中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2014年
4期
238-242
,共5页
胰腺肿瘤%硼替佐米%细胞系,肿瘤%细胞增殖%细胞凋亡
胰腺腫瘤%硼替佐米%細胞繫,腫瘤%細胞增殖%細胞凋亡
이선종류%붕체좌미%세포계,종류%세포증식%세포조망
Pancreatic neoplasms%Bortezomib%Cell line,tumor%Cell proliferation%Apoptosis
目的 观察硼替佐米对胰腺癌细胞BxPC3、SW1990增殖、凋亡的影响,探讨硼替佐米杀伤癌细胞的可能机制.方法 应用1、10、50、100、500 nmol/L及1、10 μmol/L的硼替佐米干预BxPC3、SW1990细胞,以硼替佐米未干预的细胞作为对照组.采用四甲基偶氮唑蓝(MTT)法检测细胞的增殖;流式细胞仪检测细胞凋亡;RT-PCR法检测Bak、Bax、Bcl-2、Bcl-xl、survivin mRNA表达;蛋白质印迹法检测pro-caspase-3、cleaved-caspase-3、Bax、Bcl-2、surviving蛋白表达.结果 大于50 nmol/L的硼替佐米干预胰腺癌BxPC3、SW1990细胞时,呈浓度及时间依赖性抑制细胞的增殖,其中硼替佐米对BxPC3细胞的生长抑制作用显著大于对SW1990细胞的作用,两者差异有统计学意义(P值均＜0.05).100 nmol/L硼替佐米干预组BxPC3和SW1990细胞凋亡率分别为(22.56±4.23)％和((12.71±2.23)％,显著高于对照组的(2.15±0.47)％和(2.32±0.54)％(P值均＜0.05),且BxPC3细胞的凋亡率显著高于SW1990细胞(P＜0.05).100 nmol/L硼替佐米干预48 h后BxPC3和SW1990细胞的Bak mRNA表达无显著变化,Bax mRNA及蛋白表达显著增加(P值均＜0.05),Bcl-2 mRNA和蛋白表达及Bcl-xl mRNA表达均减少(P值均＜0.05).survivin mRNA和蛋白表达在BxPC3细胞中均减少,而在SW1990细胞中均增加(P值均＜0.05).2株细胞的pro-caspase-3蛋白表达减少,而cleaved-caspase-3蛋白表达增加(P值均＜0.05).结论 硼替佐米可抑制胰腺癌细胞BxPC3、SW1990的增殖、诱导凋亡,对BxPC3细胞的作用高于对SW1990细胞,其机制可能与激活线粒体内源性凋亡途径及survivin参与的肿瘤耐药相关.
目的 觀察硼替佐米對胰腺癌細胞BxPC3、SW1990增殖、凋亡的影響,探討硼替佐米殺傷癌細胞的可能機製.方法 應用1、10、50、100、500 nmol/L及1、10 μmol/L的硼替佐米榦預BxPC3、SW1990細胞,以硼替佐米未榦預的細胞作為對照組.採用四甲基偶氮唑藍(MTT)法檢測細胞的增殖;流式細胞儀檢測細胞凋亡;RT-PCR法檢測Bak、Bax、Bcl-2、Bcl-xl、survivin mRNA錶達;蛋白質印跡法檢測pro-caspase-3、cleaved-caspase-3、Bax、Bcl-2、surviving蛋白錶達.結果 大于50 nmol/L的硼替佐米榦預胰腺癌BxPC3、SW1990細胞時,呈濃度及時間依賴性抑製細胞的增殖,其中硼替佐米對BxPC3細胞的生長抑製作用顯著大于對SW1990細胞的作用,兩者差異有統計學意義(P值均＜0.05).100 nmol/L硼替佐米榦預組BxPC3和SW1990細胞凋亡率分彆為(22.56±4.23)％和((12.71±2.23)％,顯著高于對照組的(2.15±0.47)％和(2.32±0.54)％(P值均＜0.05),且BxPC3細胞的凋亡率顯著高于SW1990細胞(P＜0.05).100 nmol/L硼替佐米榦預48 h後BxPC3和SW1990細胞的Bak mRNA錶達無顯著變化,Bax mRNA及蛋白錶達顯著增加(P值均＜0.05),Bcl-2 mRNA和蛋白錶達及Bcl-xl mRNA錶達均減少(P值均＜0.05).survivin mRNA和蛋白錶達在BxPC3細胞中均減少,而在SW1990細胞中均增加(P值均＜0.05).2株細胞的pro-caspase-3蛋白錶達減少,而cleaved-caspase-3蛋白錶達增加(P值均＜0.05).結論 硼替佐米可抑製胰腺癌細胞BxPC3、SW1990的增殖、誘導凋亡,對BxPC3細胞的作用高于對SW1990細胞,其機製可能與激活線粒體內源性凋亡途徑及survivin參與的腫瘤耐藥相關.
목적 관찰붕체좌미대이선암세포BxPC3、SW1990증식、조망적영향,탐토붕체좌미살상암세포적가능궤제.방법 응용1、10、50、100、500 nmol/L급1、10 μmol/L적붕체좌미간예BxPC3、SW1990세포,이붕체좌미미간예적세포작위대조조.채용사갑기우담서람(MTT)법검측세포적증식;류식세포의검측세포조망;RT-PCR법검측Bak、Bax、Bcl-2、Bcl-xl、survivin mRNA표체;단백질인적법검측pro-caspase-3、cleaved-caspase-3、Bax、Bcl-2、surviving단백표체.결과 대우50 nmol/L적붕체좌미간예이선암BxPC3、SW1990세포시,정농도급시간의뢰성억제세포적증식,기중붕체좌미대BxPC3세포적생장억제작용현저대우대SW1990세포적작용,량자차이유통계학의의(P치균＜0.05).100 nmol/L붕체좌미간예조BxPC3화SW1990세포조망솔분별위(22.56±4.23)％화((12.71±2.23)％,현저고우대조조적(2.15±0.47)％화(2.32±0.54)％(P치균＜0.05),차BxPC3세포적조망솔현저고우SW1990세포(P＜0.05).100 nmol/L붕체좌미간예48 h후BxPC3화SW1990세포적Bak mRNA표체무현저변화,Bax mRNA급단백표체현저증가(P치균＜0.05),Bcl-2 mRNA화단백표체급Bcl-xl mRNA표체균감소(P치균＜0.05).survivin mRNA화단백표체재BxPC3세포중균감소,이재SW1990세포중균증가(P치균＜0.05).2주세포적pro-caspase-3단백표체감소,이cleaved-caspase-3단백표체증가(P치균＜0.05).결론 붕체좌미가억제이선암세포BxPC3、SW1990적증식、유도조망,대BxPC3세포적작용고우대SW1990세포,기궤제가능여격활선립체내원성조망도경급survivin삼여적종류내약상관.
Objective To investigate the effect of bortezomib on proliferation and apoptosis of pancreatic cancer cell lines BxPC3,SW1990 and explore possible mechanisms of bortezomib's killing effect on cancer cells.Methods BxPC3,SW1990 cells were treated by using 1,10,50,100,500 nmol/L and 1,10 μmol/L of bortezomib,and cells without bortezomib treatment were considered as control group.The cell proliferation was determined by MTF assay,and apoptosis was determined by flow cytometry.Bak,Bax,Bcl2,Bcl-xl,survivin mRNA expressions were measured by RT-PCR,and Western blot was applied to determine the expressions of pro-caspase-3,cleaved-caspase-3,Bax,Bcl-2,surviving protein.Results When bortezomib concentration was higher than 50 nmol/L,it inhibited the proliferation of two cell lines in a dose and time-dependent manner.And with the same treatment the rate of proliferation inhibition of BxPC3 cells by bortezomib was greater than that of SW1990 cells,and the difference between the two cell lines was statistically significant (P ＜0.05).Apoptosis rates in the groups of BxPC-3 and SW1990 cells treated by 100 nmol/L bortezomib were (22.56 ± 4.23) ％ and (12.71 ± 2.23) ％,which were significantly higher than those in control group (2.15 ± 0.47) ％ and (2.32 ± 0.54) ％,P ＜ 0.05).In addition,apoptosis rate of BxPC3 cells was significantly higher than that of SW1990 cells (P＜0.05).Bak mRNA expression of BxPC3 and SW1990 cells after 100 nmol/L bortezomib treatment were not significantly changed,but the expression of Bax mRNA and protein was significantly increased (P ＜0.05).Bcl-2 mRNA and protein,as well as Bcl-xl mRNA expressions was significantly decreased (P ＜0.05).The expression of survivin mRNA and protein in BxPC3 cells were decreased,but were increased in SW1990 cells(P ＜0.05).The expression of pro-caspase-3 protein in the two cell lines was decreased,while the expression of cleaved-caspase-3 protein was increased (P ＜0.05).Conclusions Bortezomib can inhibit the proliferation of pancreatic cancer cell Iines BxPC-3 and SW1990 and induce apoptosis,and the effect on BxPC3 cells is more than that on SW1990 cells.The mechanism may depend on activation of the mitochondrial pathway of apoptosis,and be related to survivin-involved drug-resistance.
