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Study on the Biological Characteristics of CD133 (+) Cells Interfered by RNA Interference in Gastric Cancer
Ji-Wei Yu1, Shou-Lian Wang1, Ju-Gang Wu1
1Department of General Surgery, Shanghai 3rd People's Hospital, School of Medicine, Shanghai Jiao-Tong University, No. 280, Mohe Road, Shanghai 201900, China.
Abstract:
Background. To detect the changes of biological characteristics in gastric cancer cells interfered by CD133-specific small interfering RNA (siRNA). Methods. First to select the siRNA which has the strongest interference effect among 3 siRNAs (i.e., siRNA1, siRNA2, and siRNA3) in KATO-III cells by RT-PCR and Western blotting assays. Then, CD133(+) cells were sorted out from KATO-III cells using an immunomagnetic bead sorting method and transfected with the selected siRNA. Furthermore, the proliferating characteristics, the antichemotherapeutic assessment, Transwell invasion assay, monoclonal sphere formation assay, and subcutaneous transplanted tumor formation assay in nude mice were investigated. Results. siRNA3 showed the strongest interference effect in KATO-III cells. As compared to the uninterfered control group, the CD133(+) cells treated by siRNA3 showed significant decreases in the abilities of proliferation, invasion, clone sphere formation, and resistance to antitumour drugs as well as the weight and size of the transplanted tumor, which was nearly similar to that of CD133(-) cells. Additionally, the protein expression level of the EMT factor E-cadherin increased while those of EMT-related Snail and N-cadherin decreased in CD133(+) cells interfered by siRNA3. Conclusion. Inhibition of CD133 gene expression reduces the abilities of gastric cancer cells in proliferation, invasion, clonal sphere formation, and chemoresistance as well as tumor formation in nude mice.
Insights
Small interfering RNA (siRNA) targeting CD133 significantly reduced gastric cancer cell proliferation, invasion, and chemoresistance. This inhibition also decreased tumor formation in mice, highlighting CD133
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Gastric cancer is a significant global health concern.
- CD133 is a cell surface marker implicated in cancer stem cell properties and therapeutic resistance.
- Understanding CD133's role is crucial for developing effective gastric cancer treatments.
Purpose of the Study:
- To investigate the biological impact of inhibiting CD133 expression in gastric cancer cells using small interfering RNA (siRNA).
- To evaluate the effects of CD133 knockdown on gastric cancer cell proliferation, chemoresistance, invasion, and tumor formation.
- To analyze changes in epithelial-mesenchymal transition (EMT) markers following CD133 inhibition.
Main Methods:
- Selection of the most effective CD133-specific siRNA (siRNA3) in KATO-III cells via RT-PCR and Western blotting.
- Isolation of CD133-positive cells using immunomagnetic bead sorting.
- Transfection of sorted cells with siRNA3 and assessment of biological characteristics.
- In vivo studies involving subcutaneous tumor formation in nude mice.
- Analysis of EMT markers including E-cadherin, Snail, and N-cadherin.
Main Results:
- siRNA3 demonstrated the highest efficacy in suppressing CD133 expression in KATO-III cells.
- CD133-positive gastric cancer cells treated with siRNA3 exhibited significantly reduced proliferation, invasion, sphere formation, and chemoresistance.
- Tumor growth, weight, and size in nude mice were substantially decreased after CD133 inhibition.
- siRNA3 treatment led to increased E-cadherin expression and decreased Snail and N-cadherin expression, indicating EMT suppression.
Conclusions:
- Inhibiting CD133 gene expression via siRNA effectively suppresses key malignant behaviors of gastric cancer cells.
- Targeting CD133 holds potential for reducing gastric cancer cell proliferation, invasion, chemoresistance, and tumorigenicity.
- CD133 knockdown influences EMT pathways, offering a potential therapeutic strategy for gastric cancer.
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