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Targeting CRMP-4 by lentivirus-mediated RNA interference inhibits SW480 cell proliferation and colorectal cancer
Si-Le Chen1, Shi-Rong Cai1, Xin-Hua Zhang1
1Department of Gastrointestinal and Pancreatic Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China.
Abstract:
The aim of the present study was to investigate the expression level of collapsin response mediator protein 4 (CRMP-4) in human colorectal cancer (CRC) tissue and to evauluate its impact on SW480 cell proliferation, in addition to tumor growth in a mouse xenograft model. Clinical CRC tissue samples were collected to detect the CRMP-4 protein expression levels using western blot and immunohistochemistry analyses. A specific small interfering RNA sequence targeting the CRMP-4 gene (DPYSL3) was constructed and transfected into an SW480 cell line using a lentivirus vector to obtain a stable cell line with low expression of CRMP-4. The effectiveness of the interference was evaluated using western blot and reverse transcription-quantitative polymerase chain reaction, and the cell proliferation was determined using MTT and BrdU colorimetric methods. Tumor growth was assessed by subcutaneously inoculating the constructed cells into BALB/c nude mice. The protein expression levels of CRMP-4 were markedly increased in colon tumor tissue of the human samples. The proliferation of SW480 cells and the tumor growth rate in nude mice of the si-CPMR-4 group were evidently depressed compared with the si-scramble group. Thus, the present results suggest that CRMP-4 may be involved in the pathogenesis of CRC.
Insights
Collapsin response mediator protein 4 (CRMP-4) is elevated in colorectal cancer (CRC) tissues. Reducing CRMP-4 expression suppressed CRC cell proliferation and tumor growth, suggesting its role in CRC development.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Colorectal cancer (CRC) is a significant global health concern.
- Understanding the molecular mechanisms driving CRC progression is crucial for developing effective therapies.
Purpose of the Study:
- To investigate the expression of collapsin response mediator protein 4 (CRMP-4) in human CRC.
- To evaluate the impact of CRMP-4 on colorectal cancer cell proliferation and tumor growth.
Main Methods:
- Western blot and immunohistochemistry were used to detect CRMP-4 expression in clinical CRC samples.
- CRMP-4 gene (DPYSL3) was silenced in SW480 cells using small interfering RNA (siRNA) delivered via lentivirus.
- Cell proliferation was assessed using MTT and BrdU assays.
- Tumorigenicity was evaluated in a mouse xenograft model.
Main Results:
- CRMP-4 protein levels were significantly increased in human colorectal tumor tissues compared to normal tissues.
- Silencing CRMP-4 in SW480 cells led to reduced cell proliferation in vitro.
- Tumor growth in the mouse xenograft model was significantly inhibited in the CRMP-4-silenced group.
Conclusions:
- CRMP-4 is overexpressed in colorectal cancer.
- CRMP-4 plays a role in promoting colorectal cancer cell proliferation and tumor growth.
- CRMP-4 may serve as a potential therapeutic target for colorectal cancer.
