[Purpose]To study the effect of hydroxyl camptothecinum (HCPT)on human pancreatic cancer cell proliferation and apoptosis (mainly early stage) in vitro.[Methods]SW1990 cell line were treated with HCPT at different concentrations.Cell proliferation were evaluated by MTT and apoptosis by Annexin-V-fluos stainging,electric microscopy,flow cytometry,bcl-2 immunocytochemical staining in vitro.[Results]① MTT cell proliferation detection:The rate of proliferation in each group of 3.125μ g/ml～ 100μ g/ml was significantly lower than that in the control group after being treated with HCPT 24 hours.② TUNEL and flow cytometry detection:The apoptosis rate of pancreatic cell was between 13.2％ ～ 15.3％ after being treated with HCPT at the concentration of 0.05mg/ml for 20 hours,and at 0.1mg/ml.The rate was between 26.1％ ～ 30.4％ .③ Annexin-V-fluos staining and electric microscopy detection:The expression of apoptosis was mainly at the early morphological stage at the concentration of 0.05mg/ml and the apoptosis rate of early stage about 15％ .④ The expression of bcl-2 immunocytochemical detection:The expression of bcl-2 was less than the control after being treated with HCPT 0.05mg/ml for 24 hours.[Conclusion]HCPT can inhibited the proliferation of SW1990 in vitro,partially by the induction of apoptosis,and HCPT is a potential agent for pancreatic cancer chemotherapy.

Objective: To study the effect of hydroxyl camptothecinum (HCPT) on proliferation and apoptosis of human pancreatic cancer cell in vitro . Methods: After treated with HCPT at different concentrations (1.562 5, 3.125, 6.25, 12.5 , 25, 50, 100 μg/ml) for 20 24 h, the cultured SW 1990 cell proliferation were evaluated by MTT, and apoptosis by Annexin Ⅴ fluos staining, electronic microscopy, flow cytometry, bcl 2 immunocytochemical staining and in situ TUNEL (TdT mediated dUTP X nick end labeling). Results: (1) The rate of proliferation in each group of 3.125 100 μg/ml was significantly lower than that in the control group ( P 0.01) after treated with HCPT for 24 hours. (2) The apoptosis rate of pancreatic cell was 13.2% and 15.3% after treated with HCPT at the concentration of 50 μg/ml for 20 hours, the rate was 26.1% and 28.4% at 100 μg/ml. (3)Annexin Ⅴ fluos staining and electronic microscopy detection showed the expression of apoptosis was mainly at the early morphological stage at the concentration of 50 μg/ml and the apoptosis rate of early stage was 18.4%. (4) The expression of bcl 2 were less than the control after treated with HCPT 50 μg/ml for 24 hours. Conclusion: HCPT can depress the proliferation of SW 1990 in vitro , partially through the induction of apoptosis, and HCPT is a potential agent for pancreatic cancer chemotherapy. [

目的 探讨基因重组转化生长因子α-绿脓杆菌外毒素融合蛋白(TGFα-PE40,简称TP40)对人低分化胃癌细胞生长的导向抑制作用.方法 用免疫组化LSAB法检测人低分化胃癌MKN-45细胞表皮生长因子受体(EGFR)的表达,用结晶柴染色法和3H-亮氨酸掺入法检测TP40对MNK-45细胞生长及蛋白质合成的抑制,用过量EGF竞争拮抗TP40的细胞毒作用.结果 MKN-45细胞免疫组化显示出大量棕黄色的EGFR阳性染色.TP40浓度为1～100ng/ml时,对MKN-45细胞长生及蛋白质合成呈剂量依赖性抑制.过量EGF能完全拮抗TP40的细胞毒作用.结论 人低分化胃癌MKN-45细胞能高水平地表达EGFR,重组TP40对MKN-45细胞生长具有较强的抑制作用,作用部位为细胞的EGFR.

目的:探讨基因重组转化生长因子α-绿脓杆菌外毒素融合蛋白(TGFα-PE40,简称TP40)对人肺腺癌细胞生长的导向抑制作用.方法:用免疫组化LSAB法检测人肺腺癌SPC-A-1细胞表皮生长因子受体(EGFR)的表达,用结晶紫染色法和3H-亮氨酸掺入法检测TP40对SPC-A-1细胞生长及蛋白质合成的抑制,用过量EGF竞争拮抗TP40的细胞毒作用.结果:SPC-A-1细胞免疫组化显示出大量棕黄色的EGFR阳性染色.TP40浓度为1～100ng/ml时,对SPC-A-1细胞生长及蛋白质合成呈剂量依赖性抑制.过量EGF能完全拮抗TP40的细胞毒作用.结论:人肺腺癌SPC-A-1细胞能高水平地表达EGFR,重组TP40对SPC-A-1细胞生长具有较强的抑制作用,作用部位为细胞的EGFR.

目的:观察动脉成形术对血管平滑肌细胞(SMC)表皮生长因子受体(EGFR)表达的影响,探讨EGFR与再狭窄的关系.方法:于兔右颈总动脉成形术前及术后24,48,72h,7d和14d,取实验动脉段用免疫组化LSAB法检测EGFR的表达.结果:正常颈总动脉中膜未见EGFR阳性表达;动脉成形术后24h,中膜部分SMC显示EGFR阳性表达;48和72h中膜EGFR阳性表达均进一步增加;7和14d动脉中膜EGFR阳性表达的SMC均较72h显著减少,但新生内膜中增殖的SMC均显示大量EGFR阳性表达.阳性染色以胞膜处最明显.结论:动脉成形术使血管SMC EGFR表达显著增加,再狭窄形成过程中SMC增殖与EGFR有关.

目的:探讨转化生长因子α(TGFα)-绿脓杆菌外毒素(PE40)融合蛋白(TP40)对血管平滑肌细胞(SMC)增殖的抑制作用.方法:用免疫组化ABC法检测原代培养增殖期及静止期SMC表皮生长因子受体(EGFR)的表达,用结晶紫染色法和3H-亮氨酸掺入法检测TP40对增殖期和静止期SMC增殖及蛋白质合成的抑制,用过量EGF竞争抑制TP40的细胞毒作用.结果:增殖期SMC能高水平表达EGFR,静止期SMC表达水平较低.TP40浓度为0.1和1.0 ng/ml时,对增殖期SMC增殖及蛋白质合成的抑制作用与静止期SMC无差异(P>0.05);10及100 ng/ml时,对增殖期SMC增殖及蛋白质合成的抑制作用较静止期SMC强(P<0.01);过量EGF能完全抑制TP40的细胞毒作用.结论:TP40对增殖期SMC的增殖具有较强的抑制作用,对静止期SMC则影响较小,作用部位为细胞的EGFR.

目的:观察球囊损伤对动脉平滑肌细胞(SMC)表皮生长因子受体(EGFR)表达的影响,探讨EGFR与再狭窄的关系.方法:于兔右颈总动脉球囊损伤前及损伤后24 h、48 h、72 h、7天和14天,取实验动脉段用免疫组化LSAB法检测EGFR的表达.结果:正常颈总动脉中膜未见EGFR阳性表达,动脉球囊损伤后24 h中膜部分SMC显示EGFR阳性表达,48 h中膜呈EGFR阳性表达的SMC较24 h增加,72 h又较48 h进一步增加;7天和14天动脉中膜EGFR阳性表达的SMC均较72 h显著减少,但新生内膜中增殖的SMC显示出大量的EGFR阳性表达.阳性染色主要定位于SMC的胞膜.结论:球囊损伤使动脉EGFR表达显著增加,EGFR与再狭窄形成过程中SMC增殖有关.