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MiR-335 functions as a tumor suppressor and regulates survivin expression in osteosarcoma
1Department of Orthopedics, Xinjiang Uygur Autonomous Region Chinese Medicine Hospital, Urumqi, China. zhaojiang5568@163.com.
Objective:
Previous studies have shown that miR-335 plays an anti-tumor role in several types of cancer. However, whether it is able to regulate the tumorigenesis of osteosarcoma (OS) has not been fully investigated. The present study was designed to study its potential role in regulating apoptosis of OS cells.
Materials And Methods:
The expression of miR-335 in a total of 18 paired OS tumor tissues and adjacent non-cancerous tissues was measured by Real-time PCR, and its different expression in OS cell lines was also measured. The effect of miR-335 on apoptosis was measured by MTT assay, caspase-3 activity assay and TUNEL assay. The effect of survivin inhibition on apoptosis of OS cells was determined by MTT assay and western blot. Luciferase reporter assay and western blot were conducted to confirm the relationship between miR-335 and the 3'UTR of survivin mRNA.
Results:
MiR-335 expression was found to be significantly downregulated in OS tumor tissues and OS cell lines. Overexpression of miR-335 led to decreased cell viability and increased apoptosis. MiR-335 directly targeted the 3'UTR of survivin mRNA and suppressed survivin gene expression, and inhibition of survivin exhibited similar effects to miR-335 overexpression.
Conclusions:
MiR-335 might function as a tumor suppressor in OS, and downregulation of miR-335 in OS cells contributes to the decreased apoptotic potential of OS cells through derepression of survivin.
Insights
MicroRNA-335 (miR-335) acts as a tumor suppressor in osteosarcoma (OS). Its downregulation in OS cells reduces apoptosis by increasing survivin expression, highlighting miR-335 as a potential therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- MicroRNA-335 (miR-335) demonstrates anti-tumor properties in various cancers.
- The specific role of miR-335 in osteosarcoma (OS) tumorigenesis remains underexplored.
- Investigating miR-335's function in OS apoptosis is crucial for understanding cancer progression.
Purpose of the Study:
- To elucidate the role of miR-335 in regulating osteosarcoma cell apoptosis.
- To determine the expression levels of miR-335 in OS tissues and cell lines.
- To identify the molecular mechanisms underlying miR-335's function in OS.
Main Methods:
- Quantitative real-time PCR to measure miR-335 expression in OS tissues and cell lines.
- MTT, caspase-3 activity, and TUNEL assays to assess apoptosis.
- Western blot and luciferase reporter assays to investigate the interaction between miR-335 and survivin mRNA.
Main Results:
- miR-335 expression was significantly reduced in osteosarcoma tissues and cell lines.
- Overexpression of miR-335 inhibited OS cell viability and promoted apoptosis.
- miR-335 directly targets the 3' untranslated region of survivin mRNA, suppressing its expression.
Conclusions:
- miR-335 functions as a tumor suppressor in osteosarcoma.
- Downregulation of miR-335 contributes to reduced apoptosis in OS cells via survivin derepression.
- miR-335 represents a potential therapeutic target for osteosarcoma treatment.
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