Objective To investigate the mechanism of benzyl isothiocyanate(BITC)in the treatment of anaplastic thyroid cancer(ATC).Methods Using network pharmacological analysis,key targets of BITC and ATC were screened,followed...Objective To investigate the mechanism of benzyl isothiocyanate(BITC)in the treatment of anaplastic thyroid cancer(ATC).Methods Using network pharmacological analysis,key targets of BITC and ATC were screened,followed by CO and KEGG enrichment analysis.In order to validate the findings,AutoDock software was used to dock BITC and ATC key targets.BITC was applied to two ATC cell lines(8505C and CAL-62).Flow cytometry was used to analyze cell apoptosis.Autophagy inhibitors hydroxychloroquine sulfate(HCQ)and 3-methyladenine(3MA)were used in combination with BITC.Real-time quantitative PCR was conducted to detect the gene level of LC3B,while Western blotting was utilized to examine the expression of NF-kB,LC3B I,Beclin-1,and Bcl-2.In animal experiments,a mouse tumor model was constructed using CAL-62 cells,treated with intraperitoneal injections of BITC(100 mg/kg)and normal saline respectively,administered every other day for a total of 21 days.Immunoblotting of tumor tissue was performed to detect the expression of LC3B II,Bcl-2,Beclin-1,and NFkB.Results A total of 10 key targets with binding energies≤-4.0 kcal/mol were identified.KECG analysis showed that these genes are mainly involved in NF-kB signaling pathway and apoptosis.BITC inhibited ATC cells with IC50 values of 27.56μmol/L for 8505C and 28.30μmol/L for CAL-62.The expression levels of NF-kB,Beclin-1,and Bcl-2 decreased,while LC3B I and LC3B gene expression increased.Combining 3MA with BITC enhanced cell inhibition LC3B II expression.HCQ increased LC3B II expression without enhancing cell and viability inhibition.In the mouse tumor model,compared to the control group,the treatment group had higher LC3B II and lower Bcl-2,Beclin-1,and NF-kB levels.Conclusion BITC could inhibit the growth of ATC cells in vitro and in vivo,disrupt the autophagy degradation,and inhibit the NF-kB pathway.展开更多
文摘Objective To investigate the mechanism of benzyl isothiocyanate(BITC)in the treatment of anaplastic thyroid cancer(ATC).Methods Using network pharmacological analysis,key targets of BITC and ATC were screened,followed by CO and KEGG enrichment analysis.In order to validate the findings,AutoDock software was used to dock BITC and ATC key targets.BITC was applied to two ATC cell lines(8505C and CAL-62).Flow cytometry was used to analyze cell apoptosis.Autophagy inhibitors hydroxychloroquine sulfate(HCQ)and 3-methyladenine(3MA)were used in combination with BITC.Real-time quantitative PCR was conducted to detect the gene level of LC3B,while Western blotting was utilized to examine the expression of NF-kB,LC3B I,Beclin-1,and Bcl-2.In animal experiments,a mouse tumor model was constructed using CAL-62 cells,treated with intraperitoneal injections of BITC(100 mg/kg)and normal saline respectively,administered every other day for a total of 21 days.Immunoblotting of tumor tissue was performed to detect the expression of LC3B II,Bcl-2,Beclin-1,and NFkB.Results A total of 10 key targets with binding energies≤-4.0 kcal/mol were identified.KECG analysis showed that these genes are mainly involved in NF-kB signaling pathway and apoptosis.BITC inhibited ATC cells with IC50 values of 27.56μmol/L for 8505C and 28.30μmol/L for CAL-62.The expression levels of NF-kB,Beclin-1,and Bcl-2 decreased,while LC3B I and LC3B gene expression increased.Combining 3MA with BITC enhanced cell inhibition LC3B II expression.HCQ increased LC3B II expression without enhancing cell and viability inhibition.In the mouse tumor model,compared to the control group,the treatment group had higher LC3B II and lower Bcl-2,Beclin-1,and NF-kB levels.Conclusion BITC could inhibit the growth of ATC cells in vitro and in vivo,disrupt the autophagy degradation,and inhibit the NF-kB pathway.
