Abstract
MicroRNAs (miRNAs) are short ~21-nt non-coding RNA molecules that have been shown to regulate a number of biological processes. Previous reports have shown that overexpression of miR-128 in glioma cells inhibited cell proliferation. Literature also suggests that miR-128 negatively regulates prostate cancer cell invasion. Here, we show that overexpression of hsa-miR-128, a brain-enriched microRNA, induces apoptosis in HEK293T cells as elucidated by apoptosis assay, cell cycle changes, loss of mitochondrial membrane potential and multicaspase assay. By in silico analysis, we identified a putative target site within the 3′ untranslated region (UTR) of Bax, a proapoptotic member of the apoptosis pathway. We found that ectopic expression of hsa-miR-128 suppressed a luciferase reporter containing the Bax-3′ UTR and reduced the levels of Bax in HEK293T cells. Taken together, our study demonstrates that overexpression of hsa-miR-128 not only induces apoptosis in HEK293T cells but also is an endogenous regulator of Bax protein.
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Abbreviations
- DHE:
-
Dihydroethidium
- ROS:
-
Reactive oxygen species
- DiOC6:
-
3,3′ Dihexyloxacarbocyanine iodide
- Δψm :
-
Mitochondrial membrane potential
- SR-VAD-FMK:
-
Sulforhodamine-valyl-alanyl-aspartyl-fluoromethyl-ketone
- endoG:
-
Endonuclease G
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Acknowledgments
This work was supported by Grants NWP0036 from the Council of Scientific and industrial research (CSIR). Y.K.A. was supported with fellowship from CSIR. The authors thank Prof. Samir K. Brahmachari for project conceptualization. We also acknowledge Anita Goel, Ravindresh Chhabra, Richa Singh, Richa Dubey and Shruti Chowdhari for their help.
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Adlakha, Y.K., Saini, N. MicroRNA-128 downregulates Bax and induces apoptosis in human embryonic kidney cells. Cell. Mol. Life Sci. 68, 1415–1428 (2011). https://doi.org/10.1007/s00018-010-0528-y
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DOI: https://doi.org/10.1007/s00018-010-0528-y