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Published on: March 30, 2019
The Expression of miR-34c-5p Induces G0/G1 Cell Cycle Arrest and Apoptosis in SW480 Colon Cancer Cell
Shirin Farzaneh1, Shabnam Bandad1, Faezeh Shaban1
1Pharmaceutical Sciences Center, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran.
Background:
Expression of the miR-34 family, including miR-34a/b/c, has been reported to inhibit the progression of several cancer types by inhibiting cell proliferation and inducing apoptosis.
Objectives:
We attempted to investigate the effect of SW480 cell transfection with miR-34c-5p mimics on cell proliferation.
Methods:
To do this, SW480 colon cancer cell line was transfected with miR-34c-5p mimics, scramble sequence, and the vehicle in PBS mock, and then cell proliferation was assessed by MTT assay. The population of cells in cell cycle phases, ROS generation, and apoptosis rate were evaluated by flow cytometry. Additionally, we determined the relative expression of apoptotic genes through real-time PCR technique.
Results:
We observed a reduced proliferation rate in cells transfected with miR-34c-5p compared to the control group (P <0.05). We also found that miR-34c-5p caused a significant increase in apoptosis rate (P < 0.001) and cell cycle arrest in the G0 and G1 phases (P < 0.05). Moreover, a significant increase was reported in the expression of pro-apoptotic genes, including BAK (P < 0.001), BAX and BAD (P < 0.0001), and Caspase 7/9 (P < 0.0001).
Conclusions:
However, no remarkable difference was seen in the expression of MCL1, BCL2, and CASPASE 3 genes. Our conclusion is that overexpression of miR-34c-5p could be considered a promising approach for colorectal cancer treatment.
Insights
Overexpression of miR-34c-5p mimics significantly reduced colon cancer cell proliferation and increased apoptosis. This microRNA shows promise as a therapeutic agent for colorectal cancer treatment.
Area of Science:
- Molecular Biology
- Oncology
- Gene Regulation
Background:
- The miR-34 family, including miR-34a/b/c, is known to inhibit cancer progression.
- These microRNAs function by suppressing cell proliferation and promoting apoptosis.
Purpose of the Study:
- To investigate the impact of miR-34c-5p mimic transfection on SW480 colon cancer cell proliferation.
- To evaluate the effects of miR-34c-5p on apoptosis and cell cycle progression.
Main Methods:
- SW480 colon cancer cells were transfected with miR-34c-5p mimics, a scramble sequence, or a vehicle control.
- Cell proliferation was assessed using MTT assay.
- Apoptosis rates, cell cycle distribution, and apoptotic gene expression (BAK, BAX, BAD, Caspase 7/9) were analyzed via flow cytometry and real-time PCR.
Main Results:
- Transfection with miR-34c-5p mimics led to a significant reduction in cell proliferation.
- A notable increase in apoptosis rate and G0/G1 cell cycle arrest was observed.
- Expression of pro-apoptotic genes BAK, BAX, BAD, and Caspase 7/9 was significantly upregulated.
Conclusions:
- Overexpression of miR-34c-5p effectively inhibits colon cancer cell proliferation and induces apoptosis.
- miR-34c-5p represents a potential therapeutic strategy for colorectal cancer.
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