More importantly, knockdown of B-cell lymphoma 2 abolished the protection of thymosin beta 4 and microRNA-34a inhibitor against advanced glycation endproducts.
The messenger RNA (mRNA) expression levels of miR-34a and B-cell lymphoma-2 (Bcl-2) were detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR).
After 24 h, we evaluated various pro-inflammatory cytokines, metalloproteinases (<i>MMPs</i>), type II collagen (<i>Col2a1</i>), <i>miR-34a, miR-146a, miR-181a</i>, antioxidant enzymes, and B-cell lymphoma (<i>BCL</i><i>)</i><i>2</i> by qRT-PCR, apoptosis and mitochondrial superoxide production by cytometry, p50 nuclear factor (NF)-κB by immunofluorescence.
However, the mRNA expression level of antiapoptotic B-cell lymphoma-2 was markedly decreased by <i>P. undulata</i> treatment.Moreover, <i>P. undulata</i> increased the protein expression of proapoptotic p53 and caspase 3/9 with reducing B-cell lymphoma-2 protein expression level.Thus, <i>P. undulata</i> induced apoptosis in the HepG2 cells by overexpression of miR-34a which regulates p53/B-cell lymphoma-2/caspases signaling pathway.
The results showed that miR‑34a inhibitor significantly upregulated B‑cell lymphoma 2, procaspase‑3, procaspase‑9 and proto‑oncogene c‑Myc protein expression, and downregulated the expression of p21.
The effects of miR-34a on protein levels of B-cell lymphoma 2 (Bcl-2), bcl-2-like protein 4 (Bax), and Wnt/β-catenin pathway-related proteins were evaluated using Western blot analysis.
The protein expression levels of B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein, caspase‑3 and caspase‑9 were determined by western blot analysis, and miR‑34a expression levels were detected using quantitative reverse transcription‑PCR.
The overall shorter survival was strongly related to the abnormal expression of microRNA-34a (miR-34a) and microRNA-206 (miR-206), which target B cell lymphoma-2(Bcl2) and c-Met.
Levels of miR-34a targets, silent information regulator 1 (SIRT1), B-cell lymphoma-2 (Bcl-2), and E2F transcription factor 3 (E2F3), in the cochlea, auditory cortex, and plasma decreased with aging inversely to miR-34a.