中华胃肠外科杂志
中華胃腸外科雜誌
중화위장외과잡지
CHINESE JOURNAL OF GASTROINTESTINAL SURGERY
2013年
6期
578-582
,共5页
曹明争%毛伟征%马贵亮%李杨
曹明爭%毛偉徵%馬貴亮%李楊
조명쟁%모위정%마귀량%리양
胃肿瘤%肿瘤坏死因子-α%核转录因子-κb%吡咯烷二硫代氨基甲酸盐%凋亡
胃腫瘤%腫瘤壞死因子-α%覈轉錄因子-κb%吡咯烷二硫代氨基甲痠鹽%凋亡
위종류%종류배사인자-α%핵전록인자-κb%필각완이류대안기갑산염%조망
Stomach neoplasms%Tumor necrosis factor%Nuclear factor-κb%Pyrrolidine dithio-carbamate%Apoptosis
目的 研究核转录因子-κb(NF-κb)抑制剂吡咯烷二硫代氨基甲酸盐(PDTC)对肿瘤坏死因子α(TNF-α)诱导的人胃癌细胞株SGC-7901生长抑制及凋亡的影响,并探讨其作用机制.方法 应用噻唑蓝(MTT)法检测不同浓度的PDTC和TNF-α以及两者联合应用对SGC-7901细胞增殖的抑制率；采用Hoechst检测SGC-7901细胞凋亡情况；Western blot检测SGC-7901细胞survivin和caspase-3蛋白的表达.结果 PDTC在15、30、60和100 μmol/L浓度时,对SGC-7901的细胞生长抑制率分别为(12.14±0.91)％、(20.00±1.11)％、(37.63±1.01)％和(41.46±1.07)％,均可抑制细胞增殖(P＜0.01).TNF-α为25 mg/L时,对SGC-7901细胞的生长抑制率为(2.38±0.67)％,与对照组(1.50±0.81)％相比,差异无统计学意义(F=28.28,P＞0.05)；而在50、100和150 mg/L浓度时,对SGC-7901细胞的生长抑制率分别为(4.53±0.85)％、(4.43±0.70)％和(4.74±1.07)％,与对照组相比,差异有统计学意义(P＜0.05).PDTC 15 μmol/L分别与25、50、100和150 mg/L的TNF-α联合应用时,对SGC-7901的细胞生长抑制率则分别为(18.94±1.10)％、(30.23±0.89)％、(41.55±0.94)％和(53.34±0.98)％,与单用TNF-α或单用15 μmol/L PDTC比较,细胞生长抑制率增加(P＜0.01).Hoechst检测结果显示,TNF-α 100 mg/L组、PDTC 15 μmol/L组及两者联合应用组细胞凋亡率均显著增加(P＜0.01),且联合用药组细胞凋亡率增高最为显著(P＜0.01).PDTC(15μmol/L)与TNF-α(100 mg/L)联合用药与单用TNF-α(100 mg/L)比较,细胞survivin蛋白表达明显降低(P＜0.01),与单用PDTC(15 μmol/L)比较,差异无统计学意义(P＞0.05)；但caspase-3蛋白的表达联合用药组较两者分别单用时显著增加(P＜0.01).结论 PDTC可增强TNF-α对人SGC-7901细胞的促凋亡作用,其机制可能与PDTC阻断TNF-α诱导的NF-κb活性、下调survivin表达并最终上调凋亡蛋白caspase-3的表达有关.
目的 研究覈轉錄因子-κb(NF-κb)抑製劑吡咯烷二硫代氨基甲痠鹽(PDTC)對腫瘤壞死因子α(TNF-α)誘導的人胃癌細胞株SGC-7901生長抑製及凋亡的影響,併探討其作用機製.方法 應用噻唑藍(MTT)法檢測不同濃度的PDTC和TNF-α以及兩者聯閤應用對SGC-7901細胞增殖的抑製率；採用Hoechst檢測SGC-7901細胞凋亡情況；Western blot檢測SGC-7901細胞survivin和caspase-3蛋白的錶達.結果 PDTC在15、30、60和100 μmol/L濃度時,對SGC-7901的細胞生長抑製率分彆為(12.14±0.91)％、(20.00±1.11)％、(37.63±1.01)％和(41.46±1.07)％,均可抑製細胞增殖(P＜0.01).TNF-α為25 mg/L時,對SGC-7901細胞的生長抑製率為(2.38±0.67)％,與對照組(1.50±0.81)％相比,差異無統計學意義(F=28.28,P＞0.05)；而在50、100和150 mg/L濃度時,對SGC-7901細胞的生長抑製率分彆為(4.53±0.85)％、(4.43±0.70)％和(4.74±1.07)％,與對照組相比,差異有統計學意義(P＜0.05).PDTC 15 μmol/L分彆與25、50、100和150 mg/L的TNF-α聯閤應用時,對SGC-7901的細胞生長抑製率則分彆為(18.94±1.10)％、(30.23±0.89)％、(41.55±0.94)％和(53.34±0.98)％,與單用TNF-α或單用15 μmol/L PDTC比較,細胞生長抑製率增加(P＜0.01).Hoechst檢測結果顯示,TNF-α 100 mg/L組、PDTC 15 μmol/L組及兩者聯閤應用組細胞凋亡率均顯著增加(P＜0.01),且聯閤用藥組細胞凋亡率增高最為顯著(P＜0.01).PDTC(15μmol/L)與TNF-α(100 mg/L)聯閤用藥與單用TNF-α(100 mg/L)比較,細胞survivin蛋白錶達明顯降低(P＜0.01),與單用PDTC(15 μmol/L)比較,差異無統計學意義(P＞0.05)；但caspase-3蛋白的錶達聯閤用藥組較兩者分彆單用時顯著增加(P＜0.01).結論 PDTC可增彊TNF-α對人SGC-7901細胞的促凋亡作用,其機製可能與PDTC阻斷TNF-α誘導的NF-κb活性、下調survivin錶達併最終上調凋亡蛋白caspase-3的錶達有關.
목적 연구핵전록인자-κb(NF-κb)억제제필각완이류대안기갑산염(PDTC)대종류배사인자α(TNF-α)유도적인위암세포주SGC-7901생장억제급조망적영향,병탐토기작용궤제.방법 응용새서람(MTT)법검측불동농도적PDTC화TNF-α이급량자연합응용대SGC-7901세포증식적억제솔；채용Hoechst검측SGC-7901세포조망정황；Western blot검측SGC-7901세포survivin화caspase-3단백적표체.결과 PDTC재15、30、60화100 μmol/L농도시,대SGC-7901적세포생장억제솔분별위(12.14±0.91)％、(20.00±1.11)％、(37.63±1.01)％화(41.46±1.07)％,균가억제세포증식(P＜0.01).TNF-α위25 mg/L시,대SGC-7901세포적생장억제솔위(2.38±0.67)％,여대조조(1.50±0.81)％상비,차이무통계학의의(F=28.28,P＞0.05)；이재50、100화150 mg/L농도시,대SGC-7901세포적생장억제솔분별위(4.53±0.85)％、(4.43±0.70)％화(4.74±1.07)％,여대조조상비,차이유통계학의의(P＜0.05).PDTC 15 μmol/L분별여25、50、100화150 mg/L적TNF-α연합응용시,대SGC-7901적세포생장억제솔칙분별위(18.94±1.10)％、(30.23±0.89)％、(41.55±0.94)％화(53.34±0.98)％,여단용TNF-α혹단용15 μmol/L PDTC비교,세포생장억제솔증가(P＜0.01).Hoechst검측결과현시,TNF-α 100 mg/L조、PDTC 15 μmol/L조급량자연합응용조세포조망솔균현저증가(P＜0.01),차연합용약조세포조망솔증고최위현저(P＜0.01).PDTC(15μmol/L)여TNF-α(100 mg/L)연합용약여단용TNF-α(100 mg/L)비교,세포survivin단백표체명현강저(P＜0.01),여단용PDTC(15 μmol/L)비교,차이무통계학의의(P＞0.05)；단caspase-3단백적표체연합용약조교량자분별단용시현저증가(P＜0.01).결론 PDTC가증강TNF-α대인SGC-7901세포적촉조망작용,기궤제가능여PDTC조단TNF-α유도적NF-κb활성、하조survivin표체병최종상조조망단백caspase-3적표체유관.
Objective To investigate the effect of PDTC (inhibitor of NF-κb) on apoptosis of human gastric cancer cell line SGC-7901 induced by tumor necrosis factor α (TNF-α) and explore the related mechanisms.Methods After the treatment with different concentrations of PDTC,TNF-α or PDTC combined with TNF-α on gastric cancer cell line SGC-7901,the growth inhibition of SGC-7901 was measured by MTT assay.Hoechst was used to assess SGC-7901 cell apoptosis.The protein expressions of survivin and caspase-3 were detected by Western blot assay.Results The growth inhibition rate of SGC-7901 induced by PDTC (15,30,60,100 μmol/L) was (12.14Π0.91)％,(20.00±1.11)％,(37.63±1.01)％ and (41.46±1.07)％.Different concentrations of PDTC all inhibited the growth of SGC-7901 significantly (all P＜0.01),The growth inhibition rate of SGC-7901 induced by 25 mg/L TNF-α was (2.38±0.67)％,which could not significantly inhibit the growth of SGC-7901 [control (1.50±0.81)％],while TNF-α of 50,100,150 mg/L could inhibit the growth of SGC-7901 significantly [(4.53±0.85)％,(4.43±0.70)％ and(4.74±1.07)％,all P＜0.05].PDTC (15 μmol/L) combined with TNF-α (25,50,100,150 mg/L) significantly increased the cell growth inhibition rate compared with TNF-α alone or PDTC 15 μmol/L alone (all P＜0.01).Hoechst assay showed that 100 mg/L TNF-α,15 μmol/L PDTC and combination of above two all induced cell apoptosis (P＜0.01),and the combination group had significantly higher percentage of cell apoptosis(P＜0.01).Survivin protein was significantly down-regulated in combination group as compared with single TNF-α (100 mg/L) group,but was not significant down-regulated as compared with single PDTC (15 μmol/L) group.Caspase-3 protein expression was significantly increased in combination group as compared with other two groups.Conclusion PDTC can enhance the cell apoptosis induced by TNF-α,which may be associated with the blocking of TNF-α-activated NF-κB signaling pathway by PDTC,the down-regulation of survivin expression,and up-regulation of caspase-3 expression.
