健脾理气汤对裸鼠人肝癌高转移模型的影响

通过健脾理气汤对肝癌高转移模型加脾虚模型作用的研究，观察其对肝癌转移的影响，探讨其对肝癌转移干预的机制。     

重组人内皮抑素腺病毒抑制肝癌裸鼠移植瘤生长

目的 观察重组人内皮抑素腺病毒(Ad/hEndo)对人肝癌裸鼠移植瘤生长的影响。方法 人脐静脉内皮细胞ECV-304经Ad/hEndo感染后,western印迹检测人内皮抑素的表达。人肝癌BEL-7402细胞移植到裸鼠背脊部后,检测Ad/hEndo对肝癌移植瘤生长的抑制作用。逆转录聚合酶链反应(RT-PCR)检测肿瘤组织中内皮抑素mRNA的表达。分析人内皮抑素在裸鼠体内的表达分布。结果 Western印迹检测到人内皮抑素基因在ECV-304细胞内高效表达。Ad/hEndo明显抑制人肝癌BEL-7402裸鼠移植瘤生长(F=4.061,P<0.05)。Ad/hEndo组血管密度计数为6.88±1.08,DMEM组为13.60±1.71(t=9.216,P<0.01)。瘤内注射Ad/hEndo后3d,RT-PCR在肿瘤组织检测到内皮抑素mRNA的表达,7d后表达不明显。人内皮抑素蛋白主要分布在肿瘤组织。结论 腺病毒介导的人内皮抑素基因在体内、体外获得高效表达,并明显抑制肝癌裸鼠移植瘤的生长与血管生成。     

抗端粒酶核酶抑制肝癌细胞裸鼠移植瘤生长的研究

设计针对端粒酶RNA组分的特异性核酶，研究该核酶对肝癌细胞裸鼠移植瘤生长的影响。构建含针对端粒酶RNA组分锤头状核酶基因的重组真核表达质粒pBBS212-RZ，以及建立人肝癌细胞株HepG2裸鼠移植瘤模型。将不同剂量的pBBS212-RZ用脂质体包裹后进行瘤体内多点注射，对照组注射生理盐水或空质粒载体pBBS212。连续注射2l天后，测量移植瘤体积和重量，检测瘤组织端粒酶活性。抗端粒酶特异性核酶使瘤组织端粒酶活性下降(抑制率为72％)，明显地抑制了肝癌细胞裸鼠移植瘤生长(抑制率为68％)，且存在剂量依赖性。抗端粒酶特异性核酶作为一种有效的端粒酶抑制剂，可抑制肝癌细胞的生长，有望在肝癌基因治疗中发挥作用。     

肝癌细胞不同制悬方法建立裸鼠移植瘤模型的比较

目的比较肝癌细胞不同制悬方法建立裸鼠皮下瘤及原位瘤模型,优选建模方法。方法将肝癌细胞分别采用PBS缓冲液(PBS组)、无血清DMEM溶液(DMEM组)和含10%血清的DMEM溶液(血清DMEM组)3种液体制悬后注入裸鼠皮下建立肝癌裸鼠皮下瘤模型,比较皮下瘤的体积、瘤重及成瘤率;将肝癌细胞分别采用PBS缓冲液(PBS组)、无血清DMEM溶液(DMEM组)和含10%血清的DMEM溶液(血清DMEM组)3种液体制悬后注入裸鼠肝脏建立肝癌裸鼠原位瘤模型,比较原位瘤的成瘤率。结果3组裸鼠皮下瘤模型的皮下瘤体积及瘤重相比,差异无统计学意义(P0.05),但是PBS组及DMEM组的成瘤率(90%和100%)明显高于血清DMEM组(40%),差异有统计学意义(P0.05);裸鼠原位瘤模型中PBS组的成瘤率(90%)明显高于DMEM组(40%)及血清DMEM组(20%),差异有统计学意义(P0.05)。结论采用PBS缓冲液或无血清DMEM溶液对肝癌细胞稀释制悬可以有效建立裸鼠皮下瘤模型,采用PBS缓冲液对肝癌细胞稀释制悬可以有效建立裸鼠原位瘤模型。     

微囊化人胰腺癌细胞建立裸鼠胰腺癌模型研究

目的研究微囊化人胰腺癌细胞建立裸鼠胰腺癌模型效果,以期建立稳定的更为理想的胰腺癌动物模型.方法 分别以人胰腺癌细胞悬液和微囊化人胰腺癌细胞悬液建立皮下移植瘤模型,定期监测皮下肿瘤大小,绘制生长曲线并进行比较.分别以人胰腺癌细胞悬液和微囊化人胰腺癌细胞建立原位移植瘤模型,分别于模型建立后第4周和第8周进行正电子发射计算机...     

人胃癌裸鼠原位移植模型的生物学行为研究

目的建立人胃癌裸鼠胃壁原位移植瘤模型，并与相应的皮下移植瘤作比较，以探讨宿主器官微环境对胃癌细胞浸润及转移等生物学行为的影响．方法将人胃癌细胞系ＭＧｃ８０３及其克隆株Ｃ１１癌细胞分别接种于裸鼠胃壁及背部皮下，比较原位和皮下移植瘤的成瘤率、生长率、生长方式及浸润、转移等生物学行为，以及体外回复培养后瘤细胞的增殖能力．结果胃壁原位及皮下移植瘤体内成瘤率、生长率及形态学上均无明显不同；其体外增殖能力也无显著性差异．但皮下移植瘤多呈局限性生长，无肝、脾、肾转移，其转移仅限于肺及个别局部淋巴结．胃壁原位移植瘤则浸润破坏胃壁各层组织结构，并直接蔓延到邻近各器官组织．其转移既有经血道至肝、肺、脾、肾等部位，也有经淋巴道至多数局部及远处淋巴结，其淋巴结的转移率较皮下移植瘤有显著增高（Ｐ＜００５）；且多伴有腹、盆腔内广泛种植性转移．结论裸鼠胃壁微环境较皮下组织更适合于人胃癌ＭＧｃ８０３及Ｃ１１移植瘤的浸润及转移的表达，该原位移植瘤模型的恶性生物学行为更接近临床胃癌患者的体内侵袭及转移的实际．     

Survivin反义寡核苷酸抑制肝癌移植瘤生长的作用

目的:检测Survivin反义寡核苷酸(ASODN)在人肝癌耐药细胞株裸鼠皮下移植瘤中的表达情况．方法:将30只裸鼠建立人肝癌耐药细胞系 SMMC-7721／ADM皮下移植瘤模型,随机分成6组:空白对照组(A)、脂质体转染对照组 (B)、正义链对照组(C)、200(D)、400(E)和 600 μg/L(F)反义链组(ASODN组),用不同的转染液注射后2,4,8,12,16,20 d,用逆转录聚合酶链反应技术(RT-PCR)和Western blot蛋白免疫印迹法检测治疗后各组肿瘤组织中Survivin mRNA和蛋白表达的变化．结果:注射后20 d,ASODN组肿瘤细胞生长明显抑制,空白对照组、脂质体转染对照组和正义链对照组裸鼠的mRNA和蛋白表达无明显差异,而ASODN组mRNA和蛋白表达随着时间和浓度的增加,Survivin表达减弱,E,F组与其余个组(A,B,C,D)相比有显著差异(mRNA: 0．33±0．04,0．28±0．03 vs 0．82±0．02,0．78± 0．05,0．72±0．04,0．57±0．03,P<0．05;蛋白: 34．9±3．89,21．2±3．65 vs 72．14±6．53,69．31 ±5.34,68．29±4．98,53．8±5．23,P<0．05)．结论:Survivin反义寡核苷酸能够下调 Survivin mRNA和蛋白的表达,抑制裸鼠皮下移植瘤的生长．     

Anti-tumor activities and apoptosis-regulated mechanisms of bufalin on the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice

AIM： To investigate anti-tumor activities and apoptosisregulated mechanisms of bufalin in the orthotopic transplantation tumor model of human hepatocellular carcinoma in nude mice.
METHODS： BEL-7402 cells of human hepatocellular carcinoma were inoculated to form subcutaneous tumors, and were implanted into the liver to establish orthotopic transplantation tumor models of human hepatocellular carcinoma in nude mice. Seventy-five animals were randomized divided into five groups （n = 15）. Bufalin was injected intraperitoneally into three groups at doses of 1.5 mg/kg （BF1）, 1 mg/kg （BF2） and 0.5 mg/kg （BF3） for d 15-24, respectively. The NS group was injected an equal volume of saline as above and adriamycin was injected intraperitoneally into the ADM group at a dose of 8.0 mg/kg for d 15. Ten mice in each group were killed at d 25 and the survival time in each group was calculated. We also observed the morphologic alterations in the myocardium, brain, liver, kidney and tumor tissues by pathology and electron microscopy, measured the apoptotic rate by TUNEL staining method, and detected the expression of apoptosis-regulated genes bcl-2 and bax by immunohistochemical staining and RT-PCR in tumor tissues.
RESULTS： The tumor volumes in each group of bufalin were reduced significantly （35.21±12.51 vs 170.39 ± 25.29; 49.83 ± 11.46 vs 170.39 ± 25.29; 83.99 ± 24.63 vs 170.39 ± 25.29, P 〈 0.01, respectively）, and the survival times were prolonged in group BF1-2 （31.8 ± 4.2 vs 23.4 ± 2.1 and 29.4 ± 3.4 vs 23.4 ± 2.1, P 〈 0.05, respectively）, and necrosis was mainly in severe or moderate degree in group BF1-2. No morphological changes were detected in the myocardium, brain, liver and kidney tissues. Apoptotic characteristics could be seen in group BF1-2. The positive rates of bcl-2 and bax protein expression of each group by immunohistochemical staining were 10.0%, 10.0%, 20.0%, 10.0% and 20.0%; 90.0%, 80.0%, 80.0%, 40.0% and 30.0%, respectively. Loss of expression of bcl-2 mRNA     

Metastatic models of human liver cancer in nude mice orthotopically constructed by using histologically intact patient specimens

In this study of orthotopic implantation of histologically intact surgical specimens, the authors constructed metastatic models of human hepatocellular carcinoma (HCC) in nude mice. Histologically intact human liver cancer specimens, derived from patients, were implanted directly into the liver of nude mice, and their orthotopic growth and metastases were observed. The transplantability and metastatic rate of two specimen groups (primary and metastatic lesions) were analysed. -Fetoprotein (AFP) was also determined in transplanted tumours by an immunohistochemical method. Orthotopic growth was observed in 14 of 30 transplanted specimens and formation of metastases in 7 cases, which exhibited the variety of clinical behaviours seen in patients with HCC. These behaviours included local growth, regional invasion, spontaneous intrahepatic, lymph node and lung metastasis and peritoneal seeding. In two groups the growth rate of metastatic lesions following implantation was clearly higher than that of primary tumours. Chromosome analysis from locally growing tumours confirmed their morphologically human origin. An immunohistochemical study showed that implanted tumours originating from AFP-positive specimens maintained AFP expression. These results indicated that the animal models should prove valuable for developing new treatment modalities and studying the mechanism of metastasis of human HCC.Abbreviation HCC hepatocellular carcinoma - AFP -fetoprotein     

Inhibitory effect of the angiogenesis inhibitor TNP-470 on tumor growth and metastasis in nude mice bearing human hepatocellular carcinoma

The antitumor and anti-metastatic effects of a potent angiogenesis inhibitor,O-(chloroacetyl-carbamoyl)fumagillol (TNP-470), was investigated in a highly metastatic model of human hepatocellular carcinoma—LCI-D20. Small pieces of LCI-D20 tumor tissue were implanted subcutaneously into the right axillary region of 24 nude mice; the mice were then randomized into two groups. To one group, TNP-470 30 mg/kg was given as a subcutaneous injection every other day from day 1 to day 15 and the mice were sacrificed on day 26. An antitumor effect of TNP-470 was clearly demonstrated by tumor weight (0.97±0.34 g compared to 2.04±0.34 g,P<0.001) and -Fetoprotein value (93±59 g/L compared to 769±282 g/L,P<0.001). There was also an anti-metastatic effect of TNP-470. Lung metastases developed in only 1 of 12 mice in the treated group, while they developed in 6 of mice of the control group. No severe side-effect of TNP-470 was found in this study. In vitro study revealed that the purified hepatoma cells were insensitive to TNP-470 (the 50% inhibitory concentration was 43 g/ml). These results suggest that the angiogenesis inhibitor TNP-470 has both strong antitumor and anti-metastatic effects on a human hepatocellular carcinoma model in nude mice.Abbreviations TNP-470 O-(chloroacetyl-carbamoyl) Fumagillol - MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide This work was partly supported by the CHina Medical Board of New York, grant 93-583, and a Leading Speciality grant of Shanghai Health Bureau     

Effects and mechanisms of silibinin on human hepatocellular carcinoma xenografts in nude mice

AIM： To investigate the in vivo effects and mechanisms of silibinin on the growth of hepatocellular carcinoma （HCC） xenografts in nude mice. METHODS： Nude mice bearing HuH7 xenografts were used to assess the anti-HCC effects and mechanisms of silibinin. RESULTS： Silibinin resulted in a potent dosedependent reduction of HuH7 xenografts in association with a significant decrease in Ki-67 and α-fetoprotein production, nuclear NF-κB content, polo-like kinase 1, Rb phosphorylation, and E2F1/DP1 complex, but increased p27/CDK4 complex and checkpoint kinase 1 expression, suggesting that the in vivo effects of silibinin are mediated by inhibiting G1-S transition of the cell cycle. Silibinin-induced apoptosis of HuH7 xenografts was associated with inhibited survivin phosphorylation. Silibinin-reduced growth of HuH7 xenografts was associated with decreased p-ERK, increased PTEN expression and the activity of silibinin was correlated with decreased p-Akt production, indicating involvement of PTEN/PI3K/Akt and ERK pathways in its in vivo anti-HCC effects. Silibinin-reduced growth of HuH7 xenografts was also associated with a significant increase in AC-H3 and AC-H4 expression and the production of superoxide dismutase （SOD）-1.CONCLUSION： Silibinin reduces HCC xenograft growth through the inhibition of cell proliferation, cell cycle progression and PTEN/P-Akt and ERK signaling, inducing cell apoptosis, and increasing histone acetylation and SOD-1 expression.     

Development and characterization of multidrug resistant human hepatocarcinoma cell line in nude mice

INTRODUCTION Tumor cell drug resistance represents a signif icant obstacle to successful chemotherapy. Cells which have acquired resistance to one anti-tumor drug usually show resistance to other anti-tumor drugs[1]. This cellular resistance is known as m…     

Serum CYFRA 21-1 level reflects hepatocellular carcinoma metastasis: study in nude mice model and clinical patients

Our previous proteomics study on human hepatocellular carcinoma (HCC) cell strains revealed that cytokeratin 19 (CK19) was expressed in cells with high metastasis potential; we further studied serum CK19 fragment CYFRA 21-1 level in HCC patients and nude mice model of HCC metastasis. HCC cell line HCCLM3 was injected subcutaneously into 30 nude mice which were then randomized into 6 groups of 5 mice each. The murine serum CYFRA 21-1 and pulmonary metastases were determined 2, 3, 4, 5, 6, and 7 weeks after injection. Serum CYFRA 21-1 levels of 101 normal controls and 108 HCC patients were also determined. In nude mice model, CYFRA 21-1 level increased significantly when pulmonary metastases occurred. Among 108 HCC patients, 24 (22.2%) had increased serum CYFRA 21-1 level. The presence of portal vein tumor emboli was significantly higher in CYFRA 21-1 increased cases (33.3%, 6/24) than in CYFRA 21-1 normal cases (6.0%, 5/84) (x ² = 7.403, P < 0.01). In addition, the percentage of TNM stage III/IV tumor was significantly higher in CYFRA 21-1 increased patients (54.2%, 13/24) than in CYFRA 21-1 normal cases (21.4%, 18/84) (x ² = 9.776, P < 0.005). These results suggest that CK19 may play an important role in HCC metastasis.     

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.     

M2型丙酮酸激酶在肝细胞癌中的研究进展

原发性肝细胞癌是世界上常见、恶性程度很高的肿瘤之一,在中国恶性肿瘤中发病率排第5位,肝癌的各种治疗措施疗效不佳。M2型丙酮酸激酶作为糖酵解途径关键酶,它的异常表达与肝癌的增殖、转移、诊断、治疗及预后密切相关,与肝癌药物治疗的耐药和放射线抗拒也有关联,采用多种途径靶向调控肝癌细胞的M2型丙酮酸激酶,有望成为治疗原发性肝癌...