy: Drug Resistant Subclone Generation and Sensitivity Restoring by Gene Knock-down
Published on: December 11, 2017
