基于活体成像的裸小鼠肺癌移植瘤模型建立及意义

目的建立可供生物学功能研究或药物评价的稳定表达绿色荧光蛋白的小鼠肺癌移植瘤模型。方法用脂质体将绿色荧光蛋白表达载体pEGFP-N1转染至人非小细胞肺癌细胞株NCI-H358,经G418筛选获得稳定表达绿色荧光蛋白的NCI-H358-GFP细胞,接种至裸小鼠皮下建立移植瘤模型。采用活体成像系统连续5周监测小鼠皮下NCI-H358-GFP细胞的动态生长情况。结果获得了在体外及裸小鼠体内均能够长期稳定表达绿色荧光蛋白的NCI-H358-GFP细胞。其接种后裸小鼠皮下全部成瘤,活体成像系统观察发现,随着NCI-H358-GFP细胞接种时间延长,移植瘤体积逐渐增大,荧光光子数逐渐增加。结论成功构建可用于活体成像的裸小鼠肺癌移植瘤模型;该模型可作研究肺癌发生发展机制及抗肿瘤药物的重要工具。     

南蛇藤乙酸乙酯提取物对荧光蛋白标记的HepG2细胞裸鼠移植瘤生长的抑制作用

目的 建立裸鼠原位人荧光蛋白肝癌移植瘤模型,采用活体荧光成像技术研究南蛇藤乙酸乙酯提取物对人肝癌生长的抑制作用.方法 BALB/c裸鼠肝脏接种荧光蛋白标记的人肝癌细胞(RFP-HepG2),建立肝癌动物模型并随机分为空白对照组(G1组)、奥沙利铂阳性对照组(G2组,25mg/kg)、南蛇藤小剂量治疗组(G3组,20mg/kg)、南蛇藤大剂量治疗组(G4组,40mg/kg)和南蛇藤预防治疗组(G5组,20mg/kg).治疗组自RFP-HepG2细胞接种第20天开始用药,每日给药1次,连续4周.奥沙利铂尾静脉注射给药,南蛇藤灌胃给药,G1组用等渗盐水替代.G5组自RFP-HepG2细胞接种第2天开始给药至治疗结束.活体荧光影像技术追踪肿瘤生长情况,测量移植瘤体积;治疗结束后切除肿瘤并称重.组间肿瘤体积和质量比较用t检验.结果 RFP-HepG2细胞接种第31天的影像扫描结果提示各组移植瘤体积出现差异,第45天达高峰,G1、G2、G3、G4、G5组的移植瘤体积分别为(803.1±512.3)mm3、(83.8±23.5)mm3、(852.7±502.6)mm3、(410.0±231.6)mm3、(120.5±60.1)mm3;G5组肿瘤体积较G1组明显减小(t=3.723,P＜0.01),与G2组间的差异无统计学意义(t=0.163,P＞0.05);G4组与G1组间的差异无统计学意义(t=2.156,P＞0.05),且均明显大于G2组(t=4.526,P＜0.05).G1、G2、G3、G4、G5组瘤体质量分别为(0.95±0.49)g、(0.36±0.09)g、(0.67±0.29)g、(0.48±0.15)g、(0.38±0.11)g;G2、G4和G5组瘤体质量明显小于G1组(t值分别为3.371、2.774和2.901,P值均＜0.05),G4组和G5组与G2组间差异无统计学意义(t值分别为1.735、0.259,P＞0.05),而G3组瘤体质量则明显高于G2组(t=2.684,P＜0.05).结论 大剂量南蛇藤对移植瘤具有明显的抑制作用,疗效稍低于奥沙利铂;预防用药组肿瘤的生长速度明显减弱,其作用与奥沙利铂相当.     

人参皂苷Rg3对小鼠肝癌血管形成的影响

目的：探讨人参皂苷Rg3抑制人肝癌组织血管生成的机制。方法：①选用昆明种小鼠6只（雌雄各半,供传代用）,昆明种小鼠140只（雌雄各半）按体重随机分为正常对照组（20只）,肝癌模型组（30只）,人参皂苷R船组（30只）,5-FU（5-氟尿嘧啶）组（30只）,人参皂苷Rg3联合5-FU组（30只）。采用小鼠肝癌细胞株H22肝内注射建立小鼠移植性肝癌模型的方法,将小鼠肝癌细胞株H22肝内注射24h后,人参皂苷Rg3组小鼠灌胃给药,5-FU组腹腔内注射给药,人参皂苷Rg3联合5-FU组小鼠灌胃和腹腔内注射给药,连续10d后处死各组全部小鼠并留取标本进行检测。②免疫组织化学染色观察各组小鼠肝组织血管内皮生长因子（VEGF）的表达。结果：5-FU组、人参皂苷Rg2组、人参皂苷Rg3联合5-FU组小鼠肝癌组织VEGF表达水平均低于肝癌模型组（均P〈0．05）;人参皂苷Rg3组和人参皂苷Rg3联合5-FU纽小鼠肝癌组织VEGF表达水平均低于5-FU组（均P〈0．05）;人参皂苷Rg3联合5-FU组小鼠肝癌组织VEGF表达水平与人参皂苷Rg3组比较,差异无显著性意义（P〉0．05）。结论：人参皂苷Rg3可抑制肝癌细胞血管生成,其可能机制是抑制VEGF表达。     

人参皂苷Rg3对人肝癌细胞Pim-3及Bad凋亡蛋白表达的影响

目的:研究人参皂苷Rg3对人肝癌细胞SMMC-7721中Pim-3及磷酸化Bad蛋白pBad(Ser112),pBad(Ser136)表达的影响.方法:用浓度为0、5、10、20、40和80 μmol/L的人参皂苷Rg3处理SMMC-7721细胞24 h后.采用四甲基偶氮唑盐(MTT)方法检测人参皂苷Rg3对SMMC-7721细胞增殖的抑制作用,倒置显微镜和流式细胞术观察人参皂苷Rg3对SMMC-7721细胞凋亡的诱导作用,Western blot方法检测不同浓度人参皂苷Rg3处理后SMMC-7721细胞中Pim-3,pBad(Ser112)和pBad(Ser136)的表达情况.结果:5、10、20、40和80μmol/L的人参皂苷Rg3对SMMC-7721细胞增殖的抑制率分别为4.69%、15.53%、22.17%、50.97%、61.65%.5.80 μmol/L的人参皂苷Rg3处理细胞呈现明显的凋亡形态学改变,80μmol/L人参皂苷Rg3处理SMMC-7721细胞24 h后,处理组比正常对照组凋亡明显增加,差异有统计意义(16.5%±4.3% VS 0.4%±1.3%,P<0.01).人参皂苷Rg3对细胞中Bad总蛋白的表达没有明显影响.Pim-3的表达随人参皂苷Rg3浓度的增加而逐渐减弱,而pBad(Ser112)的表达随人参皂苷Rg3浓度的增加而逐渐增强;pBad(Ser136)不表达,结论:人参皂苷Rg3的抗癌活性与其促进磷酸化Bad蛋白表达有关.     

人参皂苷Rg3联合索拉非尼对人肝癌细胞株侵袭与转移的影响

目的探讨人参皂苷Rg3联合索拉非尼对人肝癌细胞株侵袭与转移的影响。方法分别将人参皂甙Rg3和索拉非尼两种单药、两药联合方式作用于人肝癌细胞,观察其对细胞侵袭和转移的影响。结果作用1 h,索拉非尼组黏附抑制率显著高于人参皂苷Rg3组(P0.05或0.01),联合组显著高于索拉非尼组和人参皂苷Rg3组(P0.01);作用2 h,人参皂苷Rg3组抑制率显著高于索拉非尼组(P0.01),并且高于人参皂苷Rg3组作用1 h(P0.01);两药联合对肝癌细胞的黏附抑制率具有协同作用(Q=2.631.15)。索拉非尼单药及索拉非尼联合人参皂苷Rg3对肝癌细胞的迁移抑制率均显著高于人参皂苷Rg3单药作用(P0.01),并且两药联合的抑制率显著高于索拉非尼单药组(P0.01);两药联合对肝癌细胞的迁移抑制作用具有协同作用(Q=1.461.15)。索拉非尼组和联合组侵袭抑制率均显著高于人参皂苷Rg3组(P0.01),两药联合对肝癌细胞的侵袭抑制作用具有拮抗作用(Q=0.710.85)。与阴性对照组比较,人参皂苷Rg3组CD44V6、基质金属蛋白酶(MMP)-9表达水平显著下降,索拉非尼组和联合组血管内皮生长因子(VEGF)-C、CD44V6及MMP-9表达均显著下降(P0.05);与人参皂苷Rg3组比较,联合组VEGF-C及CD44V6水平显著下降;与索拉非尼组比较,联合组VEGF-C水平显著下降(P0.05)。结论人参皂苷Rg3、索拉非尼单药及两药联合对肝癌细胞SMMC-7721细胞株的黏附、迁移、侵袭具有一定的抑制作用,对细胞的黏附和迁移抑制作用两者均具有协同作用。     

人参皂苷Rg3联合索拉非尼对裸鼠肝癌移植瘤生长和血管生成的调控作用

目的:研究人参皂苷Rg3联合索拉非尼对裸鼠肝癌移植瘤生长及血管生成的影响及其机制.方法:构建裸鼠人肝癌移植瘤模型LCI-D20,将造模的26只裸鼠随机分成人参皂苷组(R组):人参皂苷Rg3 5 mg/kg,腹腔注射,1次/d;索拉非尼组(S组):索拉非尼30 mg/kg,灌胃,1次/d;联合组:人参皂苷Rg3 5 mg/kg+索拉非尼30 mg/kg;对照组:生理盐水腹腔注射,1次/d.治疗2 wk剥离瘤体称质量,计算抑瘤率;免疫组织化学法检测移植瘤组织微血管密度(microvascular density,MVD);ELISA法、Western blot法检测血管内皮生长因子(vascular endothelial growth factor,VEGF)、低氧诱导因子-1?(hypoxia inducible factor-1?,HIF-1?)和VEGFR-2的表达.结果:人参皂苷Rg3组的抑瘤率为20.6%,索拉非尼组的抑瘤率为34.74%,联合组的抑瘤率为48.64%,根据Weeb系数计算方法,人参皂苷Rg3与索拉非尼联合用药后肿瘤的生长率为51.36%,低于预期值51.81%,两者表现为协同作用.免疫组织化学法测定显示,3组的MVD均较对照组明显降低(P0.01),但3组间差异不明显(P0.05).ELLISA法结果显示:R组、S组及联合组血清VEGF水平均低于对照组(P0.01、P0.05及P0.01),联合组较S组水平更低(P0.05);R组及联合组血清HIF-1?水平明显低于对照组(P0.01),S组仅有降低的趋势(P0.05);联合组与S组间有明显差异(P0.05);3组血清VEGFR-2水平与对照组比较差异不明显(P0.05).Western blot法结果显示:3组VEGF、HIF-1?和VEGFR-2蛋白的表达均低于生理盐水组(P0.05),但联合组与R、S组之间均无明显差异(P0.05).结论:人参皂苷Rg3联合索拉非尼对裸鼠肝移植瘤生长有明显的抑制作用,两者联合具有协同增效作用;其机制可能与调控血管生成相关因子HIF-1?、VEGF、VEGFR-2的表达密切相关.     

人参皂苷Rg3和干扰素α抗小鼠血吸虫病肝纤维化效果的比较

目的比较人参皂苷Rg3和IFN-α抗日本血吸虫病肝纤维化的作用。方法以日本血吸虫尾蚴感染ICR小鼠,建立肝纤维化模型,10周后顿服吡喹酮杀虫,以人参皂苷Rg3灌胃治疗和IFN-α皮下注射治疗,肝组织染色,与模型对照组比较,观察和计算各组小鼠肝脏胶原面积百分比及肝纤维化程度,评价人参皂苷Rg3和IFN-α的治疗效果。结果肝纤维化小鼠分别经人参皂苷Rg3和IFN-α治疗8周后,肝脏仍有不同程度增大,暗褐色,质地略韧,肝脏内纤维增生、炎细胞浸润等病变比模型组减轻。Rg3治疗组和IFN-α治疗组小鼠肝脏胶原面积百分比均降低(均P<0.05);SSS评分均低于模型对照组2分以上(均P<0.05),低于实验对照组5分以上(均P<0.05);两治疗组间胶原面积百分比及SSS评分差异无统计学意义(P>0.05)。结论吡喹酮治疗后,人参皂苷Rg3和IFN-α均具有改善日本血吸虫小鼠病肝纤维化作用,因此可以考虑将鲜见毒副作用的人参皂苷Rg3用于血吸虫病肝纤维化的长期治疗。     

以神经内肽酶为靶点治疗阿尔茨海默病的人参皂苷有效组分筛选

目的探讨以神经内肽酶(NEP)作为靶点治疗阿尔茨海默病(AD)的人参皂苷有效组分和筛选方法。方法构建swe APP-SK细胞株作为模型细胞;采用Western印迹及ELISA鉴定构建细胞;采用MTT检测人参皂苷组分对SK-N-SH细胞增殖能力的影响;采用NEP肽酶试剂盒测定人参皂苷组分对NEP肽酶活性的影响;采用ELISA检测筛选有效人参皂苷组分对swe APP-SK细胞株的影响。结果成功构建高表达APP的sweAPP-SK模型细胞,swe APP-SK模型细胞Aβ40、Aβ42表达量均较未转染细胞明显增多,差异均具有统计学意义(P<0.05);人参皂苷组分浓度梯度试验结果显示,各皂苷组分最佳作用浓度分别为皂苷Rg1 20μmol/L,Rg2 50μmol/L,Rg3 50μmol/L,Rb1 50μmol/L,Re 50μmol/L;人参皂苷组分时间梯度试验结果显示,最佳作用时间为48~72 h;人参皂苷Rg1、Rg3、Rb1可明显提高NEP活性,分别为对照组细胞酶活性的(124±3.6)%、(131±4.1)%和(121±3.2)%,差异均具有统计学意义(P<0.05);swe APP-SK模型细胞经Rg1、Rg3、Rb1作用后,细胞外Aβ40、Aβ42浓度均明显降低,差异均有统计学意义(P<0.05)。结论人参皂苷Rg1、Rg3、Rb1可通过NEP做治疗靶点降低swe APP-SK细胞株Aβ40、Aβ42的分泌AD的NEP靶点治疗提供依据。     

一氧化氮合酶抑制剂对小鼠肺癌血管形成的影响

将Lewis肺癌细胞接种于雄性C57小黑鼠前肢腋下,皮下成瘤后,将小鼠随机分成对照组和治疗组,两组分别腹腔注射生理盐水及硝基左旋精氨酸甲基酯(L-NAME),持续14 d.接种第15天检测血清NO含量;处死小鼠,剥离肿瘤称取瘤组织质量、光镜下观察肺内转移灶、测微血管密度(MVD).结果 对照组与治疗组瘤组织质量分别为(2.59±0.31)、(1.80±0.33)g,MVD分别为24.06±5.42、16.97±2.86,NO含量分别为(33.26±12.53)、(14.56±7.14)μmol/L,两组比较均有统计学差异(P均﹤0.05).光镜下观察两组肺内转移情况无明显差异.认为L-NAME对小鼠肺癌血管形成有抑制作用,进而抑制肿瘤生长;Lewis肺癌可能转移较少或出现较晚.     

唑来膦酸对乳腺癌小鼠的抑瘤作用

目的分析唑来膦酸对4T1小鼠乳腺癌种植模型的抑瘤效果。方法选取对数生长期4T1细胞混悬液,将其在32只BALB/c小鼠右腋乳垫区进行接种,细胞悬液测定证明成瘤后,随机将32只小鼠随机平分为A、B、C、D四组,其中A组作为对照,于小鼠腹腔注射0.2 ml生理盐水;B组采用唑来膦酸注射,与每只小鼠皮下注射5μg唑来膦酸;C组采用多西他赛注射,每只小鼠腹腔注射15 mg/kg多西他赛;D组采用唑来膦酸和多西他赛联合注射,首先腹腔注射15 mg/kg多西他赛,然后在同一只小鼠皮下注射5μg唑来膦酸,给药共3 w,应用Pearl Imager TM活体成像系统检查小鼠全身肿瘤病灶及强度;检测计算肿瘤细胞凋亡率及肺脏转移灶数量。结果 D组抑瘤率显著高于A、B、C组(P0.05);C组、D组肺脏转移灶数量显著小于A组、B组(P0.05);D组肿瘤细胞凋亡率明显高于A、B、C组(P0.05)。结论采用唑来膦酸联合多西他赛用药可以对肿瘤生长具有更好地抑制作用、同时可提高细胞凋亡诱导及降低内脏转移效果。     

Surgical approaches of resectable synchronous colorectal liver metastases： Timing considerations

AIM: To compare the safety and efficacy of simultaneous versus two stage resection of primary colorectal tumors and liver metastases. METHODS: From January 1996 to May 2004, 103 colorectal tumor patients presented with synchronous liver metastases. Twenty five underwent simultaneous colorectal and liver surgery and 78 underwent liver surgery 1-3 mo after primary colorectal tumor resection. Data were retrospectively analyzed to assess and compare the morbidity and mortality between the surgical strategies. The two groups were comparable regarding the age and sex distribution, the types of liver resection and stage of primary tumors, as well as the number and size of liver metastases. RESULTS: In two-stage procedures more transfusions were required (4 ± 1.5 vs 2 ± 1.8, pRBCs, P < 0.05). Chest infection was increased after the two-stage approach (26% vs 17%, P < 0.05). The two-stage procedure was also associated with longer hospitalization (20 ± 8 vs 12 ± 6 d, P < 0.05). Five year survival in both groups was similar (28% vs 31%). No hospital mortality occurred in our series. CONCLUSION: Synchronous colorectal liver metastases can be safely treated simultaneously with the primary tumor. Liver resection should be prioritized over colon resection. It is advisable that complex liver resections with marginal liver residual volume should be dealt with at a later stage.     

西罗莫司对人肝癌裸鼠肝脏移植瘤生长的影响

目的 探讨西罗萸司(SRL)对人肝癌裸鼠肝脏移植瘤生长的影响.方法 建立人肝癌裸鼠肝脏移植瘤模型,使用SRL、他克莫司(FK506)进行干预治疗,采用免疫组织化学方法和图像分析技术检测移植瘤血管内皮细胞生长因子、增殖细胞核抗原的表达和微血管密度,原位末端标记法检测肿瘤细胞凋亡情况.统计学处理采用方差分析或t检验.结果 (1)SRL、FK506组和对照组移植瘤质量分别为(352±38)mg、(683±53)mg、(675±45)mg;SRL组移植瘤质量较对照组明显减少(t=10.378,P<0.01);FK506组和对照组比较无明显差异(P>0.05).(2)SRL组移植瘤血管内皮细胞生长因子和增殖细胞核抗原的表达较对照组明显下凋亡(f值分别为5.753和5.296,P<0.05),FK506组和对照组比较无明显差异(P>0.05).(3)SRL组移植瘤微血管密度较对照组明显减少(t=8.637,P<0.01);FK506组和对照组相比无明显变化(P>0.05).(4)SRL组移植瘤凋亡指数明显高于对照组(t=11.518,P<0.05);FK506对移植瘤凋亡指数无明显影响(P>0.05). 结论 SRL可通过减少肿瘤血管形成、阻I卜肿瘤增殖、诱导肿瘤细胞凋亡抑制肝癌的生长.     

Effects of thalidomide on angiogenesis and tumor growth and metastasis of human hepatocellular carcinoma in nude mice

AIM: To investigate the effects of thalidomide on angiogenesis, tumor growth and metastasis of hepatocellular carcinoma in nude mice. METHODS: Twenty-four nude mice were randomly divided into therapy group and control group, 12 mice in each group. Thalidomide dissolved in 0.5% sodium carboxyl methyl cellulose (CMC) suspension was administered intraperitoneally once a day at the dose of 200 mg/kg in therapy group, and an equivalent volume of 0.5% CMC in control group. Mice were sacrificed on the 30th d, tumor size and weight and metastases in liver and lungs were measured. CD34 and VEGF mRNA in tumor tissue were detected by immunohistochemistry and semi-quantitative RT-PCR respectively and microvessel density (MVD) was counted. Serum concentrations of TNF-α and ALT and AFP were also tested. RESULTS: MVD and VEGF mRNA in therapy group were less than those in control group (31.08±16.23 vessels/HP vs 80.00±26.27 vessels/HP, 0.0538±0.0165 vs 0.7373±0.1297, respectively, P<0.05). No statistical difference was observed in tumor size and weight and metastases in liver and lungs. TNF-α was significantly lower in therapy group than in control group (28.64±4.64 ng/L vs 42.69±6.99 ng/L, P<0.05). No statistical difference in ALT and AFP was observed between groups. CONCLUSION: Thalidomide can significantly inhibit angiogenesis and metastasis of hepatocellular carcinoma. It also has inhibitory effects on circulating TNF-α.     

小鼠结肠癌中巨噬细胞金属弹力蛋白酶对血管抑素生成和肿瘤血管生长的影响

目的 在小鼠模型体内探讨巨噬细胞金属弹力蛋白酶(macrophage metalloelastase,MME)分解纤溶酶原生成有生物学功能的血管抑素的途径,及其对肿瘤生长和微血管密度(MVD)的抑制作用.方法 构建重组质粒pEGFP-C1-MME.给90只小鼠分别皮下接种稳定转染pEGFP-C1-MME(MME转染组)、空质粒pEGFP-C1(空质粒转染组)和未转染(未转染组)的CT-26细胞,每组30只.应用放射性碘标纤溶酶原及其示踪技术分析体内血管抑素的生成情况.结果 各组肿瘤标本进行SDS-PAGE电泳,在MME转染组电泳胶中,蛋白相对分子质量为35×103和38×103的区域放射性明显高于空质粒转染组和未转染组(P值均为0.00).Western印迹检测发现三组均检测出35×103和38×103片段表达,通过灰度扫描分析发现上述2个片断在MME转染组表达强度为9.32±1.52和5.61±2.24,明显高于空质粒组(2.47±0.23和0.67±0.12,P值均为0.00)和未转染组(1.21±0.69和0.86±0.44,P值均为0.00).MME转染组MVD平均值和血管内皮生长因子(VEGF)荧光表达强度均明显低于空质粒转染组和未转染组(P值均为0.00).MME转染组小鼠皮下种植瘤平均体积小于空质粒组和未转染组(P值均为0.00).结论 MME是与血管抑素生成密切相关的间质金属蛋白酶,同时具有抑制结肠肿瘤血管形成的作用.

Abstract：

Objective To determine the pathway of macrophage metalloelastase (MME)generate active angiostatin by decomposing plasminogen and its effect on inhibiting growth of tumor and microvessel density (MVD) in vivo in mouse models. Methods The recombined plasmid pEGFPC1-MME was constructed. Thirty mice were subcutaneously inoculated with CT-26 cells that were stably transfected with pEGFP-C1-MME (MME-transfected group), 30 with CT-26 cells transfected with empty vector pEGFP-C1 (vector-transfected group) and 30 with CT-26 cells (non-transfected group). Radioiodination and radioisotope tracer were used to explore the pathway of angiostatin generation in vivo. Results SDS-PAGE electrophoresis analysis revealed that, in the PAGE gel contained the protein with molecular weights of 35 000 and 38 000, radioactivity in MME-transfected group was significantly higher than vector-transfected and non-transfected groups (P = 0. 00).Western blotting analysis demonstrated two bands containing 35 000 and 38 000 fragments in three groups. Quantification of the protein signals by image analysis revealed that the levels of 35 000 and 38 000 fragments were obviously increased in MME-transfected group (9.32±1.52 and 5.61±2.24,respectively) than those in vector-transfected (2.47 ± 0.23 and 0. 67 ± 0. 12, respectively) and nontransfected (1.21±0. 69 and 0. 86 ± 0.44, respectively) groups (P= 0.00). The average value of MVD and fluorescent express of vascular endothelial growth factor (VEGF) were lower in MMEtransfected group when compared with those in vector-transfected and non-transfected groups (P =0.00). The average tumor size in MME-transfected group was small in comparison with vectortransfected and non-transfected groups (P= 0.00). Conclusions MME is demonstrated to be one of matrix metalloproteinase that closely related with angiostatin production and has inhibitory effect on tumor growth in tumor-bearing mice.     

Portal vein ligation and partial hepatectomy differentially influence growth of intrahepatic metastasis and liver regeneration in mice

BACKGROUND/AIMS: Since modern treatment of liver metastases includes serial hepatectomies and portal vein obstruction, we investigated the effects of portal vein ligation (PVL) and partial hepatectomy (PH) on tumor growth. METHODS: The effects of 70% PH and PVL on liver and lung metastases were evaluated in mice. Tumor growth and liver regeneration were assessed by morphometry and immunohistochemistry for PCNA and BrdU. The effect of growth factors of liver regeneration on CT-26 cells was tested in vitro, and TGF-beta secretion of CT-26 cells was measured by ELISA. RNA synthesis of TGF-beta and activin A was measured by RT-PCR. RESULTS: Liver regeneration after PH and PVL was similar in tumor-free mice. Intrahepatic tumor growth was lower after PH than after PVL (p=0.016). Extrahepatic tumor growth was not different. In contrast to PVL, liver regeneration was delayed after PH in metastatic livers (p=0.001). Tested growth factors of liver regeneration stimulated CT-26 cells in vitro, and CT-26 secreted significant amounts of TGF-beta in vitro and in vivo. CONCLUSIONS: Although similar in tumor-free mice, liver regeneration significantly differed between PVL and PH in metastatic livers. In addition, PH and PVL differently affected intrahepatic tumor growth.     

TNP-470 fails to block the onset of angiogenesis and early tumor establishment in an intravital minimal disease model

BACKGROUND AND AIMS: The angiogenesis inhibitor TNP-470 (AGM-1470) has shown encouraging results in animal models of established tumors. However, results of recent clinical trials using TNP-470 have been disappointing. Since little is known about the effects of TNP-470 at the minimal disease stage, we analyzed the effects of TNP-470 on the early stages of tumor establishment. METHODS: Twenty thousand green fluorescent protein (GFP)-transfected murine CT-26 (colonic carcinoma) or Panc-02-H0 (pancreatic adenocarcinoma) cells were inoculated in dorsal skin-fold chambers in BALB/c or C57BL6 mice. Tumor area and microvessel density (MVD) were quantified by intravital microscopy (IVM). Body weight was also monitored. Effects were compared with those in a conventional model involving subcutaneous (s.c.) inoculation of 10(6) tumor cells, followed by measurement of tumor volume, endogenous plasma VEGF/endostatin (ELISA) and proliferation/apoptosis/microvessel density (Ki-67/TUNEL/CD-34). TNP-470 was injected s.c. over the 10-day experimental period (30 mg/kg every 2 days [n=6] to 100 mg/kg/day [n=5 dorsal skin-fold chamber model, n=4 s.c. tumor model]). RESULTS: At 30 mg/kg/every second day neither CT-26 nor PANC-02-H0 tumors were inhibited in neither of the two models. TNP-470 dosage was escalated in CT-26-bearing animals until an antiangiogenic effect could be observed. In the IVM model, only TNP-470 100 mg/kg/day reduced MVD (P=0.006), but failed to block the onset of angiogenesis and tumor area increase. Body weight decreased by 25% (P<0.05). In the subcutaneous tumor model, tumor growth was reduced (P=0.045) but not blocked, while vascular endothelial growth factor (VEGF)/endostatin synthesis and Ki67/TUNEL/CD-34 were not significantly affected. CONCLUSION: While capable of reducing tumor growth in a conventional model, treatment with TNP-470 does not block the onset of angiogenesis and tumor establishment in a model of minimal disease. When used as a single agent TNP-470 does not control minimal tumor disease in experimental colonic carcinoma.     

人参皂甙Rg3抗鼠源性膀胱癌BBT739黏附、侵袭和肺转移作用的研究

目的研究人参皂甙Rg3对鼠源性膀胱癌细胞系BBT739黏附、侵袭和肺转移的作用。方法应用酶标仪检测不同浓度的人参皂甙Rg3对膀胱癌细胞BBT739体外黏附力的影响;通过癌转移模型,观察不同剂量人参皂甙Rg3对膀胱癌小鼠肺转移的影响;体内侵袭模型通过给予不同剂量人参皂甙Rg3观察18d后癌组织的侵袭情况,病理分级检测相关癌的侵袭阶段。结果人参皂甙Rg3可以明显抑制鼠源性膀胱癌细胞系BBT739体外的黏附(P<0.01)及癌细胞对小鼠肾胞膜下的侵袭过程,抑制小鼠膀胱癌肺转移的发展(P<0.05)。结论人参皂甙Rg3通过抑制癌细胞的黏附、侵袭和癌组织抑制癌转移的发生。     

结直肠癌同时性肝转移患者原发灶切除的预后分析

目的 原发灶切除能否使结直肠癌肝转移患者生存获益,目前仍有争议.本研究探讨接受原发灶切除结直肠癌肝转移患者的生存状况及预后的影响因素.方法 回顾性分析2010年1月～2018年2月在国家癌症中心/中国医学科学院肿瘤医院治疗的371例结直肠癌同时性肝转移患者的病例资料.根据治疗方式分为单纯化疗组和原发灶切除组,分析两组患...     

Proliferation of hepatic metastases of colorectal carcinoma: relationship to primary tumours and prognosis after hepatic resection

In this study, we determined the proliferation indices of liver metastatic tumours originating from colorectal carcinomas using Ki67 and argyrophil nucleolar organizer region associated proteins (AgNOR) stain. We examined the primary and metastatic tumours in 27 patients with liver metastasis and eight cases with non-metastatic colorectal carcinoma as a control.The number of AgNOR dots in metastatic tumours was significantly higher than in the respective primary tumours of the metastasis group or in non-metastatic colorectal carcinomas. The Ki67 labelling index was similar in all groups. The Ki67 labelling index and AgNOR counts did not correlate with each other. There was no significant relationship between proliferation indices and the duration of the disease-free period following hepatic resection for metastases or with prognosis after hepatectomy. We conclude that Ki67 and AgNOR are not useful indicators of prognosis in patients who undergo operation for liver metastasis of colorectal carcinomas.     

Characteristic differences between patients who have undergone surgical treatment for lung metastasis or hepatic metastasis from colorectal cancer

OBJECTIVE: The characteristic differences between patients with lung or liver metastases from colorectal carcinoma (CRC) have not yet been clarified. A small group of these patients demonstrate a better prognosis, and the selection criteria for resection of liver and/or lung metastasis are not well defined. It is important to compare and analyze the most common metastatic sites, which include liver metastases and lung metastases. The objective of this study was to compare the characteristics of the two groups in order to identify patients who benefitted from surgical resection of CRC. METHODS: We retrospectively reviewed the medical charts of 80 patients who had undergone resection for liver or lung metastasis from CRC in Fukuoka University Hospital between June 1991 and December 2004. These patients were grouped according to surgical therapy received for the metastases, and separated into two groups, as follows: LUM, lung metastases resection; LIM, liver metastases resection. We evaluated these groups for a set of several factors. RESULTS: The characteristic factors between the two groups (LUM vs. LIM) demonstrated significant differences according to histological differentiation, venous invasion, and lymphatic permeation. There was a statistical difference in the disease-free interval (DFI) between the two groups (947.06 +/- 840.39 days in LUM vs. 246.03 +/- 229.26 days in LIM). Although serum CEA levels at resection of metastasis showed significant differences between the groups (LUM, 13.25 +/- 31.55 ng/ml; LIM, 55.21 +/- 99.52 ng/ml), the primary serum CEA levels were not significantly different. Overall survival rates at 5 years were 37.0 % for LUM and 42.8 % for LIM. There was no significant difference in the survival rate of the LUM vs. the LIM group after resection of metastasis. The Cox proportional hazards regression model was used to determine serum CEA status at the time of the metastases and showed a significant difference indicating poor prognosis for patients with LUM, but the results were not significant for LIM cases. CONCLUSIONS: Candidates for surgical treatment for lung or liver metastases from CRC may be an acceptable for the same valuable approach, even if characteristic differences were observed in each group.