Based On Caenorhabditis Elegans Model To Explore The Anti-tumor Mechanism Of Apoptotic Small Molecules

Small-molecule anti-tumor drugs occupy a dominant position in the new anti-tumor drugs,and have shown a blowout development in recent years.Abnormalities related to signal perception,integration and execution of apoptosis may lead to tumors.Therefore,designing and screening small molecule drugs targeting key molecules of apoptosis signals is the most effective way to fight cancer.N-(3-oxododecanoyl)-L-homoserine lactone(N-(3-oxododecanoyl)-L-homoserine lactone,hereinafter referred to as C12)and ABT-737 are two pro-apoptotic small molecule that are closely related to Bcl-2 family proteins.Existing data indicate that the two drugs can promote tumor cell apoptosis signaling at the cellular level.At present,the effects of C12 and ABT-737 on cell apoptosis and their anti-tumor mechanisms at the living level are not clear.As a model animal,Caenorhabditis elegans is conserved with human genes,and has developed an experimental model of nematodes for tumor research.Therefore,this article uses Caenorhabditis elegans as a multi-cell living animal model to deeply study the effects of C12 and ABT-737 on apoptosis at the living level,and discuss in detail the effect and mechanism on C.elegans tumor model.Here are the results:1.After treating C.elegans with different concentration gradients of C12,it was found that all the selected concentrations showed the effect of promoting nematode gonad cell apoptosis.We further explored the molecular signaling mechanism of C12 in the induction of nematode gonad cell apoptosis.Using gene mutation strains and real-time quantitative PCR technology,we found that core apoptotic pathway genes egl-1,ced-4,ced-3,DNA damage checkpoints hus-1 and clk-2,p53 homologous protein cep-1 are involved C12 induced nematode germ cell apoptosis;using H2DCF-DA probe and fluorescent strain nematode to detect reactive oxygen species(ROS)levels,it was found that C12 induced nematode germ cell apoptosis by causing oxidative stress.In addition,N-acetyl-L-cysteine(NAC),a ROS scavenger,can inhibit the increased expression of C12-induced DNA damage response genes hus-1 and clk-2 and cep-1,indicating that C12 regulates DNA damage through ROS to induce nematode germ cell apoptosis.2.We further explored the effect and mechanism of C12 on nematode vulvar tumors.Using the nematode strain let-60(n1046)with over-activated RAS proto-oncogene to verify the antitumor activity of C12,we found that C12 can effectively inhibit the parental nematode pyloric tumor and restore the mutant phenotype to the wild phenotype,while the progeny nematode genital tumors are not inhibited and still have a mutant phenotype;using H2DCF-DA probes and multiple vaginal door inhibition experiments,NAC can antagonize the nematode tumor suppressive effect induced by C12 due to elevated ROS levels.In addition,real-time quantitative PCR found that C12 inhibited nematode tumor growth by blocking RAS/MAPK signaling.3.After treating wild-type N2 nematodes with different concentrations of ABT-737,we found that ABT-737 induced apoptosis of C.elegans gonad cells in a dose-dependent manner.We further explored the molecular signaling mechanism of ABT-737 in inducing nematode gonad cell apoptosis.Fluorescent nematode strain CF1553(SOD3::GFP)was used to detect ROS levels in nematodes,and it was found that ABT-737 may cause apoptosis by regulating the production of free radicals in the nematode;using gene mutation strains and ABT-73 7 treatment,it was found that DNA damage response genes hus-1,cep-1 and its target gene egl-1 and MAPK pathway genes sek-1,mek-1 and jnk-1 are indispensable for ABT-737-induced germ cell apoptosis.The DNA damage response pathway and MAPK pathway are involved in regulating ABT-737-induced nematode germ cell apoptosis.In this study,the molecular mechanism of C12 and ABT-737 inducing apoptosis and inhibiting tumor at the level of C.elegans was investigated.The results suggest that C12 and ABT-737 can induce apoptosis of C.elegans germ cells,activate oxidative stress and regulate the DNA damage response pathway and MAPK pathway,and C12 can also inhibit cryptic tumors that are excessively activated by C.elegans Ras/MAPK.The research results have some guiding significance for the design and screening of small molecule drugs targeting key molecules of apoptotic signal and provide a good reference value for the study of small molecule anti-tumor drugs in vivo.