Application of a kind of timosaponin I in the preparation of anti-human ovarian cancer drug
A technology of timosaponin and ovarian cancer, which is applied in the field of application of timosaponin I in the preparation of anti-human ovarian cancer drugs, can solve the problems that the function and mechanism of action have yet to be studied, and timosaponin I has not been discovered yet, achieving a huge The effect of medicinal potential
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Embodiment 1
[0024] The effects of timosaponin I and SHH pathway inhibitor GANT61 on the expression of SKOV3-related proteins in human ovarian cancer cells were determined by Western blotting.
[0025] 1. Experimental materials:
[0026] 1) Human ovarian cancer cell line SKOV3 (ATCC, USA);
[0027] 2) McCoy's5A incomplete medium (KGI, China);
[0028] 3) DMEM high glucose medium powder, GlutaMaxTM glutamine supplement, sodium pyruvate solution and 0.25% EDTA-trypsin (Gibco, USA);
[0029] 4) Fetal bovine serum FBS (Wisent, Canada);
[0030] 5) Penicillin-streptomycin-neomycin mixed antibiotic PSN and BCA protein detection kit (Thermo Fisher, USA);
[0031] 6) Rabbit anti-Gli1 polyclonal antibody (CST Company, USA);
[0032] 7) Rabbit anti-Sufu polyclonal antibody (Abcam, USA);
[0033] 8) Rabbit anti-GAPDH polyclonal antibody (Santa Crutz, USA);
[0034] 9) Horseradish peroxidase-labeled goat anti-mouse or anti-rabbit IgG (Jackson ImmunoResearch, USA);
[0035] 10) GANT61 powder (Se...
Embodiment 2
[0048] The effects of timosaponin I and SHH pathway inhibitor GANT61 on the expression of SKOV3-related mRNA in human ovarian cancer cells were determined by Real-Time PCR.
[0049] 1. Experimental materials:
[0050] 1) Human ovarian cancer cell line SKOV3 (ATCC, USA);
[0051] 2) McCoy's5A incomplete medium (KGI, China);
[0052] 3) DMEM high glucose medium powder, GlutaMaxTM glutamine supplement, sodium pyruvate solution and 0.25% EDTA-trypsin (Gibco, USA);
[0053] 4) Fetal bovine serum FBS (Wisent, Canada);
[0054] 5) Penicillin-streptomycin-neomycin mixed antibiotic PSN (Thermo Fisher Company, USA);
[0055] 6) TRIzol reagent (TaKaRa company, Japan);
[0056] 7) HiScriptⅡQ-RT SuperMix for qPCR (gDNA wiper) reverse transcription kit, AceQ qPCRSYBR Green Master Mix fluorescence quantitative PCR kit (Novizon, China);
[0057] 8) GANT61 powder (Selleck, USA) was dissolved in ethanol with a concentration of 10 mM, diluted with culture medium for experimental use;
[00...
Embodiment 3
[0075] Effects of timosaponin I and SHH pathway inhibitor GANT61 on the proliferation of human ovarian cancer cell SKOV3 by EdU proliferation assay
[0076] 1. Experimental materials:
[0077] 1) Human ovarian cancer cell line SKOV3 (ATCC, USA);
[0078] 2) McCoy's5A incomplete medium (KGI, China);
[0079] 3) DMEM high glucose medium powder, GlutaMaxTM glutamine supplement, sodium pyruvate solution and 0.25% EDTA-trypsin (Gibco, USA);
[0080] 4) Fetal bovine serum FBS (Wisent, Canada);
[0081] 5) Penicillin-streptomycin-neomycin mixed antibiotic PSN (Thermo Fisher Company, USA);
[0082] 6) EdU cell proliferation detection kit (Ribo Biotechnology, China);
[0083] 7) GANT61 powder (Selleck Company, USA) was dissolved in ethanol with a concentration of 10 mM, and diluted with culture medium for experimental use;
[0084] 8) Timosaponin powder (Nanjing University of Traditional Chinese Medicine, China) was dissolved in DMSO with a concentration of 1 mM and diluted with c...
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