中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
10期
2453-2455
,共3页
朱洪旭%匡天涛%戎叶飞%倪晓凌%周文涛%王单松
硃洪旭%劻天濤%戎葉飛%倪曉凌%週文濤%王單鬆
주홍욱%광천도%융협비%예효릉%주문도%왕단송
高迁移率族蛋白B1%RNA,小分子干扰%胰腺癌%化疗
高遷移率族蛋白B1%RNA,小分子榦擾%胰腺癌%化療
고천이솔족단백B1%RNA,소분자간우%이선암%화료
High-mobility group protein B1%RNA,small interfering%Pancreatic cancer%Chemotherapy
目的 观察靶向抑制高迁移率族蛋白B1(HMGB1)的表达对胰腺癌化疗效果的影响.方法 实时荧光定量聚合酶链反应(FQ-PCR)及Western blot检测不同浓度吉西他滨(Gemcitabine)作用胰腺癌细胞PANC-1的HMGB1 mRNA及蛋白表达水平;应用反转录病毒将短发夹RNA(shRNA)转染到PANC-1中,并用Western blot检测HMGB1蛋白的表达;噻唑蓝(MTT)检测PANC-1的存活率及吉西他滨的半数有效浓度(IC50).结果 在1～20 mg/L的作用浓度范围内,随吉西他滨作用浓度的增加,PANC-1中HMGB1 mRNA和蛋白的表达水平均逐渐上调;当吉西他滨作用浓度分别1.0、10.0、100.0 mg/L时,实验组的存活率均低于对照组,分别为(42.9±2.8)％和(57.9±1.5)％、(29.3±2.2)％和(44.4±1.9)％、(6.5±0.6)％和(30.6±1.0)％(P＜0.05);实验组吉西他滨的IC50为(1.41±0.13) mg/L,对照组为(5.99 ±0.17) mg/L,差异有统计学意义(P＜0.01).结论 抑制HMGB1的表达可增加PANC-1对吉西他滨的敏感性.
目的 觀察靶嚮抑製高遷移率族蛋白B1(HMGB1)的錶達對胰腺癌化療效果的影響.方法 實時熒光定量聚閤酶鏈反應(FQ-PCR)及Western blot檢測不同濃度吉西他濱(Gemcitabine)作用胰腺癌細胞PANC-1的HMGB1 mRNA及蛋白錶達水平;應用反轉錄病毒將短髮夾RNA(shRNA)轉染到PANC-1中,併用Western blot檢測HMGB1蛋白的錶達;噻唑藍(MTT)檢測PANC-1的存活率及吉西他濱的半數有效濃度(IC50).結果 在1～20 mg/L的作用濃度範圍內,隨吉西他濱作用濃度的增加,PANC-1中HMGB1 mRNA和蛋白的錶達水平均逐漸上調;噹吉西他濱作用濃度分彆1.0、10.0、100.0 mg/L時,實驗組的存活率均低于對照組,分彆為(42.9±2.8)％和(57.9±1.5)％、(29.3±2.2)％和(44.4±1.9)％、(6.5±0.6)％和(30.6±1.0)％(P＜0.05);實驗組吉西他濱的IC50為(1.41±0.13) mg/L,對照組為(5.99 ±0.17) mg/L,差異有統計學意義(P＜0.01).結論 抑製HMGB1的錶達可增加PANC-1對吉西他濱的敏感性.
목적 관찰파향억제고천이솔족단백B1(HMGB1)적표체대이선암화료효과적영향.방법 실시형광정량취합매련반응(FQ-PCR)급Western blot검측불동농도길서타빈(Gemcitabine)작용이선암세포PANC-1적HMGB1 mRNA급단백표체수평;응용반전록병독장단발협RNA(shRNA)전염도PANC-1중,병용Western blot검측HMGB1단백적표체;새서람(MTT)검측PANC-1적존활솔급길서타빈적반수유효농도(IC50).결과 재1～20 mg/L적작용농도범위내,수길서타빈작용농도적증가,PANC-1중HMGB1 mRNA화단백적표체수평균축점상조;당길서타빈작용농도분별1.0、10.0、100.0 mg/L시,실험조적존활솔균저우대조조,분별위(42.9±2.8)％화(57.9±1.5)％、(29.3±2.2)％화(44.4±1.9)％、(6.5±0.6)％화(30.6±1.0)％(P＜0.05);실험조길서타빈적IC50위(1.41±0.13) mg/L,대조조위(5.99 ±0.17) mg/L,차이유통계학의의(P＜0.01).결론 억제HMGB1적표체가증가PANC-1대길서타빈적민감성.
Objective To investigate the effecof high-mobility group protein B1 ( HMGB1) on the chemosensitivity of pancreaticancer.MethodThe HMGB1 expression levelof PANC-1 treated with differenGemcitabine concentrationwere measured by both the Western blotting and real-time fluorescenquantitative polymerase chain reaction (FQ-PCR).The recombinancontaining HMGB1 shorhairpin RN(shRNA) watransfected into PANC-1 cellby retroviruand the HMGB1 protein level wadetected by Western blotting aftetransfection.Cell viability and half inhabitation concentration (IC50) of Gemcitabine were evaluated by methyl thiazol tetrazolium (MTT) assay.ResultGemcitabine enhanced the expression of HMGB1 in pancreaticancecell line PANC-1.Thieffecwadose dependenwhile the Gemcitabine concentration ranged from 1 mg/L to 20 mg/L.HMGB1 shRNtransfection led to significandecreased HMGB1 in PANC-1.Afteexposed to 1.0, 10.0 and 100.0 mg/L Gemcitabine fo48 hours, the cell viability of interference group wa(42.9 ± 2.8) ％, (29.3 ± 2.2) ％ and (6.5 ± 0.6) ％ respectively, and the control group had highecell viability which wa(57.9 ± 1.5) ％, (44.4 ± 1.9) ％ and (30.6 ± 1.0)％.Moreover, Gemcitabine IC50 in the interference group ilowethan the control group: (1.41 ± 0.13) mg/L vs.(5.99 ± 0.17) mg/L (P ＜ 0.01).Conclusion Suppression of HMGB1 increasesensitivity to Gemcitabine in PANC-1 cells.
