ObjectiveTo explore the active ingredients， therapeutic targets， and relative signaling pathways of Tripterygium wilfordii in the treatment of triple negative breast cancer （TNBC） based on network pharmacology， and to verify the mechanism through in vitro cell model. MethodThe active ingredients of T. wilfordii were screened from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）. The targets of TNBC were obtained from DisGeNET and GeneCards. Venny was used to identify the potential therapeutic targets of T. wilfordii against TNBC. Protein-protein interaction （PPI） network was constructed with String database. Gene ontology （GO） annotation and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment were carried out with DAVID to predict the mechanisms of potential targets. The molecular docking between triptolide and key targets were performed with AutoDock Vina. The effect of triptolide （0， 5， 10， 20， 30， 40， 50， 60， 80 nmol·L^-1） on the proliferation of MDA-MB-231 cells was determined through methyl thiazolyl tetrazolium （MTT） assay. The effect of triptolide （0， 12.5， 25， 50 nmol·L^-1） on the apoptosis of MDA-MB-231 cells was detected with Hoechst 33342 staining. Western blot was performed to detect the effect of triptolide （0， 25， 50 nmol·L^-1） on the expression levels of key targets. ResultT. wilfordii had 23 active ingredients related to 55 potential targets of TNBC. GO and KEGG enrichment revealed that the potential targets were associated with 103 biological processes， 15 cellular components， and 35 molecular functions， and were involved in 140 signaling pathways including atherosclerosis and apoptosis. The results of molecular docking demonstrated that triptolide could bind with the targets including threonine kinase 1 （Akt1）， vascular endothelial growth factor A （VEGFA）， cellular tumor antigen p53 （p53）， transcription factor AP-1 （JUN）， signal transducer and activator of transcription 3 （STAT3）， tumor necrosis factor （TNF）， mitogen-activated protein kinase 8 （MAPK8）， prostaglandin G/H synthase 2 （PTGS2）， and Caspase-3. According to the results of MTT assay， triptolide （20， 30， 40， 50， 60， 80 nmol·L^-1） inhibited the proliferation of MDA-MB-231 cells compared with blank control （P<0.05， P<0.01）. Hoechst 33342 staining showed that triptolide （12， 25， 50 nmol·L^-1） induced the apoptosis of MDA-MB-231 cells compared with black control （P<0.05， P<0.01）. Western blot showcased that 50 nmol·L^-1 triptolide down-regulated the relative expression levels of p-Akt， TNF-α， and VEGFA， while 25 and 50 nmol·L^-1 triptolide up-regulated the relative expression level of p53 in a dose-dependent manner compared with the blank control （P<0.05， P<0.01）. ConclusionT. wilfordii has multiple ingredients， targets， and pathways in the treatment of TNBC. It may regulate p53， VEGFA， TNF-α and other key targets to induce cell apoptosis and suppress angiogenesis and inflammatory response， which provides a scientific basis for the further investigation and clinical application of T. wilfordii.

Objective To investigate the inhibit proliferation effect of Gecko crude peptide (GCP)on human esophageal carcinoma KYSE450 cells in vitro and to explore its mechanism.Methods KYSE450 cells were put into plates in its logarithmic phase,and they were treated with different concentration (0.10,0.15,0.20,0.25,0.30,0.35,0.4,0.45,0.50,0.55,0.6 mg · mL-1) of GCP for 24,48,72 h,and then detected corresponding indicators with respective methods.The viability of KYSE450 cells was detected with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT').The concentration of blank group(GCP:0),low-dose,middle-dose and high-dose experimental groups(GCP:0.10,0.15,0.22 mg · mL-1)groups were separately,according to the results of MTT.Adriamycin was chosen as the control drug,which concentration was 10 μg · mL-1.After treatment with GCP,the expression of B-cell lymphoma-2(Bcl-2),Bel-2 associated X protein (Bax),cleaved cysteinyl aspartate specific proteinase-3 (Caspase-3),cleaved cysteinyl aspartate specific proteinase-9 (Caspase-9) proteins in KYSE450 cells were observed by Western blot.Results The GCP significantly inhibited proliferation of KYSF450 cells in a dose-and time-dependent manners.After treatment of GCP for 24,48,72 h,the 50％ inhibitory dose (IC50) values were 0.32,0.28,0.24 mg · mL-1,respectively.The Bax and Bcl-2 grayscale average ratio in blank group,low-dose,middle-dose and high-dose experimental groups,control group were (23.83 ± 0.07)％,(51.08 ± 0.78)％,(148.26 ± 6.19) ％,(329.33 ± 4.44) ％,(115.84 ± 0.26) ％.The Caspase-9 and glyceraldehyde phosphate dehydrogenase (GAPDH) grayscale average ratio in above five groups were (14.81 ± 1.05) ％,(20.91 ± 1.24) ％,(39.61 ± 1.89) ％,(43.68 ± 1.13) ％,(39.82 ± 0.69) ％.The Caspase-3 and GAPDH grayscale average ratio in above five groups were (6.37 ±0.51)％,(10.97 ± 0.50)％,(13.44 ± 0.49)％,(17.03 ±0.49)％,(8.63 ± 0.42) ％.Compared with blank group,the differences in drugs groups were significantly (P ＜ 0.05,P ＜ 0.01) Conclusion GCP can inhibit proliferation and induce apoptosis of KYSE450 cells,which may be associated with a Caspase-dependent manner in KYSE450 cells.