Cell lines

MPNST cells

Preparation Method

MPNST cells were starved overnight, incubated with BCI (2 uM) for 60 mins then stimulated with DMEM and 10% FBS for 1 hr. Immunoblot analysis of TP53, p-RB and p-ATM and PARP cleavage and CC3 in ST8814 and S462.TY MPNST cells 24h after treatment with BCI (2 uM).

Reaction Conditions

2 uM; 60 mins

Applications

After 1 hr, p-ERK, p-JNK, p-c-jun and total c-jun were elevated in the BCI-treated MPNST cell lines ST8814 and S462.TY but did not change in iHSC-1λ. Within 24 hours, BCI decreased total PARP and increased cleaved PARP and CC3, indicative of apoptotic cell death in NF1 deficient ST8814 and S462.TY cells.

Animal models

Female C57BL/6 mice (8-weeks old)

Dosage form

15 mg/kg or 30 mg/kg; i.p.

Preparation method

Low- or high-concentration (15 mg/kg or 30 mg/kg) BCI was injected intraperitoneally for 8 weeks, and bone loss was evaluated by micro-CT.

Applications

Bone loss was prevented in both the low- and high-concentration BCI groups. Moreover, quantitative results indicated obvious increases in bone volume/total tissue volume (BV/TV), trabecular number (Tb.N), bone mineral density (BMD), and bone surface density (BS/TV) in both BCI-treated groups relative to the OVX group.

BCI, as a selective dual-specificity phosphatase 6 (DUSP6) inhibitor, can inhibit tumor growth and macrophage inflammation.^[1].

In vitro, at low-dose of ≤2 μM and ≤4 μM, BCI showed no cytotoxic effects on RAW264.7 cells and BMMs, respectively. And at concentrations of ≤4 μM, BCI had no obvious effect on cell cycle progression or apoptosis in BMMs.^[1]In vitro experiment it shown that treatment with 1 μM BCI enhanced osteoclastogenesis by inhibiting DUSP6. Moreover, BCI increased the levels of osteoclast-related gene expression such as NFATC1, C-fos, ACP5, and DC-STAMP.^[2]In vitro efficacy test it demonstrated that treatment with 4 μm BCI obviously increased the proportion of cells expressing cleaved caspase‐3, 4 μm BCI already elicited extensive cytotoxicity in KELLY and IMR‐32 cells, and only a minority of LAN‐1 and SK‐N‐AS cells remained.^[3]In vitro, with 1 μM BCI did not affect total NCC and NCC surface expression as well as ERK1/2 phosphorylation. Treatment with 5 μM BCI can markedly increase ERK1/2 phosphorylation and decrease total NCC and NCC surface expression.^[5].

In vivo, mice were treated with 10mg/kg BCI intraperitoneally for five consecutive days per week, suppressed AKT activation and prevents tumor formation.^[4]In vivo test it exhibited that treatment with 50, 100, and 200 mg/kg BCI orally in the CPDM animal model obviously increased the number of pNrf2-positive cells in periodontal tissue and mitigated the alveolar bone loss.^[6].

References:
[1] Cai C, et al. BCI Suppresses RANKL-Mediated Osteoclastogenesis and Alleviates Ovariectomy-Induced Bone Loss. Front Pharmacol. 2021 Nov 1;12:772540.
[2] Zhang B, et al. DUSP6 expression is associated with osteoporosis through the regulation of osteoclast differentiation via ERK2/Smad2 signaling. Cell Death Dis. 2021 Sep 2;12(9):825.
[3] Thompson EM, et al. The cytotoxic action of BCI is not dependent on its stated DUSP1 or DUSP6 targets in neuroblastoma cells. FEBS Open Bio. 2022 Jul;12(7):1388-1405.
[4] Duan S, et al. Loss of FBXO31-mediated degradation of DUSP6 dysregulates ERK and PI3K-AKT signaling and promotes prostate tumorigenesis. Cell Rep. 2021 Oct 19;37(3):109870.
[5] Feng X, et al. Aldosterone modulates thiazide-sensitive sodium chloride cotransporter abundance via DUSP6-mediated ERK1/2 signaling pathway. Am J Physiol Renal Physiol. 2015 May 15;308(10):F1119-27.
[6] Zhu C, et al. The therapeutic role of baicalein in combating experimental periodontitis with diabetes via Nrf2 antioxidant signaling pathway. J Periodontal Res. 2020 Jun;55(3):381-391.