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Circular RNA has_circ_0000034 accelerates retinoblastoma advancement through the miR-361-3p/ADAM19 axis
Yanhua Jiang1, Fan Xiao1, Lin Wang1
1Department of Ophthalmology, The Fourth People's Hospital of Shenyang, No. 20 Huanghe South Street, Huanggu District, Shenyang, 110031, Liaoning, China.
Insights
Circular RNA hsa_circ_0000034 (circ_0000034) promotes retinoblastoma (RB) growth by sponging microRNA-361-3p (miR-361-3p) and upregulating ADAM19. This suggests circ_0000034 is a potential therapeutic target for RB.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Retinoblastoma (RB) is a common intraocular cancer in young children.
- The role of circular RNA hsa_circ_0000034 (circ_0000034) in RB pathogenesis remains largely unknown.
- Previous studies indicate circ_0000034 is upregulated in RB tissues.
Purpose of the Study:
- To investigate the function and mechanism of circ_0000034 in retinoblastoma.
- To explore the regulatory network involving circ_0000034, microRNA-361-3p (miR-361-3p), and ADAM19 in RB cells.
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) for gene expression analysis.
- Cellular assays (CCK-8, transwell, flow cytometry) to assess RB cell viability, migration, invasion, and apoptosis.
- Dual-luciferase reporter, RNA immunoprecipitation (RIP), and RNA pull-down assays to confirm molecular interactions.
- Western blot analysis for protein level examination.
- In vivo animal experiments to validate functional roles.
Main Results:
- Circ_0000034 expression was significantly elevated in RB tissues and cells.
- Silencing circ_0000034 inhibited RB cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), while promoting apoptosis.
- Circ_0000034 acted as a molecular sponge for miR-361-3p, leading to the upregulation of its target gene, ADAM19.
- Inhibition of miR-361-3p or overexpression of ADAM19 reversed the effects of circ_0000034 knockdown on RB cells.
Conclusions:
- Circ_0000034 promotes RB progression by upregulating ADAM19 via sponging miR-361-3p.
- Circ_0000034 represents a potential therapeutic target for retinoblastoma treatment.
Abstract:
Retinoblastoma (RB) is an intraocular malignancy that mainly occurs in infants and young children under 5 years of age. Circular RNA hsa_circ_0000034 (circ_0000034) was reported to be upregulated in RB tissues. Nevertheless, the function and mechanism of circ_0000034 in RB are unclear. Expression of circ_0000034, microRNA-361-3p (miR-361-3p), and a disintegrin and metalloproteinase 19 (ADAM19) was examined via quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability, migration, invasion, and apoptosis were determined though Cell Counting Kit-8 (CCK-8), transwell, or flow cytometry assays. Caspase-3 activity was detected using a caspase-3 activity assay kit. Some protein levels were examined using Western blot analysis. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, or RNA pull-down assay were performed to verify the relationship between circ_0000034 or ADAM19 and miR-361-3p. The function of circ_0000034 in vivo was confirmed via animal experiment. We verified that circ_0000034 expression was elevated in RB tissues and cells. Circ_0000034 silencing reduced RB growth in vivo, repressed viability, migration, invasion, and EMT, and induced apoptosis of RB cells in vitro. Circ_0000034 acted as a sponge for miR-361-3p, which targeted ADAM19 in RB cells. Furthermore, the inhibition of miR-361-3p restored circ_0000034 knockdown-mediated impacts on viability, migration, invasion, apoptosis, and EMT of RB cells. Moreover, ADAM19 overexpression abolished the influence of miR-361-3p mimic on viability, migration, invasion, apoptosis, and EMT of RB cells. Circ_0000034 expedited RB progression through upregulating ADAM19 via sponging miR-361-3p, which indicated that circ_0000034 might a target for RB therapy.
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