二氢青蒿素对胰腺癌生长的抑制作用

目的 探讨二氢青蒿素在体外、体内对胰腺癌的生长抑制作用.方法 通过MTT法检测二氢青蒿素对胰腺癌细胞株生长的抑制作用,Annexin V-FITC/PI染色流式细胞术检测细胞凋亡;Western blot检测BxPC-3细胞中增殖、凋亡相关蛋白的表达.监测给药后胰腺癌裸鼠移植瘤体积的变化,并通过对肿瘤组织标本Ki-67染色和TUNEL染色检测肿瘤细胞增殖和凋亡情况.结果 MTT结果显示,二氢青蒿素可抑制体外培养的胰腺癌细胞BxPC-3和AsPC-1的增殖,诱导细胞凋亡,且呈剂量依赖性.二氢青蒿素亦可通过抑制增殖和诱导凋亡而抑制胰腺癌的体内生长.Western blot检测BxPC-3细胞中蛋白的表达水平,结果显示二氢青蒿素上调增殖相关蛋白p21WAF1、下调PCNA的表达;上调凋亡相关蛋白Bax、下调Bcl-2的表达,且可增加caspase-9的活化水平.结论 二氢青蒿素在体内外对胰腺癌均有抗肿瘤作用,是胰腺癌治疗的潜在药物.     

NF-κB P65亚基siRNA增强吉西他宾诱导胰腺癌细胞凋亡作用的实验研究

目的 探讨NF-κB P65亚基siRNA(NF-κB P65 siRNA)在体内外增强吉西他宾诱导胰腺癌细胞凋亡的作用及机制.方法 培养人胰腺癌细胞株BxPC-3和PANC-1,分为空白对照组、阴性干扰序列对照组、吉西他宾组、NF-κB P65 siRNA组和联合治疗组.MTT方法检测细胞增殖情况;Western blot方法检测NF-κB P65及凋亡相关蛋白的表达水平;Annexin V-FITC/PI染色流式细胞仪和激光共聚焦显微镜检测胰腺癌细胞凋亡情况;电泳凝胶迁移实验检测NF-κB的DNA结合活性.BxPC-3接种裸鼠皮下建立胰腺癌移植瘤模型.治疗后监测肿瘤体积;TUNEL染色检测移植瘤组织细胞凋亡指数.结果 转染后72 h,与其他组相比,联合治疗组显著降低了细胞活力指数(P<0.05),下调了Bel-2和proeaspase-3的表达水平,同时上调了Bax的表达水平;流式细胞仪检测结果显示,联合治疗组细胞凋亡率高于其他组(P<0.05);EMSA实验结果证实,NF-κB P65 siRNA组和联合治疗组NF-κB的DNA结合活性低于对照组(P<0.05).联合治疗能够通过诱导凋亡而抑制裸鼠移植瘤生长(P<0.01).结论 NF-κB P65 siRNA可以通过抑制NF-κB的DNA结合活性,调控凋亡相关蛋白的表达水平,激活线粒体凋亡途径,从而增强吉西他宾对胰腺癌细胞的促凋亡作用.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

二氢青蒿素对胰腺癌生长的抑制作用

Objective To investigate the anti-tumor activity of dihydroartemisinin in pancreatic cancer in vitro and in vivo. Methods For cultured cells,cell growth was determined by the MTT assay and apoptosis was evaluated by flow cytometry analysis stained with Annexin V-FITG/PI. The protein expression in BxPC-3 cells was analyzed by Western blot assay. BxPC-3 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors and the tumor volume was monitored after exposure to dihydroartemisinin. Ki-67 staining and TUNEL assay were used to assess tumor cell proliferation and apoptosis in tumor tissue. Results After treatment by dihydroartemisinin in vitro, the proliferative inhibition rates of pancreatic cancer cells BxPC-3 and AsPC-1 reached up to (76.2 ± 3.5) % and (79.5 ± 2.9) %, and the apoptosis rates were up to (55.5 ± 3.2)% and (40.0 ± 3.5)%, the differences were significantly (P ＜ 0.01) compared with control [(2.0 ± 1.3) % and (0.9 ± 0.4) %]. Dihydroartemisinin inhibited the growth of pancreatic xenograft tumors in nude mice. The proliferation index and apoptusis index were (49.1 ± 3.9)% and (50.2 ± 4.4)% respectively in dihydroarternisinin 50 mg/kg treatment group, compared to those of (72.1 ± 3.3) % and (9.4 ± 2.9) % in control, the differences were significantly (P ＜0.01). Western blot assay indicated that dihydroartemisinin up-regulates expression of proliferation-associated protein p21WAF1 and down-regulates expression of PCNA, increases expression of apoptosis-associated protein Bax and decreases expression of Bcl-2 and activates caspase-9 in BxPC-3 cells. Conclusions Dihydroartemisinin exerts anti-tumor activity in pancreatic cancer both in vitro and in vivo by proliferation inhibition and apoptusis induction. Dihydroartemisinin can be used as a potential anti-tumor drug in pancreatic cancer.     

胰腺癌患者循环microRNA及吉西他滨耐药的胰腺癌患者特异性microRNA表达谱分析

目的 检测胰腺癌患者血浆中表达差异的microRNA及对吉西他滨耐药的胰腺癌患者血浆中表达差异的microRNA,为寻找新的生物标志物用于胰腺癌的无创性诊断及吉西他滨的疗效预测提供新的思路.方法 采用qPCR方法检测并筛选出胰腺癌患者血浆中相比于健康人表达差异明显的microRNA,以及对吉西他滨耐药的胰腺癌患者血浆中相比于对吉西他滨不耐药的胰腺癌患者表达差异明显的microR-NA.结果 胰腺癌患者相较于健康人存在表达差异的microRNA有28个,对吉西他滨耐药的胰腺癌患者相较于不耐药患者存在表达差异的microRNA有28个.结论 胰腺癌患者血浆中microRNA表达与正常人群有较显著的差异,胰腺癌患者中对吉西他滨耐药和不耐药者血浆microRNA也存在较显著差异,其有可能作为胰腺癌的诊断及疗效预测提供潜在的生物标志物.     

吉西他滨诱导胰腺癌细胞ABCG2表达其化疗耐药的关系研究

目的：探讨吉西他滨诱导胰腺癌细胞ABCG2的表达及与其化疗耐药的关系。 方法：吉西他滨不同浓度、不同时间作用胰腺癌SW1990细胞后,用CCK-8法检测细胞的增殖抑制率,并计算吉西他滨不同作用时间的半数抑制浓度（IC50）;根据IC50值选择合适浓度的吉西他滨,作用SW1990细胞24、48、72 h后,用流式细胞仪检测细胞凋亡率,并用Western blot和RT-PCR法检测ABCG2蛋白与mRNA的表达。 结果：吉西他滨作用后,SW1990细胞增殖明显抑制,并呈浓度与时间依赖性,但IC50值呈时间依赖性增加（均P<0.05）;3.9 mg/mL吉西他滨作用24、48、72 h后,SW1990细胞总凋亡率逐渐增加,但晚期凋亡率呈降低趋势,ABCG2蛋白与mRNA的表达明显升高,并呈时间依赖性（均P<0.05）。 结论：吉西他滨能抑制胰腺癌细胞SW1990的增殖,但作用呈时间依赖性减弱,这可能与其诱导ABCG2上调表达有关。     

吉西他滨耐药人胰腺癌细胞株的建立及其与肿瘤干细胞的相关性研究

目的 建立吉西他滨耐药人胰腺癌细胞株SW1990/GZ,并探讨SW1990/GZ和胰腺癌肿瘤干细胞的相关性.方法 应用间歇浓度梯度倍增法建立吉西他滨耐药人胰腺癌细胞株SW1990/GZ;倒置显微镜下观察细胞形态;MTT法计算耐药指数(RI);荧光定量PCR检测ABCB1、ABCC1及ABCG2基因的表达水平;裸鼠皮下种植瘤试验观察SW1990和SW1990/GZ的成瘤能力;流式细胞仪通过侧群细胞(SP)法和表面特异抗原标记法(CIM4+CD24+)检测肿瘤干细胞含量.结果 在形态学上,SW1990/GZ较SW1990发生明显改变;SW1990/GZ的耐药指数是亲代SW1990的77.2倍;与亲代SW1990相比,耐药株SW1990/GZ中ABCB1、ABCC1及ABCG2的表达水平明显增高(P<0.01),裸鼠皮下成瘤能力增强(P<0.01);耐药株SW1990/GZ中SP细胞比例为(11.0±1.0)%,亲代SW1990中SP细胞比例为(4.6±0.9)%,CD44+CD24+细胞在两者中的比例分别为(8.7±0.8)%和(1.1±0.4)%(P<0.01).结论 吉西他滨耐药胰腺癌细胞株SW1990/GZ能高效富集胰腺癌肿瘤干细胞,CD44与胰腺癌获得性耐药关系密切,可能为克服胰腺癌获得性耐药提供新的治疗靶点.     

酸性神经磷脂酶及神经酰胺在吉西他滨诱导胰腺癌PANC-1细胞耐药中的作用

目的 建立对吉西他滨耐药的胰腺癌PANC-1/Gem细胞株,检测诱导耐药前后该细胞株生物学特性的变化.探讨吉西他滨诱导胰腺癌耐约的可能机制.方法 通过逐渐增加培养基中吉西他滨的浓度,建屯对吉西他滨耐药的胰腺癌PANC-1/Gem细胞株,TUNEL染色检测细胞株凋亡,MTT方法 检测胰腺癌PANC-1和胰腺癌PANC-1/Gem细胞的半数抑制浓度(IC_(50))和耐药指数(RI),Western印迹法检测酸性神经磷脂酶表达变化、二酰基甘油激酶(diacylglycerol kinase,DAGK)法检测神经酰胺含量变化.结果 经过24周成功诱导出对吉西他滨耐药的胰腺癌PANC-1/Gem细胞株,吉西他滨对PANC-1和PANC-1/Gem细胞的IC_(50)值分别为:亲本(8.13±0.85)μg/ml,24周(285.40±34.83)μg/ml,与亲本细胞相比耐药倍数为35.10倍,且凋亡率降低.Western印迹检测发现吉西他滨诱导24周的胰腺癌细胞PANC-1/Gem酸性神经磷脂酶的表达低于亲本细胞.两组细胞的神经酰胺含量分别为:亲本(364.95±46.11)pmol/mg protein,24周(120.61±20.07)pmol/mgprotein.结论 酸性神经磷脂酶的表达降低,导致神经酰胺含量减少可能是胰腺癌PANC-1细胞株对吉西他滨产生耐药的机制之一.     

乳香胶联合吉西他滨对人胰腺癌BxPc-3细胞的作用及机制初探

目的 研究乳香胶联合吉西他滨对体内、外培养的人胰腺癌BxPc-3细胞的作用及相关机制.方法 四唑氮蓝还原法检测BxPc-3细胞增殖,流式细胞仪检测细胞凋亡(PI单染色法).Western blot检测不同浓度的乳香胶联合吉西他滨(两药联合组)用药后BxPc-3细胞内测核转录因子-κB(NF-κB) p65亚基以及NF-κB抑制蛋白IkB、凋亡调节蛋白Bcl 2和Bax的表达水平的变化.建立人胰腺癌裸鼠皮下移植瘤模型,监测移植瘤体积变化.结果 两药联合组选择乳香胶(40 mg/L)和吉西他滨(10 mg/L)处理72 h后,抑制人胰腺癌细胞株BxPc-3细胞的作用显著优于单独使用(P＜0.01);凋亡比例(45.13±4.01)明显高于乳香胶组或吉西他滨组(P＜0.01)和对照组(5.07±1.37,P＜0.01);乳香胶或两药联合可抑制NF-κB的活性;对人胰腺癌BxPc-3细胞作用48 h后,Bcl-2的蛋白表达明显低于吉西他滨组和对照组(P＜0.01).而IkB和Bax蛋白的表达显著高于吉西他滨组和对照组(P＜0.01).与对照组比较,乳香胶或两药联合组可显著抑制裸鼠胰腺癌皮下移植瘤生长(P＜0.05).结论 乳香胶可通过抑制NF-κB的活性,增加IkB和Bax蛋白表达、抑制Bcl-2的蛋白表达,发挥抗肿瘤作用,并增强吉西他滨的药效.     

多药耐药相关基因及其蛋白在介导胰腺癌细胞原发性耐药中的作用及调控

目的 检测多药耐药相关基因（MDR1）及其蛋白（P-gP）在胰腺癌细胞株的表达,初步探讨胰腺癌发生先天性耐药的机理。方法 选择8株人胰腺癌细胞株,采用RT-PCR方法检测MDR1mRNA在胰腺癌细胞中的表达;通过免疫细胞化学方法检测P-gP的表达;以流式细胞仪检测胰腺癌细胞对罗丹明的外排情况,评价P-gP泵功能;最后通过P-gP抑制物维拉帕米与化疗药物联合应用,检测其对胰腺癌细胞的杀伤作用。结果 MDR1mRNA在胰腺癌细胞株SW1990中的表达最高,除PCT-2未检测到MDR1的表达外,其余6株细胞均有MDR1的微弱表达;免疫细胞化学染色结果证实P-gP在SW1990中的表达明显高于其他细胞株。57．9％±5．4％的SW1990细胞内有罗丹明蓄积,而在P-gP阴性对照细胞中,99．5％±3．3％的细胞内存在罗丹明的积聚,两者差异有统计学意义（P〈0．05）。P-gP抑制物维拉帕米与阿霉素或表阿霉素联合应用时可以部分逆转肿瘤细胞对化疗药物的耐药性。结论 MDR1及P-gP在人胰腺癌细胞中存在一定量的表达;维拉帕米联合阿霉素可以明显抑制P-gP阳性细胞的生长。     

Smac/DIABLO诱导胰腺癌细胞凋亡并增加对TRAIL及吉西他滨化疗的敏感性

目的 探讨外源性Smac/DIABLO对人胰腺癌SW1990细胞生物学特性和对TRAIL及吉西他滨化疗敏感性的影响. 方法 利用脂质体2000介导Smac/DIABLO基因转染胰腺癌SW1990细胞获得细胞SW1990/Smac,转染空载体为对照组(SW1990/neo);在不同浓度和时间下以肿瘤坏死因子相关凋亡诱导配体(TRAIL)和吉西他滨处理2组细胞株并分为TRAIL组、吉西他滨组和联合组.MTT法检测细胞株的生长抑制率,流式细胞仪检测细胞凋亡率及凋亡细胞形态,Western blot检测凋亡相关蛋白Smac/DIABLO、抑制凋亡蛋白XIAP、细胞色素C及细胞凋亡因子caspase-3的表达.结果 转染Smac/DIABLO的细胞生长明显落后于转染空载体的细胞.TRAIL浓度为200、500、1000、2500 ng/ml时,作用24 h对SW1990/neo和SW1990/Smac细胞的抑制率分别为11.11％、46.03％、67.08％、76.19％及22.11％、42.67％、56.63％、67.6％ (P ＜0.05).吉西他滨浓度分别为10、20、40、60μmol/L作用24 h对SW1990/neo和SW1990/Smac的抑制率分别为15.2％、34.6％、55.16％、76.4％和22.65％、36.85％、55.11％、79.99％ (P＜0.05).以TRAIL(500 ng/ml)、吉西他滨(20 μmol/L)及TRAIL(500 ng/ml)+吉西他滨(20 μmol/L)作用24 h后,SW1990/neo和SW1990/Smac细胞的凋亡率分别为5.64％、15.30％、27.27％和20.37％、23.27％、67.30％ (P ＜0.05).SW1990/Smac细胞在TRAIL及吉西他滨作用后,细胞内促凋亡蛋白Smac/DIABLO、细胞色素C及caspase-3活化片段表达均显著升高,而抑制凋亡蛋白XIAP表达显著降低(P＜0.05).结论 Smac/DIABLO可诱导SW1990细胞的凋亡、抑制细胞增殖,并增强胰腺癌细胞对TRAIL及吉西他滨的化疗敏感性,其机制可能与Smac/DIABLO、细胞色素C、XIAP及caspase-3的活性有关.     

高强度聚焦超声联合健择治疗晚期胰腺癌的临床研究

目的探讨高强度聚焦超声（HIFU）联合健择治疗晚期胰腺癌的有效性。方法将46例不能切除的晚期胰腺癌无黄疸病人随机分为：HIFU联合健择组24例，单用健择组22例。全部病例随访2～18个月。Kaplan-Meier法进行生存分析，计算6个月、12个月生存率和中位生存期；采用LogRank法进行组间比较。两组治疗后的疼痛缓解率采用X^2检验。结果HIFU联合健择组的24例病人中位生存时间为10．56个月，6个月和12个月的生存率分别是75．0％和30．9％；而单用健择组的22例病人中位生存时间为6．71个月，6个月和12个月的生存率分别是36．4％和13．0％。两组比较，HIFU联合健择组优于单用健择组，差异有显著性（P〈O．01）。HIFU联合健择组24例病人的疼痛缓解率（疼痛强度降低或消失，止痛剂减少或停止应用）87．5％；而单用健择组22例病人的缓解率为36．4％。两组比较，HIFU联合健择组明显优于单用健择组（P〈O．01）。结论HIFU联合健择组治疗晚期胰腺癌的疗效明显优于单用健择组，因此本方法有望成为治疗晚期胰腺癌的一种有效手段。     

人平衡核苷转运体1与胰腺癌化疗耐药性的关系

吉西他滨是临床胰腺癌化疗首选药物之一,然而患者对吉西他滨的化疗敏感性并不相同,部分患者对吉西他滨原发或继发耐药.人平衡核苷转运体1是介导吉西他滨进入肿瘤细胞的主要转运体,人平衡核苷转运体1在在不同患者肿瘤细胞中的表达差异性是造成胰腺癌吉西他滨化疗耐约的原因之一.     

胰腺癌细胞Panc-1中AKT2基因表达对吉西他滨敏感性的影响

目的 探讨胰腺癌细胞Panc-1中AKT2基因表达对吉西他滨敏感性的影响.方法 体外将AKT2特异性shRNA表达载体pAKT2-shRNA转染胰癌细胞株Panc-1.应用RT-PCR、Western blot检测转染shRNA后Panc-1细胞AKT2基因和蛋白的表达变化,应用MTT法检测干扰AKT2后Panc-1细胞对吉西他滨敏感性的变化.结果 转染pAKT2-shRNA后Panc-1细胞AKT2基因和蛋白的表达水平明显下降;吉西他滨对Panc-1细胞的半数抑制量从(1.96±0.22) mg/L降到(0.24±0.03)mg/L,Panc-1细胞对吉西他滨的敏感性明显增加.结论 用pAKT2-shRNA抑制AKT2基因表达,能增加人胰癌细胞株Panc-1对吉西他滨的敏感性.