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Sequencing Small Non-coding RNA from Formalin-fixed Tissues and Serum-derived Exosomes from Castration-resistant Prostate Cancer Patients
Published on: November 19, 2019
Long Non-Coding RNA AC008972.1 as a Novel Therapeutic Target for Prostate Cancer
Qingqi Zeng1, Jia Liu1, Qijin Wu2
1Department of Pharmacy, Jiangsu Health Vocational College, Nanjing, China.
Abstract:
Prostate cancer is a common male malignancy and the leading cause of cancer death in men. Long non-coding RNAs (lncRNAs), microRNA (miRNAs) and mRNAs networks mediate prostate cancer progression. Herein, we investigated the functions of lncRNA AC008972.1 and its regulatory mechanism in prostate cancer. The expression levels of lncRNA AC008972.1, miR-143-3p, and TAOK2 were detected in prostate cancer tissues and cell lines by reverse transcription-quantitative polymerase chain reaction. PC3 and LNCaP cells were used to establish lncRNA AC008972.1-knockdown, miR-143-3p-overexpressing, and thousand-and-one-amino acid 2 kinase (TAOK2)-downregulated cells. Cell viability was examined by MTT assays and cell proliferation was detected by clone formation assay. Cell migration and invasion were detected by wound scratch assay and transwell chamber assay. The apoptosis rate was analyzed by flow cytometry. The protein expression was detected by Western blot assay. The RNA interaction was explored and validated by RNA binding protein immunoprecipitation (RIP) assay and dual luciferase activity assay. A mouse xenograft model was established to investigate the effect of lncRNA AC008972.1 on prostate cancer progression. High expression of lncRNA AC008972.1 was associated with low overall survival in prostate cancer patients. Downregulation of lncRNA AC008972.1 suppressed prostate cancer progression by inhibiting cell viability, proliferation, migration, and invasion, in addition to the EMT process, whereas cell apoptosis was significantly promoted. LncRNA AC008972.1 bound with miR-143-3p and negatively regulated miR-143-3p expression. MiR-143-3p overexpression suppressed prostate cancer malignant behaviors in vitro. TAOK2 expression was decreased by miR-143-3p through the complementary targeting of TAOK2 mRNA. Downregulation of lncRNA AC008972.1 mitigated prostate cancer malignant behaviors in vitro based on miR-143-3p/TAOK2 node. Furthermore, the data of xenograft model experiment showed that inhibition of lncRNA AC008972.1 suppressed tumor growth in vivo. Knockdown of lncRNA AC008972.1 inhibits prostate cancer cell growth via downregulation of TAOK2 induced by miR-143-3p. LncRNA AC008972.1 acts as an oncogene in the progression of prostate cancer and may provide a novel therapeutic target for prostate cancer.
Insights
Downregulating long non-coding RNA AC008972.1 inhibits prostate cancer progression by affecting the miR-143-3p/TAOK2 pathway, offering a potential new therapeutic target for this malignancy.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Prostate cancer is a leading cause of cancer death in men, with complex regulatory networks involving long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs) mediating its progression.
- Understanding the specific roles and regulatory mechanisms of individual lncRNAs is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the function of lncRNA AC008972.1 in prostate cancer.
- To elucidate the regulatory mechanism of lncRNA AC008972.1 in prostate cancer progression, focusing on its interaction with miR-143-3p and its downstream target, thousand-and-one-amino acid 2 kinase (TAOK2).
Main Methods:
- Quantitative reverse transcription-polymerase chain reaction (RT-qPCR) to detect expression levels of lncRNA AC008972.1, miR-143-3p, and TAOK2.
- Cellular assays including MTT, clone formation, wound scratch, transwell, and flow cytometry to assess viability, proliferation, migration, invasion, and apoptosis.
- RNA immunoprecipitation (RIP) and dual luciferase assays to validate RNA interactions.
- Western blot to detect protein expression.
- Mouse xenograft models to evaluate in vivo tumor growth.
Main Results:
- High expression of lncRNA AC008972.1 correlated with reduced overall survival in prostate cancer patients.
- Downregulation of lncRNA AC008972.1 suppressed prostate cancer cell viability, proliferation, migration, and invasion, while promoting apoptosis and inhibiting epithelial-mesenchymal transition (EMT).
- lncRNA AC008972.1 directly binds to and negatively regulates miR-143-3p, which in turn targets and decreases TAOK2 expression, indicating a regulatory axis of lncRNA AC008972.1/miR-143-3p/TAOK2.
- Inhibition of lncRNA AC008972.1 suppressed tumor growth in vivo.
Conclusions:
- lncRNA AC008972.1 functions as an oncogene in prostate cancer progression.
- Downregulation of lncRNA AC008972.1 inhibits prostate cancer cell growth by reducing TAOK2 expression via the miR-143-3p pathway.
- lncRNA AC008972.1 represents a potential novel therapeutic target for prostate cancer.
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