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Updated: Sep 2, 2025

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR qPCR
Published on: May 16, 2012
miR-29b-3p inhibits 22Rv1 prostate cancer cell proliferation through the YWHAE/BCL-2 regulatory axis
Jiafu Zhao1,2, Xiaoyan Ma1,3, Houqiang Xu1,2
1Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guizhou University, Guiyang, Guizhou 550025, P.R. China.
Abstract:
Prostate cancer (PCa) is one of the most common malignant tumours in the world and seriously affects health of men. Studies have shown that microRNA (miR)-29b-3p and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein epsilon (YWHAE) play important roles in influencing the proliferation and apoptosis of PCa cells. However, the molecular mechanism of miR-29b-3p and YWHAE in the proliferation and apoptosis of PCa cells remains unclear. In the present study, bioinformatics as well as in vivo and in vitro experiments were used to predict and verify the targeting relationship between YWHAE and mir-29B-3p and investigate the potential roles of YWHAE and mir-29b-3p in the proliferation and apoptosis of 22RV1 cells. Using bioinformatics and a double luciferase system assay, it was confirmed that miR-29b-3p can target YWHAE 3'untranslated region and affect the expression of YWHAE, suggesting that miR-29b-3p may be a potential miRNA of YWHAE. Reverse transcription-quantitative PCR, Cell Counting Kit-8, Transwell and cell scratch assays showed that miR-29b-3p significantly inhibited the proliferation, invasion and migration of 22Rv1 cells (P<0.01). Rescue experiments demonstrated that YWHAE gene introduction reversed the inhibitory effect of miR-29b-3p on 22Rv1 cells. Western blotting revealed that the upregulation of miR-29b-3p inhibited YWHAE expression, resulting in a very significant decrease in the ratio of p-BAD/BAD and full-length caspase 3/cleaved caspase 3 (P<0.01) and an extremely significant increase in the ratio of BAX/BCL-2 (P<0.01). A tumourigenesis test in nude mice in vivo confirmed that the upregulation of miR-29b-3p inhibited tumour growth by targeting YWHAE. The present experiments confirmed that miR-29b-3p plays a tumour suppressor role in 22Rv1 PCa cells, and the YWHAE/BCL-2 regulatory axis plays a vital role in miR-29b-3p regulating the proliferation and apoptosis of 22Rv1 cells. These results may provide a theoretical basis for the diagnosis and targeted treatment of PCa.
Insights
MicroRNA (miR)-29b-3p acts as a tumor suppressor in prostate cancer (PCa) by inhibiting proliferation and inducing apoptosis. It targets YWHAE, impacting the YWHAE/BCL-2 axis for PCa cell regulation.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Prostate cancer (PCa) is a leading cause of cancer-related deaths in men worldwide.
- MicroRNA (miR)-29b-3p and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein epsilon (YWHAE) are implicated in PCa cell proliferation and apoptosis.
- The precise molecular mechanisms linking miR-29b-3p and YWHAE in PCa remain incompletely understood.
Purpose of the Study:
- To investigate the targeting relationship between YWHAE and miR-29b-3p.
- To elucidate the roles of YWHAE and miR-29b-3p in the proliferation and apoptosis of 22RV1 PCa cells.
- To explore the potential of miR-29b-3p as a therapeutic target for PCa.
Main Methods:
- Bioinformatics analysis and dual-luciferase reporter assays to confirm the targeting interaction.
- In vitro assays including RT-qPCR, CCK-8, Transwell, and cell scratch assays to assess cell proliferation, invasion, and migration.
- In vivo tumor xenograft models in nude mice to evaluate tumor growth inhibition.
Main Results:
- miR-29b-3p directly targets the 3' untranslated region of YWHAE, downregulating its expression.
- Upregulation of miR-29b-3p significantly inhibited proliferation, invasion, and migration of 22RV1 cells.
- miR-29b-3p suppressed tumor growth in vivo by targeting YWHAE, decreasing p-BAD/BAD and caspase 3 ratios, and increasing BAX/BCL-2 ratio.
Conclusions:
- miR-29b-3p functions as a tumor suppressor in 22RV1 PCa cells.
- The YWHAE/BCL-2 regulatory axis is crucial for miR-29b-3p-mediated regulation of proliferation and apoptosis in PCa.
- These findings offer a potential theoretical basis for PCa diagnosis and targeted therapy.
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