The Study Of The Inhibitory Mechanism Of Disulfiram Metabolic Chelates On Non-small Cell Lung Cancer Cells

Disulfiram(DSF)is a drug used to treat alcohol abuse with a history of nearly 70 years,Its safety has been widely certified.After entering the blood,the metabolite DDC(diethyldithiocarbamic acid)of DSF can quickly form DDC-Cu(diethyldithiocarbamate–copper)chelate with divalent copper ions in the blood,The chelate has a strong and specific effect of inhibiting tumor cells.In this experiment,DDC-Cu treatment was performed on different non-small cell lung cancer(NSCLC)cells and the cell survival,apoptosis,cell cycle,cell autophagy related indicators were detected.It was found that DDC-Cu can significantly inhibit the proliferation of NSCLC cells,induce cell death,and lead to changes in molecular indicators related to autophagy and apoptosis.furthermore,the function inhibition and cell research model of DDC-Cu chelate on NSCLC cells in vitro was verified and established.We Used stable isotope labeling by amino acids in cell culture(SILAC)to identify differential proteins in A549 cells after DDC-Cu treatment.Through repetition and screening,72 upregulated proteins and 43 down-regulated proteins were identified.Combining bioinformatics and research literature,we found: NOP53 ribosome biogenesis factor(NOP53),ribonucleotide reductase regulatory subunit M2(RRM2),serine peptidase inhibitor kunitz type 2(SPINT2),minichromosome maintenance complex Component 2(MCM2)related changes may be related to tumor suppressor function.Further experiments we found that has little effect on the DDC-Cu sensitivity of NSCLC cells after using small interfering RNA(si RNA)to downregulate NOP53,NOP53 may be the result of drug treatment and does not affect participation in related functional regulation;In contrast,when we downregulated RRM2 using si RNA interference,we found that it can inhibit NSCLC cell proliferation,promote apoptosis and autophagy,block the cell cycle,and enhance cell sensitivity to DDC-Cu drugs.At the same time,the down-regulation of RRM2 protein occurred in the early stage of DDC-Cu drug-treated cells,earlier than the appearance of the apoptotic molecular marker PAPR1 spliceosome,combined with RRM2 protein function,suggesting that RRM2 may be involved in DDC-Cu for NSCLC cells The tumor suppressor function regulation,or one of the key factors.In summary,by establishing and verifying the DDC-Cu drug treatment NSCLC cell model,we used the proteomic SILAC labeling technology and bioinformatics analysis,we discovered and verified that the ability of DDC-Cu to inhibit tumors may be related to RRM2 protein,and the upstream and downstream regulatory proteins related to RRM2 and key signaling molecules and functional signaling pathways need to be further verified.