目的 探讨反应停对肺腺癌细胞株诱导的血管内皮细胞增殖的影响.方法 用噻唑兰法检测不同浓度反应停对血管内皮细胞株ECV304(ECV304细胞)、肺腺癌细胞株SPC-A1(SPC-A1细胞)及其诱导的血管内皮细胞ECV304增殖的影响.结果 反应停浓度为8.0和80.0 μg/ml时对ECV304细胞增殖有抑制作用(P＜0.05);不同浓度反应停对SPC-A1细胞增殖无影响;经8.0和80.0 μg/ml反应停作用后SPC-A1细胞条件培养液对ECV304细胞增殖有抑制作用(P均＜0.05),其细胞生长抑制率分别为5.19%、12.49%.结论 反应停对SPC-A1细胞诱导的ECV304细胞增殖有抑制作用.

目的:探讨BxPC-3条件培养液影响分化的PCI2细胞神经递质代谢的机制.方法:用去除IL-6的BxPC-3条件培养液处理分化的PC12细胞,采用流式细胞仪检测各组细胞凋亡率、HPLC检测上清和细胞裂解液中DA、5-HT和NE的含量.在不合血清的条件培养液中加入 IL-6(0.01 mg/L,0.1 mg/L,0.25mg/L,0.5 mg/L,1 mg/L,1.5 mg/L,2 mg/L),分别收集上清和细胞裂解液,采用HPLC检测DA和NE的含量.结果:各组细胞凋亡率之间差别无统计学意义,条件培养液中加入IL-6抗体后,上清中DA和NE的含量明显增加,而细胞内DA和NE代谢差异无统计学意义,在加入不同剂量IL-6后,随着浓度的增加,DA和NE的代谢近似呈现剂量依赖性降低,当IL-6浓度为1 mg/L时DA和NE的变化有统计学意义.结论:BxPC-3条件培养液主要通过IL-6影响分化的PC12神经递质代谢.

目的:研究晚期消化道癌症患者抑郁与免疫抑制酸性蛋白（immunosupressive acidic protein,IAP）等之间关系,并观察抗抑郁治疗后上述指标的变化,探讨抑郁与免疫功能之间关系.方法:采用Zung量表对晚期消化道癌症患者43例行抑郁测评的同时采用单向免疫琼脂扩散法检测外周血IAP含量,采用流式细胞仪检测外周血CD3,CD4,CD8和NK细胞百分比,根据Zung测评结果,以百忧解对抑郁患者进行干预,抑郁纠正的同时再次检测上述指标结果:本组患者抑郁的发生率为44%;抑郁评分与IAP呈正相关（r=0.549,P＜0.01）,与CD3,CD4呈负相关（r=-0.163,P＜0.001;r= -0.163,P＜0.001）,与NK,CD8无相关（r=0.262,P＞0.05:r=0.611,P＞0.05）;抑郁组与非抑郁组数据分析表明IAP,CD3和CD4有显著差异（IAP:908.33±275.75 mg/L vs 537.60±282.64 mg/L,P＜0.01;CD3:67.05±13.12 vs 73.27±8.03.P＜0.05:CD4:36.68±11.29 vs 43.72±9.16.P＜0.05）;抗抑郁治疗后IAP变化明显（P＜0.05）,CD3,CD4,CD8,NK无明显变化（均P＞0.05）.结论:老年晚期消化道癌症患者抑郁与免疫功能相关,伴发抑郁的患者免疫功能低下,抗抑郁治疗后免疫功能改善.

BACKGROUND & OBJECTIVE:The malignant tumor tissues usually suffer from hypoxia due to its fast growth, which increases its invasive ability. This study was to explore changes of matrix metalloprotei-nases (MMPs) in tumor cells under hypoxia in vitro.METHODS:Zymography was used to determine the changes of activities of MMP-2 and MMP-9 in pancreatic cancer cell line SW1990 cultured under hypoxia. Western blot, and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect protein and mRNA levels of MMP-2 and MMP-9.RESULTS:Under normal circumstances, activities of MMPs in SW1990 cells were relatively low. When SW1990 cells were cultured under hypoxia, the activity of MMP-9 increased by about 5.5 times that in normal circumstances, while the activity of MMP-2 didn't change. The same changes in protein level of MMP-9 had been confirmed by Western blot, while protein level of MMP-2 didn't change. mRNA level of MMP-9 was higher in cells cultured under hypoxia for 12 h than in control cells and CoCl(2)-cultured tumor cells by 46.1% and 56.4%, but no change of MMP-9 was seen when hypoxia inducible factor-1alpha (HIF-1alpha) was produced in SW1990 cells stimulated by CoCl(2).CONCLUSIONS:Production of MMP-9 in SW1990 cells is enhanced significantly under hypoxia, and its activity altered correspondingly; but those of MMP-2 had no change. This alteration has no relation with enhancement of HIF-1alpha under hypoxia.

Objective To investigete the mechanism of thalidomide regulating angiogenesis in Lewis lung cancer model. Methods Mice model of Lewis lung cancer was established, treated with different doses of thalidomide。Normal saline and TNP470 were as negative and positive factor。 Masses of tumor were weighed。 bFGF and MMP-2 , MVD were evaluated by CD34 antibody with immunohistochemistry, the level of bFGF and MMP-2 was examined by Western Blot method. Results Thalidomide(200 or 400 mg/kg)inhibitd the growth of Lewis lung cancer, down regulated the expression of bFGF and MMP-2, and reduced MVD in Lewis lung cancer model. Conclusion Thalidomide can inhibit the angiogenesis inhibiting vascular growth factors bFGF and MMP-2 in Lewis lung cancer.

目的探讨羟基喜树碱对人肝癌细胞株SMMC-7721增殖的影响及其诱导凋亡的作用.方法以不同浓度的羟基喜树碱在体外作用于人肝癌细胞株SMMC-7721,采用MTT法检测细胞增殖;化学荧光法检测Bcl-2的细胞表达;电镜观察、流式细胞仪与原位末端标记方法检测细胞凋亡情况.结果与空白对照组比较,羟基喜树碱浓度在3.125μg/mL～100μg/mL组,对肿瘤细胞的增殖抑制率显著增高(P＜0.01);在0.05mg/mL组出现细胞早期凋亡现象,Bcl-2表达明显减少;在0.1mg/mL组形成凋亡小体;细胞凋亡比例随羟基喜树碱浓度增加而增高.结论羟基喜树碱在体外可抑制人肝癌细胞株SMMC-7721增殖,其作用机制可能系通过抑制Bcl-2表达诱导细胞凋亡.

Objective:To investigate the inhibitory effect of panaxoside Rg3 on vascular endothelial cell (VEC) proliferation induced by lung adenocarcinoma.Methods:Inhibitory effects of panaxoside Rg3 on proliferation of VEC,lung adenocarcinoma SPC A 1 cell and VEC proliferation induced by SPC A 1 cell were analyzed with MTT method.Results:Panaxoside Rg3 in the range of 0.0313～0.5 mmol·L -1 had no direct influence on VEC and SPC A 1 cell proliferation ( P 0.05),and had also no influence on VEC proliferation induced by SPC A 1 cell conditional cultured medium acted with 0.0313～ 0.5 mmol·L -1 of panaxoside Rg3 ( P 0.05),but panaxoside Rg3 in the range of 0.125～ 0.5 mmol·L -1 inhibited VEC proliferation induced by conditional cultured medium of SPC A 1 cell ( P 0.01),the inhibitory rate was 16.44%～57.53%.Conclusion:Panaxoside Rg3 can inhibit VEC proliferation induced by conditional cultured medium of lung adenocarcinoma cell.

新生血管形成是实体瘤快速增长所必须的先决条件之一,一方面,新生血管可以为肿瘤细胞带来丰富的氧气和养料,并将代谢产物带走;另一方面,肿瘤细胞通过新生的血管向远处转移,因此探讨肿瘤新生血管形成机制已成为目前肿瘤研究领域的又一个重要课题.

Objective: To study the mechanism of inhibitory ef fect of ginsenoside Rg3 on tumor growth. Methods: The chick chor ioallantoic membrane(CAM) test and Lewis lung carcinoma model were used to inves tigate the inhibitory effect of Rg3 on tumor angiogenesis. Results: Rg3(0.1 or 0.5 mmol/L) obviously inhibited angiogenesis in the CAM. Treatmen t with Rg3 in vivo obviously inhibited Lewis lung carcinoma growth with the inhibition ratio from 23% to about 47%. We also observed that the angiogenesis in implanted Lewis lung carcinoma tissue decreased obviously after treated wit h Rg3 (5, 10, 20 mg/kg). Conclusion: Rg3 can obviously inhibit t he growth of Lewis lung cancer, the inhibitory effect partially due to the effec t of Rg3 inhibiting neovascularization induced by malignant tumor.

Objective: To design the ribozymes to cleave human TIMP 1 mRNA, and embed them into U 6snRNA to make them stable. Methods: Ribozymes were designed according to the “hammerhead structure” described by Symons.Computer was used to analyze the possible cleavage sites. Results: Three ribozymes targeting the nt123, nt299 and nt353 on TIMP 1 mRNA were designed. Embedding ribozyme in U 6snRNA had little effect on its binding with the substrate. Conclusion: Computer assisted design is indispensable in studying ribozyme. Embedding ribozymes in U 6snRNA may be a good way to solve the problems existing in ribozyme study. [

Objective To design the ribozymes which may cleave the mRNA of human TIMP-1. Establishing plasmids transcripting the ribozymes to produce abundant ribozymes. Methods The TIMP-1 mRNA was taken as the target RNA. Ribozymes were designed according to the hammerhead structure described by Symons. To establish clones of TIMP-1 ribozymes use P 1.5. Then obtain abundant ribozymes by transcripting. Results Three ribozymes targeting the nt 123, nt 299 and nt 353 on TIMP-1 mRNA were designed. The ribozyme gene has been synthesized, plasmids cloned and abundant ribozymes obtained. Conclusion Computer assisted design is indispensable in studying ribozyme. The establishment of plasmids transcripting ribozymes is the key step of studying their activity.

Objective: To study the effect of arsenic trioxide (As2O3) on human pancreatic cancer cell proliferation and apoptosis (mainly early stage) in vitro. Methods: SW1990 cells line were trea ted with As2O3 at different concentration. Cell proliferation was evaluated by MTT and apoptosis by Annexin-Ⅴ-fluostaining, electron-microscopy, flow cytometry and immunocytochemical staining of Bcl-2 and Bax. Results: As2O3 and cisplatin had the same cytotoxity on SW1990. The cytotoxic effe ct on tumor cell was produced by induction of apoptosis. Twelve hours after cult ure with 10 μg/ml As2O3, much more SW1990 cells went into apoptosis than t he control. The apoptosis rate reached 24% after 48 h with the similar concentra tion of As2O3. Immunohistochemical study revealed that the expression of Bcl -2 was decreased after treated with As2O3. Conclusion: As 2O3 can depress the proliferation of SW1990 in vitro, mainly through the i nduction of apoptosis, and it is a potential agent for pancreatic cancer chemoth erapy.

[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 研究羟基喜树碱体外诱导人肺癌细胞株SPC-A-1 凋亡的作用。方法 以不同浓度的羟基喜树碱处理在体外培养的肺癌细胞,以电子显微镜、流式细胞仪和原位末端标记分析来检测细胞凋亡。结果 在0.05 m g/m l浓度下,羟基喜树碱作用于体外培养的肺癌细胞20 h 后,肿瘤细胞的凋亡率在8.7 % ～10.4 % 之间,主要以早、中期的表现为主;在0.1 m g/m l浓度下,作用20 h 后,凋亡率为18.6% ,凋亡形态学表现主要以晚期为主。结论 羟基喜树碱可作为细胞凋亡诱导剂用于肺癌治疗