Study On Viability Inhibit And Genetic Damage In Cultured Cells Induced By Acrylonitrile

Objective: Acrylonitrile(AN) is an important material used for synthesizing organics.It is hypertoxic and can result in acute and chronic poisoning to exposers. The centralnervous system is the most important target-site. Recently,people pay more attention to itsdelayed effect, including carcinogenicity, teratogenesis and mutagenicity. Reseach showsAN can definitely carcinogenesis to experimental animals. Epidemiological survey findspeople who contact with AN frequently have higher pulmonary carcinoma prevalence. Butup to now, the exact mechanism for carcinogenicity has not been conclusive. To study theeffect of viability inhibit and genetic damage in cultured cells induced by acrylonitrile, wedesigned the whole test. We try to explore the possible mechanism of its carcinogenesis byDNA damage,repair and apoptosis, which may provide some theoretical data forpreventing and treating poisoning induced by AN.Methods: This study includes three parts.In the first part, the viability of V79 cellswas detected by trypan blue stain assay and MTT method and the dose-effect andtime-effect relation could be definited. Secondly, we use Alkli single cell gelelectrophoresis(SCGE) to study the DNA damage level and types of V79 cells which weretreated by different dosage of AN. We also compare the difference of DNA damage andsubsequent repair effect in V79 and SHG4 cells administrated by AN in this way. ConfocalScanning Laser Microscope(CSLM) detected the content of reactive oxygen species(ROS)after the fluorescent probes stained V79 cells which had been treated for two hours by AN.Finally, cell cycle distribute was tested by flow cytometry(FCM) with propidium iodide(PI)stain and apoptosis was observed in V79 cells by CSLM. So we could further comprehendthe genetic mechanism of AN through the whole tests.Conclusion: AN could cause the acute cytotoxicity in V79 cells. The viability ratedecreased in a concentration-dependant and time-dependant mode. But trypan blue stain assay and MTT method had different spectrum of effect on V79 cells. AN could lead tosingle-strand breakage of V79 cells and damage became more severe with increase of AN.Under our experimental condition, DNA cross-linking was not observed in V79 cells. DNAdamage of V79 cells was approximate to that of SHG4 cells when they were exposed to thesame concentration. The damage of both could be repaired when the dosage was low, butnot in high-dose groups even 24 hour later. The content of ROS in V79 cells increasedwith the administrative concentration. We presume that oxidative damage maybe play animportant role in the process of DNA breakage. FCM showed that the proportion of G2phase cells increased in low-dose groups of V79 cells. But cell cycle arrested in G1 phasewith further increase of AN dosage.As for SHG4 cells, cycle was blocked to S and G2phase. We could observe apoptosis of V79 cells treated by AN through CSLM, but necrosishanppened when dose was higher.