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Published on: March 30, 2019
[Effects of siRNAs targeting CD97 immune epitopes on biological behavior in breast cancer cell line MDA-MB231]
Hua Tian1, Yang Chen1, Jiangang Zhao1
1Department of General Surgery, the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China.
Objective:
To investigate the effects of siRNAs targeting CD97 immune epitopes on proliferation, infiltration, apoptosis and cell cycle of breast cancer cells.
Methods:
siRNA sequences targeting CD97EGF and CD97Stalk immune epitopes were designed according to Gene Bank NM_001025160.2 with smart siCatchTM siRNA design software. CD97siRNAs were transfected into MDA-MB231 cells in which CD97 was highly expressed. Highest sensitive CD97EGF and CD97Stalk siRNA were screened by Western blotting. Inverted microscope was used to observe the growth of CD97siRNAs-transfected MDA-MB231 cells; the proliferation activity of MDA-MB231 cells was detected by MTT method; the wound healing assay and Transwell migration test were performed to examine the migration and infiltration ability of CD97EGF and CD97Stalk siRNA-transfected MDA-MB231 cells; the effects of CD97EGF siRNA and CD97Stalk siRNA on cell apoptosis and cell cycle of MDA-MB231 cells were detected by TUNEL and flow cytometry.
Results:
The growth and proliferation activity of CD97siRNAs-transfected MDA-MB231 cells were significantly lower than those in the control groups, and such differences were more significant in CD97Stalk siRNA-transfected group (all P<0.05); scratch test showed that the wound healing rate was lower in CD97siRNAs-transfected groups, especially in CD97Stalk siRNA-transfected group (all P<0.05); Transwell migration showed that the number of MDA-MB231 cells crossing through chambers were less in CD97siRNAs-transfected groups, especially in CD97Stalk siRNA-transfected group (all P<0.05); no significant difference in cell apoptosis was observed between CD97siRNAs-transfected groups and control groups; cell cycle detection showed that CD97siRNAs-transfected groups had less cells in G0/G1 phase and more cells in S phase compared with the control groups, and such effect on cell cycle was more marked in CD97Stalk siRNA-transfected group (all P<0.05).
Conclusions:
CD97 plays an important role in the cell growth, proliferation, migration and invasion of breast cancer MDA-MB231 cells, and compared with CD97EGF, CD97Stalk may have more effective inhibitory effects on cellular malignant behaviors.
Insights
Small interfering RNAs (siRNAs) targeting CD97 epitopes significantly inhibited breast cancer cell growth and migration. CD97 Stalk siRNA demonstrated more potent effects, impacting cell cycle progression but not apoptosis.
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Silencing
Background:
- CD97 is a protein implicated in cellular processes relevant to cancer.
- Targeting CD97 via RNA interference presents a potential therapeutic strategy for breast cancer.
- Understanding the specific roles of CD97 immune epitopes is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the impact of siRNAs targeting CD97 immune epitopes on breast cancer cell behavior.
- To evaluate the effects on proliferation, migration, invasion, apoptosis, and cell cycle.
- To compare the efficacy of targeting CD97 EGF and CD97 Stalk epitopes.
Main Methods:
- siRNA sequences targeting CD97 EGF and CD97 Stalk epitopes were designed and transfected into MDA-MB231 breast cancer cells.
- Cell proliferation was assessed using MTT assays.
- Migration and invasion were evaluated through wound healing and Transwell assays.
- Apoptosis and cell cycle distribution were analyzed using TUNEL and flow cytometry, respectively.
Main Results:
- CD97 siRNA transfection significantly reduced MDA-MB231 cell growth, proliferation, and migration, with CD97 Stalk siRNA showing a more pronounced inhibitory effect.
- Wound healing and Transwell assays indicated decreased migration and invasion capabilities in siRNA-transfected cells, particularly those treated with CD97 Stalk siRNA.
- While apoptosis rates remained unchanged, cell cycle analysis revealed a shift from G0/G1 to S phase in siRNA-treated cells, more notably with CD97 Stalk siRNA.
Conclusions:
- CD97 plays a significant role in the malignant behaviors of MDA-MB231 breast cancer cells, including growth, proliferation, migration, and invasion.
- CD97 Stalk siRNA exhibits potentially greater inhibitory effects on these cellular behaviors compared to CD97 EGF siRNA.
- Targeting CD97, particularly the Stalk epitope, represents a promising avenue for therapeutic intervention in breast cancer.
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