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Inhibition of non-small cell lung cancer (NSCLC) growth by a novel small molecular inhibitor of EGFR.

Li J, Deng H, Hu M, Fang Y, Vaughn A, Cai X, Xu L, Wan W, Li Z, Chen S, Yang X, Wu S, Xiao J - Oncotarget (2015)

Bottom Line: Using in vitro EGFR kinase activity system, we identified a novel small molecule, WB-308, as an inhibitor of EGFR.WB-308 impaired the phosphorylation of EGFR, AKT, and ERK1/2 protein.WB-308 was less cytotoxic than Gefitinib.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Oncology, Changzheng Hospital, The Second Military Medical University, Shanghai 200003, China.

ABSTRACT
The epidermal growth factor receptor (EGFR) is a therapeutic target (oncotarget) in NSCLC. Using in vitro EGFR kinase activity system, we identified a novel small molecule, WB-308, as an inhibitor of EGFR. WB-308 decreased NSCLC cell proliferation and colony formation, by causing G2/M arrest and apoptosis. Furthermore, WB-308 inhibited the engraft tumor growths in two animal models in vivo (lung orthotopic transplantation model and patient-derived engraft mouse model). WB-308 impaired the phosphorylation of EGFR, AKT, and ERK1/2 protein. WB-308 was less cytotoxic than Gefitinib. Our study suggests that WB-308 is a novel EGFR-TKI and may be considered to substitute for Gefitinib in clinical therapy for NSCLC.

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Related in: MedlinePlus

WB-308 inhibits colony formation of NSCLC cells(A) NSCLC cell line SPC-A1 was seeded in 6-well plates for 7 days after the treatment of WB-308 in according concentrations and fixed with 4% paraformaldehyde, and stained with 0.2% crystal violet. (B) Statistic results of 2-D colony formation. Columns, mean; bars, SE (n = 3; t-test, P < 0.05). (C) NSCLC cell line SPC-A1 was subjected to the 3-D colony formation assay described in Materials and Methods and the result were photographed. (D, E) Statistic results of 3-D colony formation. Columns, mean; bars, SE (n = 3; t test, P < 0.05).
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Figure 4: WB-308 inhibits colony formation of NSCLC cells(A) NSCLC cell line SPC-A1 was seeded in 6-well plates for 7 days after the treatment of WB-308 in according concentrations and fixed with 4% paraformaldehyde, and stained with 0.2% crystal violet. (B) Statistic results of 2-D colony formation. Columns, mean; bars, SE (n = 3; t-test, P < 0.05). (C) NSCLC cell line SPC-A1 was subjected to the 3-D colony formation assay described in Materials and Methods and the result were photographed. (D, E) Statistic results of 3-D colony formation. Columns, mean; bars, SE (n = 3; t test, P < 0.05).

Mentions: Two kinds of colony formation assays were performed to test the inhibition effect of WB-308 to NSCLC cell proliferation. One was the 2-D colony formation assay and the other was the 3-D colony formation assay. As Figure 4 shows, WB-308 inhibited colony formation of NSCLC cells in a concentration-dependent manner, and showed a very significant difference compared to the control group when at 10 μM. In Figure 4A, after being seeded in 6 cm dishes and colony formatted for 2 weeks, SPC-A1 cells displayed a decreased number of colonies with the increase of WB-308 concentration (Figure 4A, the upper panel). The bottom panel of Figure 4A illustrates the enlarged view of the respective colony. Figure 4B, 4D and 4E show the statistic results of each colony formation assay, according to the diagram of colony numbers and colony diameters. The 3-D colony formation morphology results are demonstrated in Figure 4C. From Figure 4 we can summarize that WB-308 inhibits colony formation of NSCLC cells.


Inhibition of non-small cell lung cancer (NSCLC) growth by a novel small molecular inhibitor of EGFR.

Li J, Deng H, Hu M, Fang Y, Vaughn A, Cai X, Xu L, Wan W, Li Z, Chen S, Yang X, Wu S, Xiao J - Oncotarget (2015)

WB-308 inhibits colony formation of NSCLC cells(A) NSCLC cell line SPC-A1 was seeded in 6-well plates for 7 days after the treatment of WB-308 in according concentrations and fixed with 4% paraformaldehyde, and stained with 0.2% crystal violet. (B) Statistic results of 2-D colony formation. Columns, mean; bars, SE (n = 3; t-test, P < 0.05). (C) NSCLC cell line SPC-A1 was subjected to the 3-D colony formation assay described in Materials and Methods and the result were photographed. (D, E) Statistic results of 3-D colony formation. Columns, mean; bars, SE (n = 3; t test, P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4466647&req=5

Figure 4: WB-308 inhibits colony formation of NSCLC cells(A) NSCLC cell line SPC-A1 was seeded in 6-well plates for 7 days after the treatment of WB-308 in according concentrations and fixed with 4% paraformaldehyde, and stained with 0.2% crystal violet. (B) Statistic results of 2-D colony formation. Columns, mean; bars, SE (n = 3; t-test, P < 0.05). (C) NSCLC cell line SPC-A1 was subjected to the 3-D colony formation assay described in Materials and Methods and the result were photographed. (D, E) Statistic results of 3-D colony formation. Columns, mean; bars, SE (n = 3; t test, P < 0.05).
Mentions: Two kinds of colony formation assays were performed to test the inhibition effect of WB-308 to NSCLC cell proliferation. One was the 2-D colony formation assay and the other was the 3-D colony formation assay. As Figure 4 shows, WB-308 inhibited colony formation of NSCLC cells in a concentration-dependent manner, and showed a very significant difference compared to the control group when at 10 μM. In Figure 4A, after being seeded in 6 cm dishes and colony formatted for 2 weeks, SPC-A1 cells displayed a decreased number of colonies with the increase of WB-308 concentration (Figure 4A, the upper panel). The bottom panel of Figure 4A illustrates the enlarged view of the respective colony. Figure 4B, 4D and 4E show the statistic results of each colony formation assay, according to the diagram of colony numbers and colony diameters. The 3-D colony formation morphology results are demonstrated in Figure 4C. From Figure 4 we can summarize that WB-308 inhibits colony formation of NSCLC cells.

Bottom Line: Using in vitro EGFR kinase activity system, we identified a novel small molecule, WB-308, as an inhibitor of EGFR.WB-308 impaired the phosphorylation of EGFR, AKT, and ERK1/2 protein.WB-308 was less cytotoxic than Gefitinib.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Oncology, Changzheng Hospital, The Second Military Medical University, Shanghai 200003, China.

ABSTRACT
The epidermal growth factor receptor (EGFR) is a therapeutic target (oncotarget) in NSCLC. Using in vitro EGFR kinase activity system, we identified a novel small molecule, WB-308, as an inhibitor of EGFR. WB-308 decreased NSCLC cell proliferation and colony formation, by causing G2/M arrest and apoptosis. Furthermore, WB-308 inhibited the engraft tumor growths in two animal models in vivo (lung orthotopic transplantation model and patient-derived engraft mouse model). WB-308 impaired the phosphorylation of EGFR, AKT, and ERK1/2 protein. WB-308 was less cytotoxic than Gefitinib. Our study suggests that WB-308 is a novel EGFR-TKI and may be considered to substitute for Gefitinib in clinical therapy for NSCLC.

Show MeSH
Related in: MedlinePlus