中国癌症杂志
中國癌癥雜誌
중국암증잡지
CHINA ONCOLOGY
2012年
11期
801-807
,共7页
二甲双胍%人胰腺癌细胞株Bxpc-3%增殖%凋亡
二甲雙胍%人胰腺癌細胞株Bxpc-3%增殖%凋亡
이갑쌍고%인이선암세포주Bxpc-3%증식%조망
Metformin%Human pancreatic cell line Bxpc-3%Proliferation%Apoptosis
背景与目的:近年研究发现二甲双胍对多种肿瘤具有抑制作用,部分临床研究也发现二甲双胍可以降低糖尿病并发胰腺癌的风险和降低胰腺癌的死亡危险.本研究从细胞水平着手研究二甲双胍对胰腺癌细胞增殖和凋亡的影响,并初步探讨其可能机制.方法:体外培养人胰腺癌细胞株Bxpc-3,予二甲双胍进行干预作为二甲双胍干预组,无药物组作为对照组.以MTT检测二甲双胍对Bxpc-3细胞存活率的影响,流式细胞术检测二甲双胍对Bxpc-3细胞周期的影响,流式细胞术及Hoechst?33258荧光染色法检测细胞凋亡,RT-PCR检测AMPKα1、Bax、Bcl-2、Caspase-3、Cyclin? D1? mRNA的表达.蛋白质印迹法(Western? blot)检测Cyclin? D1、Bax、Bcl-2、Caspase-3蛋白的表达.结果:与对照组相比,MTT检测结果显示,二甲双胍可以抑制人胰腺癌细胞株Bxpc-3的增殖,并呈时间-浓度依赖性(F=8.99、124和114.61,P<0.01);流式细胞术检测结果显示,二甲双胍干预组与对照组相比,G0/G1期细胞所占百分比增加(t=-8.71,P<0.01),S期(t=7.54,P<0.01)及G2/M期(t=7.00,P<0.01)细胞所占百分比减少;流式细胞术(早期凋亡率t=-2.68,晚期凋亡率t=1.29,总凋亡率t=-0.85)及Hoechst?33258荧光染色法(t=-0.46)结果显示,两组细胞凋亡率差异无统计学意义(P>0.05);RT-PCR结果显示,二甲双胍干预组Cyclin? D1? mRNA表达明显降低(t=4.96,P<0.01),AMPK?α1、Bax、Bcl-2、Caspase-3? mRNA表达与对照组相比,差异无统计学意义(t=1.68、-0.56、-1.80、0.67,P>0.05);Western?blot结果显示,二甲双胍干预组Cyclin?D1蛋白表达明显降低(t=7.02,P<0.01),Bax、Bcl-2、Caspase-3?蛋白表达与对照组相比,差异无统计学意义(t=-0.11、-0.20,?0.11,? P>0.05).结论:二甲双胍能显著抑制人胰腺癌细胞株Bxpc-3的增殖,机制主要与其阻滞细胞周期、下调Cyclin? D1表达有关;二甲双胍对Bxpc-3细胞的凋亡无明显诱导作用.
揹景與目的:近年研究髮現二甲雙胍對多種腫瘤具有抑製作用,部分臨床研究也髮現二甲雙胍可以降低糖尿病併髮胰腺癌的風險和降低胰腺癌的死亡危險.本研究從細胞水平著手研究二甲雙胍對胰腺癌細胞增殖和凋亡的影響,併初步探討其可能機製.方法:體外培養人胰腺癌細胞株Bxpc-3,予二甲雙胍進行榦預作為二甲雙胍榦預組,無藥物組作為對照組.以MTT檢測二甲雙胍對Bxpc-3細胞存活率的影響,流式細胞術檢測二甲雙胍對Bxpc-3細胞週期的影響,流式細胞術及Hoechst?33258熒光染色法檢測細胞凋亡,RT-PCR檢測AMPKα1、Bax、Bcl-2、Caspase-3、Cyclin? D1? mRNA的錶達.蛋白質印跡法(Western? blot)檢測Cyclin? D1、Bax、Bcl-2、Caspase-3蛋白的錶達.結果:與對照組相比,MTT檢測結果顯示,二甲雙胍可以抑製人胰腺癌細胞株Bxpc-3的增殖,併呈時間-濃度依賴性(F=8.99、124和114.61,P<0.01);流式細胞術檢測結果顯示,二甲雙胍榦預組與對照組相比,G0/G1期細胞所佔百分比增加(t=-8.71,P<0.01),S期(t=7.54,P<0.01)及G2/M期(t=7.00,P<0.01)細胞所佔百分比減少;流式細胞術(早期凋亡率t=-2.68,晚期凋亡率t=1.29,總凋亡率t=-0.85)及Hoechst?33258熒光染色法(t=-0.46)結果顯示,兩組細胞凋亡率差異無統計學意義(P>0.05);RT-PCR結果顯示,二甲雙胍榦預組Cyclin? D1? mRNA錶達明顯降低(t=4.96,P<0.01),AMPK?α1、Bax、Bcl-2、Caspase-3? mRNA錶達與對照組相比,差異無統計學意義(t=1.68、-0.56、-1.80、0.67,P>0.05);Western?blot結果顯示,二甲雙胍榦預組Cyclin?D1蛋白錶達明顯降低(t=7.02,P<0.01),Bax、Bcl-2、Caspase-3?蛋白錶達與對照組相比,差異無統計學意義(t=-0.11、-0.20,?0.11,? P>0.05).結論:二甲雙胍能顯著抑製人胰腺癌細胞株Bxpc-3的增殖,機製主要與其阻滯細胞週期、下調Cyclin? D1錶達有關;二甲雙胍對Bxpc-3細胞的凋亡無明顯誘導作用.
배경여목적:근년연구발현이갑쌍고대다충종류구유억제작용,부분림상연구야발현이갑쌍고가이강저당뇨병병발이선암적풍험화강저이선암적사망위험.본연구종세포수평착수연구이갑쌍고대이선암세포증식화조망적영향,병초보탐토기가능궤제.방법:체외배양인이선암세포주Bxpc-3,여이갑쌍고진행간예작위이갑쌍고간예조,무약물조작위대조조.이MTT검측이갑쌍고대Bxpc-3세포존활솔적영향,류식세포술검측이갑쌍고대Bxpc-3세포주기적영향,류식세포술급Hoechst?33258형광염색법검측세포조망,RT-PCR검측AMPKα1、Bax、Bcl-2、Caspase-3、Cyclin? D1? mRNA적표체.단백질인적법(Western? blot)검측Cyclin? D1、Bax、Bcl-2、Caspase-3단백적표체.결과:여대조조상비,MTT검측결과현시,이갑쌍고가이억제인이선암세포주Bxpc-3적증식,병정시간-농도의뢰성(F=8.99、124화114.61,P<0.01);류식세포술검측결과현시,이갑쌍고간예조여대조조상비,G0/G1기세포소점백분비증가(t=-8.71,P<0.01),S기(t=7.54,P<0.01)급G2/M기(t=7.00,P<0.01)세포소점백분비감소;류식세포술(조기조망솔t=-2.68,만기조망솔t=1.29,총조망솔t=-0.85)급Hoechst?33258형광염색법(t=-0.46)결과현시,량조세포조망솔차이무통계학의의(P>0.05);RT-PCR결과현시,이갑쌍고간예조Cyclin? D1? mRNA표체명현강저(t=4.96,P<0.01),AMPK?α1、Bax、Bcl-2、Caspase-3? mRNA표체여대조조상비,차이무통계학의의(t=1.68、-0.56、-1.80、0.67,P>0.05);Western?blot결과현시,이갑쌍고간예조Cyclin?D1단백표체명현강저(t=7.02,P<0.01),Bax、Bcl-2、Caspase-3?단백표체여대조조상비,차이무통계학의의(t=-0.11、-0.20,?0.11,? P>0.05).결론:이갑쌍고능현저억제인이선암세포주Bxpc-3적증식,궤제주요여기조체세포주기、하조Cyclin? D1표체유관;이갑쌍고대Bxpc-3세포적조망무명현유도작용.
? Background and purpose:In recent years, studies found that metformin could inhibit various kinds of tumor. And a few clinical studies also found that metformin could reduce the risk of diabetes combined with pancreatic cancer and could reduce the death risk of pancreatic cancer. So we proceed from a cellular level to investigate the effects of metformin on proliferation and apoptosis in human pancreatic cancer cell line Bxpc-3, and to appraise the possible potential mechanism.? Methods:Human pancreatic cancer cell line Bxpc-3 were cultured in vitro, and were treated with metformin as trial group or without metformin as control group. Survival rate of Bxpc-3 cells was determined by Methyl thiazolyl tetrazolium (MTT) assay, and the cell cycle changes were analyzed by flow cytometry (FCM). Apoptosis was determined by FCM and Hoechst 33258 fluorescence staining. Expressions of AMPK α1, Bax, Bcl-2, Caspase-3, Cyclin D1 mRNA were determined by RT-PCR. Protein expression of Cyclin D1, Bax, Bcl-2, Caspase-3 were determined by Western blot. Results:Compared with the control, metformin decreased the proliferation of Bxpc-3 cells in a dose- and time-dependent manner (F=8.99, 124, 114.61, P<0.01). Compared with the control, the proportion of the cells at G0/G1 stage in the metformin-treated was increased (t=-8.71, P<0.01), and the proportion of the cells at S (t=7.54, P<0.01) and G2/M (t=7.00, P<0.01) were decreased. The apoptosis rate of control group and metformin-treated group were not difference significantly (early apoptosis rate t=-2.68, late apoptosis rate t=1.29, total apoptosis rate t=-0.85). Expression of Cyclin D1 mRNA was down-regulated (t=4.96, P<0.01 ). Expression of AMPK αl, Bax, Bcl-2 and Caspase-3 mRNA were not altered significantly (t=1.68, -0.56, -1.80, 0.67, P>0.05). Protein expression of Cyclin D1 was down-regulated (t=7.024, P<0.01). Protein expression of Bax, Bcl-2 and Caspase-3 were not altered significantly (t=-0.11, -0.20, 0.11, P>0.05). Conclusion:Metformin can inhibit the proliferation of human pancreatic cancer cell line Bxpc-3 mainly by blocking the cell cycle at G0/G1 and down-regulating the expression of Cyclin D1. Metformin could not induce significantly apoptosis of human pancreatic cancer cell line Bxpc-3.
