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Next, we up- or down-regulated the miR-143 level in human breast cancer cell line MCF-7, aiming to explore the effects of miR-143 on cell growth and the expressions of ERK5 and MAP3K7 in BC.
To investigate the relationship of miR-143 and ERK5, MAP3K7 or cyclin D1, cells were co-transfected with miR-143 mimic/inhibitor and small interfering RNA targeting (si)-ERK5, si-MAP3K7 or si-cyclin D1 (100 nm, Qiagen).
Then, the levels of miR-143, ERK5 and MAP3K7 were detected using SYBR[R] Premix Ex Taq[TM] (TaKaRa, Japan).
Next, sections were blocked with 1% serum, and then incubated with rabbit anti-human ERK5, phospho-ERK5 (p-ERK5), MAP3K7 or phospho-MAP3K7 (p-MAP3K7) polyclonal antibody (1:50, Santa Cruz, USA) at 4[degrees]C overnight.
The membranes were probed with mouse anti-phospho-ERK5 (p-ERK5), ERK5, p-MAP3K7, MAP3K7 and cyclin D1 polyclonal antibodies (1:500, Santa Cruz) and mouse anti-GAPDH monoclonal antibody (1:2000, Sigma) overnight at 4[degrees]C, respectively.
Pearson correlation analysis was used to evaluate the correlation of miR-143 with ERK5 or MAP3K7.
In multiple myeloma, the addition of ERK5 inhibition and the established roles of the JAK-STAT and CDK9 pathways in myeloma cell survival and drug resistance suggest that TG02 could be an exciting addition to the armamentarium for this disease, added Dr Zaknoen.
Based upon the potential utility of inhibiting the ERK5, JAK2 and CDKs pathways, Tragara also intends to initiate a Phase I study of TG02 in patients with solid tumours in the near future.
The company is enthusiastic about TG02's modulation of the ERK5 pathway in solid tumours, said Francis Burrows, head of oncology biology, Tragara.
A second therapeutic program, TG02, is an oral multi-kinase inhibitor that targets the major signaling pathways involving Flt3, JAK2, ERK5 and several cyclin-dependent kinases (CDKs); TG02 is being prepared for IND filing in Q2 2010.
They include extracellular signal-regulated kinase (ERK)-I and -2, p38 kinases, ERK5 (big MAPK1) and c-Jun N-terminal kinases (JNKs) (Liu et al.
Besides ERKI/2, also ERK5 was activated by vitamin C during proliferation (Fig.
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- Erixon, Sigurd
- ERK activator kinase 1
- ERK activator kinase 2
- Erkat, Migran
- Erkat, Migran Armenakovich
- Erkel, Ferenc
- Erklärung von Bern