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Long noncoding RNA GPRC5D-AS1 in renal cell carcinoma: a molecular mechanism study
Mingfei Jia1, Xi Chen2, Shaosan Kang2
1North China Graduate School of University of Science and Technology, Tangshan, China.
Background:
Clear cell renal cell carcinoma (RCC) is the most common subtype of RCC. Although targeted therapy can provide superior treatment outcomes, it is prone to drug resistance, and individual responses to immunotherapy vary greatly. Therefore, finding new diagnostic and therapeutic targets for RCC is of considerable importance. Long noncoding RNA (lncRNA) GPRC5D-AS1 can serve as a biomarker in clinical applications and the prognosis of lung squamous cell carcinoma. However, the specific mechanism of action of lncRNA GPRC5D-AS1 in RCC has not yet been clarified. Therefore, this paper explores the expression of lncRNA GPRC5D-AS1 in the renal cancer cell line 786-0, and conducts a preliminary study of its molecular mechanism. Selecting nude mice for tumor experiments is because of the high genomic and physiological similarity between mice and humans. Conducting tumor research on mice allows for better control of experimental conditions, aiding researchers in more accurately observing and analysing tumor characteristics and responses.
Methods:
Small interfering RNA (siRNA) and plasmid cloning DNA (pcDNA) 3.1 were used to transfect renal cancer cell line 786-0 to silence and overexpress the lncRNA GPRC5D-AS1 gene. Quantitative real-time fluorescence polymerase chain reaction was used to detect the difference in lncRNA GPRC5D-AS1 expression in blank control group, negative control group, siGPRC5D-AS1 group and oeGPRC5D-AS1 group. The effects of silence and overexpression of lncRNA GPRC5D-AS11 on the proliferation of 786-0 cells were detected in cell colony formation experiments; the changes in the migration and invasion of 786-0 cells were detected via cell scratch assay and transwell assay, respectively; the differences in tumor growth between groups were determined via tumorigenesis experiments in nude mice; and the expression of proliferation-related protein [β-catenin, Ki67 and proliferating cell nuclear antigen (PCNA)] and invasion-related protein (N-cadherin and E-cadherin) were detected via Western blotting.
Results:
Compared with blank control group and negative control group, the siGPRC5D-AS1 group showed a significant decrease in the relative expression of lncRNA GPRC5D-AS1 (P<0.05), a significant increase in the number of proliferating cells and migrating cells (P<0.05), a significant increase in the tumor volume of nude mice (P<0.05), a significant increase in β-catenin, Ki67, PCNA and N-cadherin protein expression (P<0.05), and a significant decrease in E-cadherin protein expression (P<0.05); conversely, these results were opposite for the eGPRC5D-AS1 group.
Conclusions:
Silencing the expression of lncRNA GPRC5D-AS1 can enhance the proliferation, invasion, and migration ability of renal cancer cell line 786-0, which can be weakened by the overexpression of lncRNA GPRC5D-AS1.
Insights
Silencing long noncoding RNA GPRC5D-AS1 enhances renal cell carcinoma cell proliferation and invasion. Conversely, overexpressing GPRC5D-AS1 weakens these aggressive traits, suggesting its potential as a therapeutic target.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Clear cell renal cell carcinoma (RCC) is a prevalent cancer subtype.
- Existing therapies like targeted therapy and immunotherapy face challenges with drug resistance and variable patient responses.
- Identifying novel diagnostic and therapeutic targets for RCC is crucial.
Purpose of the Study:
- To investigate the role and molecular mechanism of long noncoding RNA (lncRNA) GPRC5D-AS1 in renal cell carcinoma (RCC).
- To explore the potential of lncRNA GPRC5D-AS1 as a biomarker or therapeutic target in RCC.
Main Methods:
- Transfection of the 786-0 renal cancer cell line with siRNA and pcDNA to silence and overexpress lncRNA GPRC5D-AS1.
- Quantitative real-time fluorescence polymerase chain reaction (qRT-PCR) to assess gene expression.
- Cell proliferation assays (colony formation), migration assays (scratch assay), and invasion assays (Transwell assay).
- Tumorigenesis experiments in nude mice and Western blotting for proliferation and invasion-related proteins.
Main Results:
- Silencing GPRC5D-AS1 significantly increased cell proliferation, migration, and invasion in 786-0 cells.
- Tumor growth in nude mice was significantly increased upon GPRC5D-AS1 silencing.
- Protein analysis revealed increased expression of proliferation markers (β-catenin, Ki67, PCNA) and an invasion marker (N-cadherin), with decreased E-cadherin expression after GPRC5D-AS1 silencing.
- Conversely, overexpression of GPRC5D-AS1 showed opposite effects.
Conclusions:
- Silencing lncRNA GPRC5D-AS1 promotes proliferation, invasion, and migration in renal cancer cells.
- Overexpression of lncRNA GPRC5D-AS1 inhibits these aggressive behaviors.
- lncRNA GPRC5D-AS1 plays a significant role in RCC progression and may serve as a potential therapeutic target.
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