Effects of miR-455 on PIK3R1 gene expression regulation and kidney cancer cell functions
1Basic Medical College of Guizhou Medical University, Guiyang, Guizhou Province, China. zhaolinsunnk@163.com.
Objective:
Migration and proliferation of kidney cancer cells are critical factors affecting effective treatment. Abnormal gene expression exists during cancer pathogenesis. The study showed certain effects of PIK3R1 gene on migration and proliferation of kidney cancer cells, although a few studies have been performed regulatory mechanism of microRNA on PIK3R1 gene. Previous sequencing of PIK3R1 gene found the existence of mir-455 binding site. This study confirmed the regulation of PIK3R1 gene expression in kidney cancer cells of mir-455 by in vitro assay, and analyzed its effects on migration or proliferation of kidney cancer cells.
Materials And Methods:
mir-455 agonist and inhibitor sequences were designed and synthesized to transfect kidney cancer cell line CAKI-1, which were further divided into agonist, inhibitor, and control group. qRT-PCR was used to test expression of mir-455 and PIK3R1 at 12 h, 24 h and 48 h after transfection. PIK3R1 protein expression was measured by Western blot. MTT and cell scratch assay were employed to measure cell proliferation and migration potency.
Results:
12 h after transfection with mir-455, no significant difference of the level of PIK3R1was found among the three groups (p>0.05). However, at 24 h and 48 h post-transfection, PIK3R1 expression in agonist group was significantly elevated, along with weakened cell proliferation or migration potency (p<0.05 with significant between-group comparison). By contrast, the level of PIK3R1 was statistically decreased in inhibitor group, in which cell proliferation and migration were enhanced (p<0.05).
Conclusions:
In kidney carcinoma cell CAKI-1, mir-455 expression regulation can positively alter PIK3R1 gene expression. Over-expression of PIK3Ra gene could reduce the proliferation or migration potency of CAKI-1 cells.
Insights
MicroRNA-455 (mir-455) regulates the PIK3R1 gene in kidney cancer cells. Upregulating mir-455 reduces kidney cancer cell proliferation and migration by increasing PIK3R1 expression.
Area of Science:
- Oncology
- Molecular Biology
- Gene Regulation
Background:
- Kidney cancer cell migration and proliferation are key factors in treatment resistance.
- The PIK3R1 gene is implicated in kidney cancer pathogenesis, but its microRNA regulation is underexplored.
- A binding site for microRNA-455 (mir-455) was identified on the PIK3R1 gene.
Purpose of the Study:
- To investigate the regulatory role of mir-455 on PIK3R1 gene expression in kidney cancer cells.
- To analyze the impact of mir-455-mediated PIK3R1 regulation on kidney cancer cell migration and proliferation.
Main Methods:
- Kidney cancer cell line (CAKI-1) was transfected with mir-455 agonists and inhibitors.
- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to measure mir-455 and PIK3R1 expression levels.
- Western blot analysis assessed PIK3R1 protein expression, while MTT and cell scratch assays evaluated cell proliferation and migration.
Main Results:
- mir-455 transfection affected PIK3R1 expression at 24 and 48 hours post-transfection.
- Upregulation of mir-455 (agonist group) led to increased PIK3R1 expression and decreased cell proliferation and migration.
- Downregulation of mir-455 (inhibitor group) resulted in decreased PIK3R1 expression and enhanced cell proliferation and migration.
Conclusions:
- mir-455 positively regulates PIK3R1 gene expression in kidney carcinoma CAKI-1 cells.
- Modulating PIK3R1 expression via mir-455 can influence kidney cancer cell behavior.
- This study highlights a potential therapeutic target for kidney cancer treatment.
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