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Published on: October 30, 2013
Mir-135a enhances cellular proliferation through post-transcriptionally regulating PHLPP2 and FOXO1 in human bladder
Xiao Peng Mao1, Luo Sheng Zhang2, Bin Huang3
1Department of Urology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, PR China. mxpzc1979@163.com.
Background:
Bladder cancer is the most common malignancy in urinary system and the ninth most common malignancy in the world. MicroRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression by targeted repression of transcription and translation and play essential roles during cancer development. We investigated the expression of miR-135a in bladder cancer and explored its bio-function during bladder cancer progression.
Methods:
The expression of miR-135a in bladder cancer cells and tissues are performed by using Real-time PCR assay. Cell viability assay (MTT assay), colony formation assay, anchorage-independent growth ability assay and Bromodeoxyuridine labeling and immunofluorescence (BrdUrd) assay are used to examine cell proliferative capacity and tumorigenicity. Flow cytometry analysis is used to determine cell cycle progression. The expressions of p21, p27, CyclinD1, Ki67, PHLPP2 and FOXO1 are measured by Western blotting assay. Luciferase assay is used to confirm whether FOXO1 is the direct target of miR-135a.
Results:
miR-135a is upregulated in bladder cancer cells and tissues. Enforced expression of miR-135a promotes bladder cancer cells proliferation, whereas inhibition of miR-135a reverses the function. Furthermore, for the first time we demonstrated PHLPP2 and FOXO1 are direct targets of miR-135a and transcriptionally down-regulated by miR-135a. Suppression of PHLPP2 or FOXO1 by miR-135a, consisted with dysregulation of p21, p27, Cyclin D1 and Ki67, play important roles in bladder cancer progression.
Conclusion:
Our study demonstrates that miR-135a promotes cell proliferation in bladder cancer by targeting PHLPP2 and FOXO1, and is performed as an onco-miR.
Insights
MicroRNA-135a (miR-135a) is upregulated in bladder cancer, promoting cell proliferation by targeting PHLPP2 and FOXO1. This study identifies miR-135a as a potential onco-microRNA in bladder cancer progression.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Bladder cancer is a prevalent malignancy globally.
- MicroRNAs (miRNAs) are key regulators in cancer development.
- Investigating specific miRNAs like miR-135a is crucial for understanding bladder cancer progression.
Purpose of the Study:
- To determine the expression levels of miR-135a in bladder cancer.
- To elucidate the functional role of miR-135a in bladder cancer cell proliferation and tumorigenicity.
- To identify the direct molecular targets of miR-135a in bladder cancer.
Main Methods:
- Real-time PCR for miR-135a expression analysis.
- Cell proliferation and tumorigenicity assays (MTT, colony formation, anchorage-independent growth, BrdUrd).
- Western blotting for target gene expression and luciferase assay for target validation.
Main Results:
- miR-135a is significantly upregulated in bladder cancer tissues and cells.
- Overexpression of miR-135a enhances bladder cancer cell proliferation; inhibition reduces it.
- PHLPP2 and FOXO1 were identified as direct targets of miR-135a, mediating its oncogenic effects.
Conclusions:
- miR-135a acts as an onco-microRNA in bladder cancer.
- miR-135a promotes bladder cancer cell proliferation by downregulating PHLPP2 and FOXO1.
- Targeting miR-135a presents a potential therapeutic strategy for bladder cancer.
