Medicine composition for treating neuroblastoma
A neuroblastoma and composition technology, applied in the field of pharmaceutical compositions for the treatment of neuroblastoma, can solve problems such as complete remission and low
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Embodiment 1
[0100] Embodiment 1. Effect and molecular mechanism of NVP-BEZ235 on neuroblastoma cells
[0101] 1. CCK-8 method to detect the effect of NVP-BEZ235 on the proliferation of neuroblastoma
[0102] In order to investigate the effect and molecular mechanism of the PI3K / mTOR dual inhibitor NVP-BEZ235 on neuroblastoma cells, the inventors first treated the neuroblastoma cell line SHSY-5Y with NVP-BEZ235 at concentrations of 200, 500, and 1000 nM and SK-N-MC, the treatment time was 12h or 24h, and the effect of NVP-BEZ235 on the proliferation of neuroblastoma was detected by CCK-8 method.
[0103] It was found that NVP-BEZ235 had a significant inhibitory effect on the proliferation of neuroblastoma cell lines in a time- and dose-dependent manner, as shown in figure 1 A and figure 1 Shown in B.
[0104] 2. Detection of the inhibitory effect of NVP-BEZ235 on neuroblastoma cell lines by immunoblotting
[0105] Furthermore, the present inventors detected the effects of NVP-BEZ235 on...
Embodiment 2
[0108] Embodiment 2, the influence of NVP-BEZ235 on cell cycle
[0109] 1. Analysis of the effect of NVP-BEZ235 on the cell cycle of neuroblastoma cells by flow cytometry
[0110] In order to further explore the mechanism of the inhibitory effect of NVP-BEZ235 on the proliferation of neuroblastoma cell lines, flow cytometry was used to detect the effect of NVP-BEZ235 on the cell cycle.
[0111] The result is as figure 2 A and figure 2 As shown in B, after neuroblastoma cell lines SHSY-5Y and SK-N-MC were treated with 500 and 1000 nM NVP-BEZ235 for 24 hours, the number of cells in S phase and G2 / M phase was significantly down-regulated, but no sub-G1 population appeared.
[0112] 2. Western blot analysis of the protein levels of cyclinD1 and cyclinE in NVP-BEZ235-treated neuroblasts
[0113] Further, in order to clarify the effect of NVP-BEZ235 on the G0 / G1 phase arrest of neuroblasts, the inventors analyzed the protein levels of cyclinD1 and cyclinE in neuroblasts treated...
Embodiment 3
[0115] Example 3, NVP-BEZ235 and Rubescensine A have a synergistic effect
[0116] 1. CCK-8 method to detect the effect of NVP-BEZ235 on the proliferation of neuroblastoma
[0117] SHSY-5Y (A) and SK-N-MC (B) cells were treated with different concentrations of Rubescensine A for 24 hours, and CCK8 was used to detect the cell proliferation level.
[0118] The results showed that Rubescensine A had a significant inhibitory effect on the proliferation of neuroblastoma cell lines, and it was time- and dose-dependent, as shown in Figure 4 A and Figure 4 Shown in B.
[0119] 2. Determination of the synergistic effect of NVP-BEZ235 and Rubescensin A
[0120] The inhibitory effects of different formulations of NVP-BEZ235 and Rubescensine A on the proliferation of neuroblastoma cells SH-SY-5Y and SK-N-MC were detected. Add NVP-BEZ235 and Oridonin to the 96-well plate of cultured neuroblastoma cells SH-SY-5Y and SK-N-MC according to the addition amount listed in Table 1, treat for...
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