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Updated: Dec 31, 2025
An In Vitro Protocol for Evaluating MicroRNA Levels, Functions, and Associated Target Genes in Tumor Cells
Published on: May 21, 2019
MiR-765 functions as a tumour suppressor and eliminates lipids in clear cell renal cell carcinoma by downregulating
Wen Xiao1, Cheng Wang1, Ke Chen1
1Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Background:
Lipid accumulation has been highlighted in cancer development and progression, but the exact mechanism remains unclear in renal cell carcinoma (RCC). MicroRNAs (miRNAs) have been confirmed to participate in the pathological processes of cancers, including tumour occurrence and inhibition. However, the role and mechanism of miR-765 have not been elucidated in clear cell renal cell carcinoma (ccRCC).
Methods:
Using The Cancer Genome Atlas (TCGA) database and qRT-PCR, we investigated differences in miR-765 and proteolipid protein 2 (PLP2) expression, as well as their clinical relevance. To investigate the function of miR-765 and PLP2 in ccRCC, we performed in vitro and in vivo experiments to explore their biological functions in ccRCC.
Findings:
In this study, we showed that miR-765 was upregulated in the plasma of ccRCC patients after tumour resection. Consistently, ccRCC tissues had low expression of miR-765 when compared with corresponding non-cancerous tissues. Overexpression of miR-765 suppressed cell proliferation and metastasis in vitro and in vivo. Mechanistic studies demonstrated that PLP2 was a direct target gene of miR-765. PLP2 was highly expressed in ccRCC tissues, and high PLP2 levels were positively correlated with higher tumour stage and grade and poor prognosis. PLP2 expression was negatively correlated with the miR-765 level in patient samples. We further showed that PLP2 restrained the cell metastasis and proliferation induced by miR-765 and reduced the lipid-eliminating effects of miR-765 in renal cancer cells.
Interpretation:
Our findings suggest that miR-765 may function as a tumour suppressor and eliminate lipids in clear cell renal cell carcinoma by targeting PLP2.
Funding:
This work was funded the grants from the National Natural Scientific Foundation of China (Grant No. 81672528, 81672524, 81602218, 31741032, 81902588).
Insights
MicroRNA-765 (miR-765) acts as a tumor suppressor in clear cell renal cell carcinoma (ccRCC). It inhibits cancer cell proliferation and metastasis by targeting proteolipid protein 2 (PLP2), also reducing lipid accumulation.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Lipid accumulation is implicated in renal cell carcinoma (RCC) development, but mechanisms are unclear.
- MicroRNAs (miRNAs) regulate cancer progression; miR-765's role in clear cell RCC (ccRCC) is unknown.
- Understanding miR-765's function is crucial for ccRCC pathogenesis.
Purpose of the Study:
- To investigate the expression and clinical relevance of miR-765 and proteolipid protein 2 (PLP2) in ccRCC.
- To elucidate the functional role and mechanism of miR-765 in ccRCC progression.
- To determine if miR-765 targets PLP2 and influences lipid metabolism in ccRCC.
Main Methods:
- Analysis of The Cancer Genome Atlas (TCGA) database and qRT-PCR for miR-765 and PLP2 expression.
- In vitro and in vivo experiments to assess the functional impact of miR-765 and PLP2.
- Mechanistic studies to identify direct targets and signaling pathways.
Main Results:
- miR-765 was upregulated post-surgery but underexpressed in ccRCC tissues compared to normal tissues.
- Overexpression of miR-765 inhibited ccRCC cell proliferation and metastasis in vitro and in vivo.
- PLP2, a direct target of miR-765, was highly expressed in ccRCC, correlating with poor prognosis and increased tumor stage/grade. PLP2 overexpression counteracted miR-765's effects on proliferation, metastasis, and lipid elimination.
Conclusions:
- miR-765 functions as a tumor suppressor in ccRCC.
- miR-765 exerts its effects by targeting PLP2, inhibiting proliferation and metastasis.
- miR-765 plays a role in lipid metabolism regulation within ccRCC cells.
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