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Circ_0075804 Regulates the Expression of LASP1 by Targeting miR-1287-5p and Thus Affects the Biological Process of
Qichao Han1, Lan Ma1, Li Shao1
1Department of Ophtalmology, Zaozhuang Municipal Hospital Shandong Province, Zaozhuang, Shandong, China.
Purpose:
Increasing evidence reveals that circular RNA (circRNA) dysregulation is involved in retinoblastoma (RB) pathogenesis. To further realize the development of RB, we investigated the role and regulatory mechanism of circ_0075804 in RB.
Methods:
Real-time quantitative PCR (RT-qPCR) and western blot were employed for expression analysis. CCK-8 assay, EdU assay, colony formation assay, flow cytometry assay and transwell assay were performed to monitor cell phenotypes. Xenograft models were established to monitor the role of circ_0075804 on tumor growth. Tumor growth was assessed by the expression of Ki67, N-cadherin, MMP2 and MMP9 via IHC assay. The predicted binding sites between miR-1287-5p and circ_0075804 or LIM and SH3 protein 1 (LASP1) were validated by dual-luciferase reporter assay.
Results:
Upregulation of circ_0075804 and LASP1, and downregulation of miR-1287-5p were shown in RB tissues and cells. Circ_0075804 knockdown repressed RB cell growth, invasion and survival, and hindered tumor development in vivo. MiR-1287-5p was targeted by circ_0075804, and its repression largely reversed the functional effects of circ_0075804 knockdown. LASP1 was a functional target of miR-1287-5p. The inhibition of miR-1287-5p upregulation on RB cell proliferation, survival and invasion was reversed by LASP1 overexpression. Moreover, circ_0075804 knockdown weakened LASP1 expression via increasing miR-1287-5p.
Conclusion:
Circ_0075804 promotes LASP1 expression by targeting miR-1287-5p, thus acting as a contributor to RB carcinogenesis.HighlightsCirc_0075804 is overexpressed in RB.Circ_0075804 knockdown inhibits RB cell malignant phenotypes and tumor growth in vivo.Circ_0075804 regulates RB cell behaviors by targeting miR-1287-5p.MiR-1287-5p affects RB cell behaviors by binding to LASP1.Circ_0075804 regulates LASP1 expression via targeting miR-1287-5p.
Insights
Circular RNA (circRNA) circ_0075804 promotes retinoblastoma (RB) by increasing LASP1 expression via targeting miR-1287-5p. Knockdown of circ_0075804 inhibits RB cell growth and tumor development.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Circular RNAs (circRNAs) are increasingly implicated in the pathogenesis of retinoblastoma (RB).
- Understanding the specific roles of circRNAs in RB is crucial for developing new therapeutic strategies.
Purpose of the Study:
- To investigate the role and regulatory mechanism of circ_0075804 in retinoblastoma (RB).
- To elucidate the molecular pathway involving circ_0075804, miR-1287-5p, and LASP1 in RB progression.
Main Methods:
- Quantitative PCR and Western blot for expression analysis.
- Cellular assays (CCK-8, EdU, colony formation, flow cytometry, Transwell) to assess cell phenotypes.
- Xenograft models and immunohistochemistry (IHC) to evaluate tumor growth and markers.
- Dual-luciferase reporter assays to validate binding sites between circRNAs, miRNAs, and proteins.
Main Results:
- Circ_0075804 and LASP1 were upregulated, while miR-1287-5p was downregulated in RB tissues and cells.
- Knockdown of circ_0075804 suppressed RB cell proliferation, invasion, survival, and tumor growth in vivo.
- Circ_0075804 directly targeted miR-1287-5p, and miR-1287-5p targeted LASP1, mediating the effects of circ_0075804.
Conclusions:
- Circ_0075804 promotes RB carcinogenesis by upregulating LASP1 expression through sponging miR-1287-5p.
- Circ_0075804 serves as a potential therapeutic target for retinoblastoma.
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