Screening And Verification Of Anti-tumor Target Of Novel Monoterpenoid Alkaloid TBCN

Objective:The anti-tumor target of the novel monoterpenoid alkaloid TBCN was screened and verified.Methods:1?Antiproliferative activity of compound TBCN against human melanoma cell line A375,human non-small cell lung cancer cell line A549,human liver cancer cell line HepG2,human ovarian cancer cell line SKOV3 and human colon cancer cell line HCT116 by MTT assay.2?According to the results of the previous test,A549 cells were selected and PCR arrays were used to screen the possible targets of compound TBCN.3?According to the results of PCR array,the effects of compound TBCN on the transcription of BIRC3,ATM,CCND3,CCNE1,APC,KRAS,ITGA7,MAP2K1,JUN,CCND2,CDK6,NCOA1 and other genes were verified by RQ-PCR.Results:1?The results of MTT array showed that among the 5 tumor cells tested,the compound TBCN showed significant activity with an average half-inhibitory concentration(IC50)of less than 3 ?M.In addition to the activity of compound TBCN on SKOV3 cells was lower than its positive control paclitaxel,the activity of compound TBCN was higher than the corresponding positive control for A375,A549,HepG2 and HCT116 cells,especially in A375 and A549 cells.2?The results of PCR array screening showed that the compound TBCN significantly up-regulated the transcription of BIRC3,ATM,CCND3,CCNE1,APC,KRAS genes and in A549 cells,and down-regulated the transcription of MAP2K1,JUN,CCND2,CDK6 and NR4A3 genes.This suggests that the inhibition of A549 cells by the compound TBCN may be related to the signal transduction pathway associated with the above genes with significant changes in transcription levels.3?The results of RQ-PCR showed that the compound TBCN up-regulated the transcription of BIRC3,ATM,CCNE1,APC and ITGA7 genes;down-regulated the transcription of CCND2 and CCND3 genes;the transcription of CDK6,MAP2K1,JUN genes was down-regulated,but not significant;the transcription of NCOA1 gene has no significant effect;the effect on KRAS gene transcription is uncertain.Conclusion:The compound TBCN inhibits the proliferation of A549 cells involved in a variety of tumor-associated regulatory genes and multiple signal transduction pathways.The interrelationship and mechanism of action between them requires a large number of in-depth studies to answer.However,many of the results of this study have laid the foundation and basis for further research.