生物活性 |
CMLD-2, an inhibitor of HuR-ARE interaction, competitively binds HuR protein disrupting its interaction with adenine-uridine rich elements (ARE)-containing mRNAs (Ki=350 nM). CMLD-2 induces apoptosis exhibits antitumor activity in different cancer cells as colon, pancreatic, thyroid and lung cancer cell lines. Hu antigen R (HuR) is an RNA binding protein, can regulate target mRNAs stability and translation.
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体外研究 |
CMLD-2 (1-75 μM; 24-72 h) inhibits thyroid cancer cell viability.
CMLD-2 (20-30 μM; 24-48 h) activates caspases and induces apoptotic cell death in H1299 and A549 cells.
CMLD-2 (30 μM; 24-48 h) induces G1 cell cycle arrest and mitochondrial perturbation in H1299 and A549 cell.
CMLD-2 (30 μM; 24-48 h) reduces expression of HuR and HuR-regulated mRNAs and proteins in H1299 cells.
CMLD-2 (35 μM; 72 h) decreases directional migration capability in SW1736, 8505C, BCPAP and K1 cells. CMLD-2 induces a strong decrease of MAD2 mRNA levels in SW1736, 8505C, BCPAP and K1 cells.
Cell Viability Assay
Cell Line: |
SW1736, 8505C, BCPAP and K1 cells |
Concentration: |
1, 5, 10, 25, 35, 50, 75 μM |
Incubation Time: |
24, 48, 72 hours |
Result: |
Reduced the viability of all the four cell lines when used at 35, 50 and 75 μM concentration and at different time points. |
Apoptosis Analysis
Cell Line: |
H1299, A549, H1975, HCC827, MRC-9 and CCD16 cells |
Concentration: |
20, 30 μM |
Incubation Time: |
24, 48 hours |
Result: |
Marked activated the caspase-9 and -3 in lung tumor cells.
Induce the cleavage of PARP in lung tumor cells.
Significantly increased the annexin-V-positive staining in lung tumor cells.
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Cell Cycle Analysis
Cell Line: |
H1299, A549, MRC-9 and CCD16 cells |
Concentration: |
30 μM |
Incubation Time: |
24, 48 hours |
Result: |
Induced greater G1 phase cell cycle arrest in H1299 and A549 cells than in MRC-9 and CCD16 cells.
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Western Blot Analysis
Cell Line: |
H1299, A549, H1975, HCC827, CCD16 and MRC-9 cells |
Concentration: |
20, 30 μM |
Incubation Time: |
24, 48 hours |
Result: |
Diminished protein expression of HuR, Bcl-2, Cyclin E and Bcl-XL and increased expression of p27 and BAX in lung tumor cells.
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