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In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice
Published on: August 23, 2019
Vector-based miR-15a/16-1 plasmid inhibits colon cancer growth in vivo
Lixia Dai1, Wei Wang, Shuang Zhang
1State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan, People's Republic of China.
Abstract:
miR-15 (microRNA 15) and miR-16 are frequently deleted or down-regulated in many cancer cell lines and various tumour tissues, suggesting that miR-15a/16-1 plays important roles in tumour progression and might be a method for cancer treatment. We have developed a vector-based plasmid to explore the anti-tumour efficacy of miR-15a/16-1 in colon cancer in vivo. It is proposed that miR-15a and miR-16-1 target cyclin B1 (CCNB1), which associates with several tumorigenic features such as survival and proliferation. The levels of miR-15a and miR-16-1 in colon cancer cells were inversely correlated with CCNB1 expression, and there was consensus between miR-15a/16-1 and CCNB1 mRNA sequences by analysing homology. Vector-based miR-15a/16-1 expression plasmid was constructed and transfected into HCT 116 and SW620 colon cancer cells in vitro. The effects produced on cell viability and angiogenesis were analysed using flow cytometric analysis, colony formation analysis and tube formation analysis. CCNB1 expression down-regulation was checked by Western blotting. Systemic delivery of miR-15a/16-1 plasmids encapsulated in cationic liposome led to a significant inhibition of subcutaneous tumour growth and angiogenesis in tumour tissues, whereas no effects were observed with liposome carrying the non-specific plasmid. In summary, miR-15a/16-1 has been applied in colon cancer treatment in vivo, and resulted in effective colon tumour xenografts growth arrest and angiogenesis decrease. These findings suggest that systemic delivery of vector-based miR-15a/16-1 expression plasmid can be an approach to colon cancer therapy.
Insights
MicroRNA-15a/16-1 (miR-15a/16-1) effectively inhibits colon cancer growth and angiogenesis by targeting cyclin B1 (CCNB1). Systemic delivery of miR-15a/16-1 plasmids offers a promising therapeutic strategy for colon cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- MicroRNAs (miRNAs), specifically miR-15a/16-1, are frequently downregulated in various cancers, indicating their tumor-suppressive roles.
- Cyclin B1 (CCNB1) is a key regulator of cell cycle progression and is often overexpressed in tumors, correlating with poor prognosis.
- The inverse correlation between miR-15a/16-1 and CCNB1 expression in colon cancer suggests a direct regulatory relationship.
Purpose of the Study:
- To investigate the anti-tumor efficacy of miR-15a/16-1 in colon cancer using a vector-based plasmid system.
- To evaluate the impact of miR-15a/16-1 on colon cancer cell viability, proliferation, and angiogenesis in vitro and in vivo.
- To confirm the targeting of CCNB1 by miR-15a/16-1 and its role in mediating anti-tumor effects.
Main Methods:
- Construction and transfection of a vector-based miR-15a/16-1 expression plasmid into colon cancer cell lines (HCT 116, SW620).
- In vitro assays including flow cytometry, colony formation, and tube formation to assess cell viability and angiogenesis.
- Western blotting to confirm CCNB1 expression down-regulation.
- In vivo studies involving systemic delivery of miR-15a/16-1 plasmids encapsulated in cationic liposomes to evaluate tumor growth inhibition and angiogenesis in subcutaneous xenografts.
Main Results:
- Vector-based miR-15a/16-1 expression significantly reduced colon cancer cell viability and angiogenesis in vitro.
- Down-regulation of CCNB1 expression was confirmed by Western blotting, consistent with miR-15a/16-1 targeting.
- Systemic administration of miR-15a/16-1 plasmids effectively inhibited subcutaneous colon tumor growth and angiogenesis in vivo.
- Control liposomes carrying non-specific plasmids showed no significant anti-tumor effects.
Conclusions:
- miR-15a/16-1 exhibits potent anti-tumor activity against colon cancer by suppressing cell proliferation and angiogenesis.
- Targeting CCNB1 by miR-15a/16-1 is a key mechanism underlying its therapeutic effect.
- Systemic delivery of vector-based miR-15a/16-1 expression plasmids represents a viable and effective strategy for colon cancer therapy.
