Phase tissue intercellular adhesion molecule-1 expression in nude mice human liver cancer metastasis model

AIM To study the phase cancer tissue intercellular adhesion molecule-1 (ICAM-1) expression of human cancer metastasis model in nude mice, and to analyze the relationship between ICAM-1 expression and the metastasis and recurrence of hepatocellular cancinoma (HCC).METHODS HCC tissues from liver cancer metastasis model in nude mice (LCI-D20) was orthotopically implanted, and ICAM-1 expression in HCC tissues at different growing time were detected by immunodot blot. Tumor size, intrahepatic and extrahepatic metastasis foci were observed by naked eyes and under light microscope.RESULTS ICAM-1 was positively correlated to the tumor growing time (r=0.88, P＜0.01) and tumor size r=0.5, P＜0.05). It was higher in metastatic HCC than in nonmetastatic HCC (8.24±0.95 vs 3.03±0.51, P＜0.01). ICAM-1 content in cancer tissues increased suddenly after metastasis occurred and then maintained in a high level. ICAM-1 was also higher in multimetastasis group than in monometastasis group (10.05±1.17 vs 5.48±0.49, P＜0.05).CONCLUSION Tissue ICAM-1 could predict not only the metastasis of human liver cancer metastasis model in nude mice early and sensitively, but also the metastasis degree. So tissue ICAM-1 may be a potential index indicating the status of metastasis of HCC patients.     

Expression of intercellular adhesive molecule1 in liver cancer tissues and liver cancer metastasis

AIM: To study the relationship between intercellular adhesive molecule-1 (ICAM-1) and liver cancer metastasis and to search for factors to predict metastasis of liver cancer.METHODS: ICAM-1 expression in fresh tissues of normal liver and hepatocellular cancer (HCC) was examined by immunoperoxidase staining. The expression of ICAM-1 in human hepatoma, tumor surrounding tissues and normal livers were semiquantitatively analyzed by Dot immuno blot. Tissue ICAM-1 expression at mRNA level was detected by Northern blot.RESULTS: All 6 cases of normal liver samples were negative in anti-ICAM-1 immunohistochemical staining, 80.0% (36/45) of HCC presented various ICAM-1 expression. The number of positive cells was a little higher in large tumors, tumors with intact capsule and metastasis, but there was no significant difference. Two cases with cancer embolus also had high ICAM-1 expression. ICAM-1 concentration in HCC (13.43 ± 0.09) was higher than that in tumor surrounding tissues (5.89 ± 0.17, P < 0.01) and normal livers (4.27 ± 0.21, P < 0.01). It was also higher in metastasis group (20.24 ± 0.30) than in nonmetastasis group (10.23 ± 0.12, P < 0.05). Northern blot analysis revealed that ICAM-1 expression at mRNA level was also higher in HCC and cancer embolus than that in tumor surrounding tissues and normal livers.CONCLUSION: Tissue ICAM-1 could indicate the growth and metastasis of HCC, and may be an index that can predict liver cancer metastasis.     

细胞间粘附分子-1在肝癌组织的表达及意义

目的：研究细胞间粘附分子－１（ＩＣＡＭ－１）在原发性肝癌的临床意义。方法：以免疫线化方法结合全自动图像分析检测ＩＣＡＭ－１在３０例原发性肝癌组织及６例正常肝组织的表达,分析其与肿瘤分化程度、大小及转移之间的关系。结果：３０例原发性肝癌组织２５例ＩＣＡＭ－１表达阳性（阳性率８３．３％）,肝癌组织中ＩＣＡＭ－１含量明显高于正常肝组织（Ｐ〈０．０１）,转移组肝癌中ＩＣＡＭ－１含量明显地无转移组（Ｐ〈０．     

Invasion and metastasis of liver cancer: expression of intercellular adhesion molecule 1

To study the relationship between intercellular adhesion molecule 1 (ICAM-1) and liver cancer metastasis and to find predicting factors that could indicate the growth and metastasis of liver cancer. Methods: ICAM-1 expression in fresh tissue of normal liver and hepatocellular cancer (HCC) was examined by immunoperoxidase staining. Serum soluble intercellular adhesion molecule 1 (sICAM-1) from patients with a benign HCC tumor, and the expression of ICAM-1 in the orthotopically transplanted LCI-D20 tumor of a nude mouse liver cancer metastasis model, and in human hepatoma, the tumor surrounding tissue and normal liver, was analyzed semiquantitatively by the immuno-dot blot method. Tissue ICAM-1 expression (mRNA level) was detected by Northern blotting. Results: ICAM-1 expression in LD1-20 D metastatic liver cancer had a positive correlation with tumor size and the time after implantation. It increased suddenly as metastasis occurred being 3.03 ± 0.51 before metastasis and 8.24 ± 0.95 after metastasis, P < 0.01, then remained high, appending on the number of sites involved (monosite metastasis 5.48 ± 0.49, multisite metastasis 10.05 ± 1.17, P < 0.05). All six cases of normal liver samples were negative in anti-ICAM-1 immunohistochemical staining, 80.0% (36/45) of the HCC showed some ICAM-1 expression. The rate of positive cells was a little higher in large tumors, tumors with an intact capsule and tumors with metastasis, but there was no significant difference. It was noticed that two cancer emboli also had high ICAM-1 expression. The ICAM-1 concentration in HCC (13.43 ± 0.09) was higher than that in tumor surrounding the liver (5.89 ± 0.17, P < 0.01) and that in normal liver (4.27 ± 0.21, P < 0.01). sICAM-1, like tissue ICAM-1, was higher in HCC patients than in patients (with benign liver tumor and normal controls. Both tissue ICAM-1 and sICAM-1 were higher in the metastasis group than in the group without metastasis (tissue ICAM-1 20.24 ± 0.30 vs 10.23 ± 0.12 P < 0.05; sICAM-1 12.18 ± 0.25 vs 9.77 ± 0.54 P < 0.05). Northern blot analysis revealed that ICAM-1 expression, as indicated by mRNA level, was also higher in HCC and in cancer emboli than in tumor surrounding liver and normal liver. Conclusions: Tissue ICAM-1 and serum sICAM-1 could indicate the stage of HCC, and the potential of hepatoma cells for invasion and metastasis. They may play an important role in the metastasis cascade. Received: 20 January 1998 / Accepted: 25 June 1998     

敲除NOR1基因对人肝癌裸鼠移植瘤的影响及作用机制

目的 探究敲除NOR1基因后对人肝癌裸鼠移植瘤生长、存活率及器官损伤的影响。 方法 SPF级雄性BALB/C裸鼠30 只随机分为Control组(n=10)、Scramble组(n=10)、siNOR1组(n=10),构建阴性对照质粒 (Scramble)和 siNOR1质粒转染至HepG2细胞并接种至裸鼠构建移植瘤裸鼠模型。连续30 d每5 d检测裸鼠移植瘤组织体积;检测移植瘤重量;免疫组化检测Ki-67、Caspase-3、Notch、NOR1表达情况;Western Blot检测Survivin、Notch1、NICD、Hes1、Hey1蛋白表达量情况,HE染色观察肝、肾、肺、脑病理损伤程度。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验;生存分析采用Kaplan-Meier法,生存率的比较采用log-rank检验法。结果 从建模第20天开始,与Control组相比,siNOR1组第20、25、30 天移植瘤体积显著减小[(418.71±78.24)mm3 vs (149.6.16±60.05)mm3、(864.22±125.66)mm3 vs (239.83±100.51)mm3、(1468.45±199.78) mm3 vs (446.54±147.09) mm3,P值均<0.05];第30天时取出移植瘤,与Control组相比,siNOR1组移植瘤质量显著减小[(0.45±0.07)g vs (0.12±0.04)g,P<0.05];与Control组相比,siNOR1组移植瘤组织中Ki-67、Notch、NOR1阳性表达率显著降低[Ki-67:(48.98±9.75)% vs (11.51±5.09)%, Notch: (62.51±9.26)% vs (18.75±4.61)%,NOR1:(76.33±8.31)% vs (16.57±3.76)%, P值均<0.05],Caspase-3阳性表达率显著升高[(6.39±4.67)% vs (38.03±9.28)%, P<0.05];siNOR1组模型裸鼠30 d内生存率显著高于Control组[(77.66±6.75)% vs (25.32±4.63)%,χ^2=6.897,P<0.05];Western Blot检测结果显示,与Control组相比,siNOR1组Survivin、Notch1、NICD、Hes1、Hey1蛋白表达水平均显著降低(Survivin: 0.34±0.06 vs 0.02±0.01;Notch1:0.16±0.03 vs 0.03±0.01;NICD:0.26±0.05 vs 0.04±0.02;Hes1:0.35±0.04 vs 0.06±0.02;Hey: 0.29±0.06 vs 0.05±0.02,P值均<0.05),且siNOR1组裸鼠各组织器官损伤情况均得到有效缓解。结论 敲除NOR1基因抑制人肝癌裸鼠移植瘤的生长,提高模型裸鼠生存率,并缓解模型裸鼠肝、肾、肺、脑损伤,其机制可能与抑制Notch信号通路活性有关。     

原发性肝细胞癌中WAF1／CIP1基因的及其与p53基因突变的关系

目的 研究原发性肝细胞癌（ＨＣＣ）组织中ＷＡＦ１基因的表达及其与ｐ５３基因突变和临床病理学指标的关系和意义。方法 应用半定量ＲＴ－ＰＣＲ、免疫组织化学及定量ＤＮＡ图像分析的方法,检测ＷＡＦ１基因在３２例ＨＣＣ及其癌旁肝组织和５例正常肝细胞中的表达,研究其与ｐ５３基因突变及临床病理学指标的关系。结果 肝癌组织中ＷＡＦ１ｍＲＮＡ表达相对值（１．０６±０．３７）Ｕ低于其相应旁旁肝组织（１．３０±０．３７     

锌α2糖蛋白对实验性大鼠肝癌的抑制作用

目的探讨锌α2糖蛋白(AZhGP1)与肝细胞癌(HCC)发生、发展和转移的关系。方法二乙基亚硝胺(DEN)构建肝硬化和HCC大鼠模型,以过表达AZGP1基因干扰的HepG2细胞液构建裸鼠皮下成瘤和肝原位移植模型。免疫组织化学、蛋白免疫和PCR检测AZGP1和TGFβ1的表达。结果AZGP1 mRNA在正常肝组织、肝硬化和肝癌中的表达分别为(0.98±0.02)、(0.52±0.03)、(0.20±0.02);而TGFβ1的基因和蛋白表达在肝硬化和肝癌组织中呈现显著上调。AZGP1过表达的HepG2细胞接种裸鼠构建皮下肿瘤(HepG2-AZGP1组)。与对照组(HepG2-GFP组)比较,肿瘤大小无差异。裸鼠肝内移植术6周后,HepG2-GFP组57%和HepG2-AZGP1组14%发生肺转移(P=0.0157)。与HepG2-GFP组比较,HepG2-AZGP1组肝脏原发灶和肺转移灶结节数目明显减少,癌细胞异型性明显较轻。结论肝硬化进展至肝癌过程中抑癌基因.AZGP1发生缺失,伴随着失去阻抑TGFβ1作用。恢复AZGP1功能可能是一种新的有前途的治疗肝癌方法。     

Study on the inhibitory effect of tyroserleutide on tumor growth and metastasis in nude mice model of human hepatocellular carcinoma metastasis

BACKGROUND/AIMS: Previous study has shown that small tripeptide tyroserleutide has inhibitory effect on liver cancer both in vitro and in vivo. This study was designed to test the effect of tyroserleutide on tumor growth and metastasis in a nude mice model of human hepatocellular carcinoma (HCC) metastasis. METHODOLOGY: Highly metastatic human HCC cell HCCLM6 was used to construct orthotopic implantation model of HCC metastasis in 20 BALB/c-nu/nu nude mice, which were randomized into treatment group and control group each with 10 mice. The former received daily CMS024 intraperitoneal injection at the dose of 300 microg/kg beginning from the second postoperative day, and the latter received intraperitoneal injection of equal amount of 0.85% sodium chloride solution. The mice were observed for signs of disease development. Thirty-five days later, the mice were sacrificed and abdominal and pulmonary metastasis was recorded and peripheral blood hematological and biochemical parameters were determined. RESULTS: After 35 days of intervention, all the 10 mice in the treatment group were alive and well, but there were only 9 living mice in the control group. The mean tumor weights were 1.9 +/- 0.5 g in the treatment group and 2.3 +/- 0.8 g in the control group (P > 0.05). The abdominal wall metastasis and intraperitoneal metastasis were 100% for the control group and only 60% and 50%, respectively, for the treatment group (P < 0.05). Bloody ascites and gross intrahepatic metastatic nodules were found in 70% and 90% of nude mice, respectively for the control group, and 20% and 40%, respectively, for the treatment group (P > 0.05). The median number of grades I, II, III and IV pulmonary metastasis were 92, 24, 15 and 16, respectively, for the control group, and 24, 20, 10 and 8, respectively, for the treatment group. There were statistically significant differences in the number of grade I pulmonary metastasis between the treatment group and the control group (24 vs. 92, P < 0.05). No statistically significant differences in the body weight, peripheral blood hematological and biochemical parameters were observed between the two groups. No toxic effects were observed during the observation period. CONCLUSIONS: Small molecule tyroserleutide demonstrated positive effects to retard tumor growth and inhibit loco-regional and long distance metastasis in a nude mice model of human HCC metastasis, and no obvious toxic or side effects on the tumor bearing nude mice.     

巨噬细胞移动抑制因子和细胞周期蛋白D1的表达与肝细胞癌生长和转移的关系

目的研究巨噬细胞移动抑制因子（MIF）、细胞周期蛋白D1（Cyclin D1）、细胞周期蛋白依赖激酶4（CDK4）、磷酸化视网膜母细胞瘤易感基因产物蛋白（phospho—Rb）在HeC组织中的表达及其与癌细胞生长和转移的关系。方法应用组织芯片技术和免疫组织化学方法检测93份HCC组织中MIF、Cyclin D1、CDK4和phospho—Rb的表达，分析它们的表达与HCC临床病理特征的关系。结果93份HCC组织中MIF、Cyclin D1、CDK4和phospho—Rb的表达率分别为71％、41％、82％和14％，MIF和Cyclin D1阳性表达率与正常肝脏组织表达率之间的差异有统计学意义（P〈0．01），CDK4和phospho—Rb阳性表达率与正常肝脏组织表达率之间的差异无统计学意义（P〉0．05）。在≥3．5cm的肿瘤中MIF表达率为79％，明显高于在〈3．5cm肿瘤中的表达率48％（P〈0．01）；有转移的肝癌组织中Cyclin D1阳性率为62％，明显高于无转移组织中的阳性率35％，差异有统计学意义（P〈0．05）。MIF表达强弱与Cyclin D1的表达呈正相关关系（P〈0，01）。CDK4和phospho—Rb的表达与肿瘤大小及是否转移的关系无统计学意义。结论MIF和Cyclin D1在HCC发展进程中可能促进肿瘤的生长和转移。     

Quantitation of alpha-fetoprotein messenger RNA in peripheral blood of nude mice and its relationship with tumor recurrence and metastasis after curative resection of hepatocellular carcinoma]

OBJECTIVE: To assess the level of alpha-fetoprotein (AFP) messenger RNA (mRNA) in peripheral blood of nude mice, and to study its relationship with tumor recurrence and metastasis after curative resection of hepatocellular carcinoma (HCC). METHODS: The metastatic model of human HCC in nude mice LCI-D20 was used in this study. Curative resection was performed at 10th day after tumor implantation in 28 nude mice. Interferon alpha-1b (IFN alpha-1b) was administered subcutaneously from the next day after resection, at doses of 3 10(7)U/kg/d (8 nude mice), 1.5 10(7) U/kg/d (8 nude mice) respectively in the treatment groups, and normal saline alone in the controlled group (12 nude mice). Thirty-five days after treatment, one milliliter of peripheral blood was taken and AFP mRNA was quantitatively analyzed using TaqMan real-time quantitative RT-PCR. The mice were sacrificed. The size of recurrent tumor was measured and the presence of lung metastases was observed. RESULTS: The liver recurrent rate, lung metastatic rate and positivity of AFP mRNA in the controlled group were all 100% (12/12), whereas it was 62.5% (5/8), 0% (0/8), 87.5% (7/8) respectively in the IFN alpha-1b 1.5 10(7)U/kg/d treated group. The recurrent tumor in liver of the IFN alpha-1b 1.5 10(7)U/kg/d treated group was much smaller than that of the controlled group (25 mm(3) 2 mm(3) vs 1143 mm(3) 3 mm(3), t =9.27, P<0.01), and the level of AFP mRNA was also lower than that of the controlled group [(85 6)copies/mug vs (955 2) copies/mug, t =4.33, P<0.01]. In the IFN alpha-1b 3 10(7)U/kg/d treated group, there was only one recurrent tumor (0.5 mm(3)), no lung metastasis, and the positivity of AFP mRNA was 0% (?(2)=11.67, P<0.01). CONCLUSIONS: These results suggest that the level of AFP mRNA in peripheral blood may indicate recurrence and/or metastasis after curative resection of HCC. TaqMan real time quantitative RT-PCR is a very sensitive and convenient method for detecting circulating cancer cells.     

达肝素钠对肝癌生长转移抑制的实验研究

目的研究低分子肝素达肝素钠对肝癌生长转移的抑制作用。方法采用人肝癌裸鼠转移模型(LCI-D20)。40只模型鼠随机分成4组即对照组、化疗组(顺铂 氟尿嘧啶),达肝素钠组、联合组(顺铂、氟尿嘧啶与达肝素钠)。观察肿瘤大小和转移、抑瘤率、测血清甲胎蛋白(AFP)、肿瘤微血管密度(MVD)、CD31。结果对照组、化疗组、达肝素钠组、联合组的肿瘤体积分别为(25245±13367)mm3、(1610 ±1217)mm3、(5883±3131)mm3和(5556±2570)mm3;抑瘤率分别为0%、93.6%、76.7%和78.0%;MVD 分别为20.7±6.8、18.2±2.6、4.8±1.8和6.5±2.4;CD31分别为31.8±5.7、25.5±5.1、21.6±4.8和19.6±2.4;AFP分别为(121.8±31.4)ng/ml、(21.5±13.3)ng/ml、(75.6±29.7)ng/ml 和(55.8±38.0)mg/ml;肝转移率分别为80%、70%、20%和10%;肺转移率分别为70%、60%、20%和10%; 腹壁转移率分别为90%、60%、30%和30%;腹水形成率分别为20%、10%、0%和0%。化疗组、达肝素钠组、联合组分别与对照组比,对肝癌生长的抑制作用差异有统计学意义,F=9.191,P<0.01。达肝素钠对肝癌血管形成和转移有良好的抑制作用,与对照组及单纯化疗组比差异有统计学意义,F=4.937,P<0.01。结论低分子肝素达肝素钠通过抗肿瘤血管形成,对肝癌的生长与转移有抑制作用。     

Tiam1、Rac1在评估大肠癌远隔脏器转移中的意义

目的:观察人大肠癌和癌旁正常大肠黏膜组织中T淋巴瘤侵袭转移诱导因子1(Tiam1)和RacGTP酶激活蛋白1(Rac1)的表达,并探讨其与大肠癌根治术后发生远隔脏器转移的相关性.方法:应用SP免疫组织化学法检测87例大肠癌及40例癌旁正常组织标本中Tiam1/Rac1的表达情况;应用竞争性RT-PCR检测上述标本中Tiam1 mRNA的表达情况;应用配体沉淀法检测上述标本中Rac1蛋白活性.结果:Tiam1/Rac1在大肠癌细胞胞质内呈阳性染色.Tiam1 mRNA在大肠癌中表达明显增强(0.6±0.02vs0.24±0.02,P<0.0005),并且在术后发生远隔转移患者中的表达要高于未发生远隔转移者(0.91±0.02vs0.52±0.02,P<0.0005).Rac1蛋白活性的表达情况与Tiam1一致,在大肠癌中表达高于正常黏膜(0.17±0.01vs0.07±0.05,P<0.0005),并且在有远隔转移的大肠癌组织中活性要高于无远隔转移者(0.25±0.02vs0.15±0.01,P<0.0005).对于不同分化程度的大肠癌组织,Tiam1/Racl的含量均没有统计学差异(0.63±0.04,0.60±0.04,0.57±0.04,P=0.613;0.18±0.06,0.17±0.05,0.15±0.05,P=0.558).结论:Tiam1/Racl是大肠癌根治术后患者发生远隔转移的正性因子,可以作为预测大肠癌根治术后患者发生远隔脏器转移的有效指标.     

NDRG1在原发性肝细胞癌及胎肝中的表达及意义

目的:探讨NDRG1在原发性肝细胞癌(HCC)及胎肝组织中的表达及其意义.方法:收集2002-01/2008-12广州市第一人民医院手术切除的肝细胞癌标本81例. 所有患者术前未行放疗和化疗; 25例胎肝组织, 取自不同月份流产或引产的胎儿(4、5、6、7、8 mo胎儿各5例); 另选43例癌旁组织, 10例肝硬化组织, 9例正常肝组织(移植肝), 8例原发癌转移灶组织作为对照. 观察肝脏组织病理形态特征, 并用免疫组织化学EnVison法检测NDRG1的表达.结果:NDRG1在正常肝组织中呈强阳性表达, 平均吸光度值为0.206±0.056, 随着肿瘤的发生, 在癌旁组织中有减弱(0.176±0.083),在HCC中表达明显减弱(0.128±0.096), 在转移灶中表达最低(0.059±0.051), 而在胎肝组织中表达亦较低(0.059±0.074). 各组总体差异均有统计学意义(F = 33.669, P <0.05). HCC与患者年龄、性别、肝炎病史、肝硬化、肿瘤大小、AFP值、HbsAg、淋巴结转移及有无远处转移、肿瘤分型、Child-Pugh分级、TNM分期、CLIP分期均无关(P >0.05), 但与肿瘤的Edmondson分级有关(F = 2.881, P <0.05).结论:NDRG1在HCC中低表达, 并且随着肿瘤的发生发展, 表达量逐渐降低. NDRG1可能对HCC起着抑制作用, 提示该基因可望成为早期预测肝癌转移的分子生物学标志物之一.     

Inhibitory effects of synthetic β peptide on invasion and metastasis of liver cancer

Purpose: To study the inhibitory effects of synthetic β peptide on invasion and metastasis of liver cancer. Methods: Membrane-type intercellular adhesion molecule-1 (ICAM-1) expression of SMMC-7721 cultured hepatoma cells (7721 cells) was detected by immunofluorescence cell flowmeter. The adhesion of 7721 cells to fibronectin (FN) was assayed by the MTT method. The adhesion of 7721 cells to 7721 cells, 7721 cells to endothelial cells, and 7721 cells to lymphocyte cells was detected by adhesion assay. LCI-D20 human liver cancer metastasis model in nude mice was used in this experiment. One hundred micrograms of β peptide per mouse were injected subcutaneously after tumor was resected premetastatically or postmetastatically to observe its effect on liver cancer metastasis after hepatectomy. Results: Membrane-type ICAM-1 expression of SMMC-7721 cells treated by β peptide was lower than that of the untreated cells. The adhesion of 7721 cells to FN, 7721 cells to 7721 cells, 7721 cells to endothelial cells, and 7721 cells to lymphocyte cells was also lower in the β peptide group than in the untreated group. Conclusions:β Peptide can block the adhesion of 7721 cells to FN, 7721 cells to some host cells in vitro, and inhibit HCC metastasis of LCI-D20 model posthepatectomy in vivo, so it could potentially act as an anti-metastasis drug. Received: 23 November 1999 / Accepted: 10 March 2000     

膜型基质金属蛋白酶-1与肝细胞癌侵袭转移性的关系

目的 探索膜型基质金属蛋白酶－１（ｍｅｍｂｒａｎｅ－ｔｙｐｅ１ｍａｔｒｉｘ ｍｅｔａｌｌｏｐｒｏｔｅｉｎａｓｅ,ＭＴ１－ＭＭＰ）与肝细胞癌（ＨＣＣ）侵袭转移性的关系及以ＭＴ１－ＭＭＰ来判断肝细胞癌侵袭转移性的可能性和方法。方法 通过ＲＴ－ＰＣＲ对正常肝、ＨＣＣ及其癌旁组织、癌栓组织,裸鼠人肝癌高、低转移模型中的ＭＴ１－ＭＭＰｍＲＮＡ进行半定量研究,并与ＨＣＣ侵龚转移的病理指标进行统计分析,结果 正     

Metastatic human hepatocellular carcinoma models in nude mice and cell line with metastatic potential

Metastatic human HCC model is needed for the studies onmechanism and intervention of metastatic recurrence,Byusing orthotopic implantation of histologically intacttissues of 30 surgical specimens,a patient-likemetastatic model of human HCC in nude mice (LCI-D20)and a low metastatic model of human HCC in nude mice(LCI-D35) have been established.All mice withtransplanted LCI-D20 tumors exhibited extremely highmetastatic ability including spontaneous metastasis toliver,lungs,lymph nodes and peritoneal seeding.Remarkable difference was also found in expression ofsome of the invasiveness related genes and growthfactors between the LCI-D20 and LCI-D35 tumors.PAI-1increased gradually following tumor progression in LCI-D20 model,and correlated with tumor size and AFP level.Phasic expression of tissue intercellular adhesionmolecule-1 in this model was also observed.Using cornealmicropocket model,it was demonstrated that the vascularresponse induced by LCI-D20 tumor was stronger than thatinduced by LCI-D35 tumor.Similar report on metastatichuman HCC model in nude mice and human HCC cell linewith metastatic potential was rarely found in theliterature.This LCI-D20 model has been widely used forthe studies on intervention of metastasis,including anti-angiogenesis,antisense approach,metalloproteinaseinhibitor,differentiation inducer,etc.It is concluded thatthe establishment of metastatic human HCC model in nudemice and human HCC cell line with metastatic potentialwill provide important models for the in vivo and in vitrostudy of HCC invasiveness,angiogenesis as well asintervention of HCC recurrence.     

Effects of Linomide on growth and metastasis of implanted human gastric cancer in nude mice

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Auphen and dibutyryl cAMP suppress growth of hepatocellular carcinoma by regulating expression of aquaporins 3 and 9 in vivo

AIM: To investigate whether the regulation of aquaporin 3(AQP3) and AQP9 induced by Auphen and dibutyryl c AMP(dbc AMP) inhibits hepatic tumorigenesis. METHODS: Expression of AQP3 and AQP9 was detected by Western blot, immunohistochemistry(IHC), and RT-PCR in HCC samples and paired non-cancerous liver tissue samples from 30 hepatocellular carcinoma(HCC) patients. A xenograft tumor model was used in vivo. Nine nude mice were divided into control, Auphen-treated, and dbc AMP-treated groups(n = 3 for each group). AQP3 and AQP9 protein expression after induction of xenograft tumors was detected by IHC and m RNA by RT-PCR analysis. The terminal deoxynucleotidyl transferase-mediated d UTP nick end labeling assay and histological evaluation were used to detect apoptosis of tumor cells, and the concentration of serum α-fetoprotein(AFP) was measured using RTPCR and an ELISA kit.RESULTS: The volumes and weights of tumors decreased significantly in the Auphen- and dbc AMP-treated mice compared with the control mice(P 0.01). The levels of AQP3 were significantly lower in the Auphen treatment group, and levels of AQP9 were significantly higher in thedbc AMP treatment mice than in the control mice(P 0.01). The reduction of AQP3 by Auphen and increase of AQP9 by dbc AMP in nude mice suppressed tumor growth of HCC, which resulted in reduced AFP levels in serum and tissues, and apoptosis of tumor cells in the Auphen- and dbc AMP-treated mice, when compared with control mice(P 0.01). Compared with para-carcinoma tissues, AQP3 expression increased in tumor tissues whereas the expression of AQP9 decreased. By correlating clinicopathological and expression levels, we demonstrated that the expression of AQP3 and AQP9 was correlated with clinical progression of HCC and disease outcomes. CONCLUSION: AQP3 increases in HCC while AQP9 decreases. Regulation of AQP3 and AQP9 expression by Auphen and dbc AMP inhibits the development and growth of HCC.