GHGKHKNK八肽抑制小鼠恶性黑色素瘤B16-F10转移及其机制的实验研究

目的：探讨GHGKHKNK八肽抑制小鼠恶性黑色素瘤B16-FIO细胞转移及其机制。结果：1．不同剂量的GHGKHKNK八肽对小鼠恶性黑色素瘤细胞B16-FIO的克隆形成均有抑制作用,与对照组相比较均有显著性差异（P〈0．05）2．不同剂量的GHGKHKNK八肽在体外作用24h后可以显著抑制小鼠恶性黑色素瘤B16-FIO细胞的侵袭,穿膜细胞数与对照组比较具有显著性差异（P〈0．05）。3．GHGKHKNK八肽在24h时5．5×10^-4mol／L、5．5×10^-5mol／L、5．5×10^-6mol／L对小鼠恶性黑色素瘤细胞B16-F10黏附抑制率与对照组相比较均有显著性差异（P〈0．05）。4．经GHGKHKNK八肽作用后,小鼠恶性黑色素瘤细胞ICAM-1、LN—R表达降低表达降低,E-cadhesion表达增强。与对照组比较明显差异P〈0．05．5．GHGKHKNK八肽对B16一F10人工肺转移的抑制作用的实验结果显示：GHGKHKNK八肽各剂量组和环磷酰胺组肺转移灶的平均数目与生理盐水组照组相比较有明显差异（P〈0．05）：GHGKHKNK八肽500ug／kg／d、50ug／kg／d剂量组与环磷酰胺组对比有显著性差异（P〈0．05）。结论：i．GHGKHKNK八肽在体外能抑制小鼠恶性黑色素瘤细胞的体外克隆形成能力,能显著抑制小鼠恶性黑色素瘤B16-F10细胞的侵袭。提示HGKHKNK八肽可能通过抑制小鼠恶性黑色素瘤细胞的侵袭和增殖而抗肿瘤细胞的转移。2．GHGKHKNK八肽抑制小鼠小鼠恶性黑色素瘤细胞B16-FIO与人工基底膜的黏附,提示GFIGKHKNK八肽可能通过抑制细胞与基底膜的黏附来抑制肿瘤细胞的转移。3．GHGKHKNK八肽能够下调细胞间黏附因子1（ICAM-1）和层黏连蛋白受体（LN—R）在细胞内的表达,提示GHGKHKNK八肽可能通过抑制细胞间黏附因子的表达而抑制肿瘤细胞的转移。4．GHGKHKNK八肽能显著抑制小鼠恶性黑色素瘤细胞B16-F10在小鼠体内的肺转移。     

氨氯地平抑制小鼠黑色素瘤高转移细胞株B16体内转移的作用及相关机制研究

目的:研究不同剂量氨氯地平抑制小鼠黑色素瘤高转移细胞株B16转移的作用,并探讨其作用机制。方法:选取64只C57BL/6J小鼠,将黑色素瘤髙转移细胞株B16接种在小鼠腹股沟皮下(2×106个/只)。将接种后的小鼠以随机数字表法分为对照组和氨氯地平高、中、低剂量治疗组,对照组小鼠仅给予0.9%氯化钠注射液;治疗组小鼠给予不同剂量的氨氯地平治疗,分别为1 mg/kg(低剂量组)、3 mg/kg(中剂量组)和10 mg/kg(高剂量组)。观察各组小鼠肿瘤生长情况及身体情况;观察各组小鼠肿瘤肺转移情况,计算肺转移的结节及肿瘤抑瘤率;采用免疫组织化学法检测各组小鼠肿瘤组织中白细胞介素8(IL-8)蛋白表达水平;采用逆转录-聚合酶链反应(RT-PCR)检测各组小鼠肿瘤组织中IL-8的信使核糖核酸(mRNA)表达水平。结果:氨氯地平对小鼠黑色素瘤高转移细胞株B16的自发性肺转移有显著的抑制作用;治疗组小鼠肿瘤组织中IL-8蛋白表达均明显减弱,且随氨氯地平剂量的增加而减弱;治疗组小鼠肿瘤组织中IL-8的mRNA的表达明显低于对照组,且IL-8的mRNA的表达随氨氯地平剂量的增加而减弱。结论:氨氯地平对小鼠黑色素瘤高转移细胞株B16有一定的抑瘤和抑制肺转移作用,其作用可能与影响IL-8的表达有关。     

国产云芝多糖对小鼠抗肿瘤免疫反应的促进作用

本文采用观察同一个体多种指标的方法,研究国产云芝胞内多糖对C57BL/6J/J小鼠体内的黑色素瘤B16的抗瘤作用及其促进荷瘤鼠抗肿瘤免疫反应的作用.结果表明,这种云芝多糖对黑色素瘤B16的抑瘤率为40.8%,与环磷酰胺合用对黑色素瘤B16人工肺转移的抑制率达84.4%;其抗瘤作用机理以拮抗肿瘤的免疫抑制作用,多方面有效地促进行瘤鼠的非特异性抗肿瘤免疫反应为主,对正常机体的免疫反应也有一定促进作用.     

脂质体干扰素α对小鼠Lewis肺癌转移及脾细胞增殖的影响

C57BL／6N小鼠尾静脉注射Lewis肺癌细胞后，腹腔注射脂质体干扰素α（L-IFN－α）和未包裹干扰素α（IF－α），连续10d；测小鼠肺湿重、肺转移给节数和ConA诱导的脾细胞DNA参入量。结果IFN－α5×104、20×104U／kg组，小鼠肺湿重较对照组减少14．1％、19．6％，肿瘤肺转移结节数减少32．7％、50.0％，ConA刺激的脾细胞DNA参入量增加24．6％、46．8％。1／10剂量的L-IFN－α可产生相同的药理活性。小鼠环磷酰胺50mg／kgip3d后，尾静脉注射瘤细胞，实验处理同上。结果L-IFN-α和IFN－α对环磷酰胺处理小鼠的Lewis肺癌细胞肺转移的抑制作用更为明显。本文结果表明，IFN－α可抑制小鼠Lewis肺癌肺转移，促进ConA刺激的脾细胞增殖，并与剂量明显相关。脂质体包裹IFN－α可使其生物活性提高约10倍。对免疫功能受抑小鼠作用更为显著。     

鲨骨精抗黑色素瘤肺转移及调节免疫功能的研究

鲨骨精(shark cartilage active extract．SCAE)是鲨鱼软骨提取物。本项研究评价其抗黑色素瘤B16肺脏转移及调节机体免疫功能的作用。小鼠口服SACE后d7，经尾静脉注射黑色素细胞瘤B16细胞悬液；3w后计数黑色素瘤肺脏转移结节，测定脾细胞的免症功能。结果表明，SCAE能抑制黑色素瘤B16肺转移，抑制率为33%能提高脾脏有核细胞总数、淋巴细胞增殖能力和LAK细胞活性。证实了SCAE能抑制黑色素癌转移及提高机体免疫功能。     

三七总皂苷对B16黑色素瘤生长及转移的作用

目的观察三七总皂苷（PNS）对B16黑色素瘤生长及转移的作用,及其对B16黑色素瘤表达缝隙连接蛋白Connexin32的影响。方法实验采用B16黑色素瘤自发性肺转移模型,来观察PNS对B16黑色素瘤生长及转移的影响。免疫组织化学检测Connexin32在黑色素瘤原发灶的表达情况。结果（1）PNS对黑色素瘤生长有较好的抑制作用,其中PNS高剂量组抑瘤率可达50．85％。（2）与模型组相比PNS中、高剂量组能有效抑制黑色素瘤的肺转移,转移灶数量与对照组相比有明显减少。（3）免疫组织化学检测,发现PNS各用药组均能增强黑色素瘤原发灶细胞膜上Connexin32的表达。结论PNS能够抑制B16黑色素瘤的生长和转移,并能有效增强肿瘤细胞膜上Connexin32的表达。     

rhIL-24对黑色素瘤B16细胞的体内外抑瘤作用

目的 研究重组人白细胞介素24（rhIL-24）及真核表达基因重组质粒pcDNA3．0-hIL-24对黑色素瘤B16细胞（B16细胞）及其荷瘤小鼠体内外抑肿瘤作用。方法 用rhIL-24蛋白作用于小鼠B16细胞,检测其对B16细胞的生长抑制作用及诱导凋亡作用,并用rhIL-24蛋白及pcDNA3．0-hIL-24治疗荷瘤小鼠。结果 rhIL-24蛋白对体外培养的B16细胞具有显著的抑制生长和诱导凋亡作用,rhIL-24蛋白及pcDNA3．0-hIL-24对C57／BL6小鼠体内形成的恶性黑色素瘤具有明显的抑瘤作用。结论 rhIL-24蛋白具有生物活性,他及真核表达重组质粒对B16细胞及其小鼠种植瘤生长有抑制作用,并能诱导肿瘤细胞凋亡及促进其分化。     

土贝母苷甲对小鼠B16黑色素瘤和Lewis肺癌转移的抑制作用

目的:本研究旨在探讨土贝母苷甲对小鼠B16黑色素瘤和Lewis肺癌转移的影响。方法:采用裸BALB/c小鼠B16黑色素瘤实验转移和裸BALB/c小鼠Lewis肺癌自发转移模型研究土贝母苷甲对肿瘤转移的影响;采用免疫组织化学法检测土贝母苷甲对转移相关基因CD44v6、ErbB-2和nm23-H1表达的影响。结果:腹腔注射低于全身中毒水平剂量的土贝母苷甲明显减少接种B16黑色素瘤细胞小鼠的肺重和肺转移灶数量,而对小鼠的健康和活力无明显影响;土贝母苷甲(2、3 mg.kg-1.d-1×14 d)和环磷酰胺(50 mg.kg-1.wk-1×2 d)对B16黑色素瘤细胞肺转移的抑制率分别为68.8%、82.8%和49.1%。土贝母苷甲治疗组小鼠Lewis肺癌原发肿瘤的平均瘤重明显低于对照组(P<0.05)。土贝母苷甲(2、3 mg.kg-1.d-1×14 d)和环磷酰胺(50 mg.kg-1.w-1×2 d)治疗组小鼠的肝转移抑制率分别为46.3%、52.0%和54.7%。土贝母苷甲显著下调促转移基因CD44v6和ErbB-2的表达,上调抑转移基因nm23-H1的表达。结论:土贝母苷甲对小鼠B16黑色素瘤的实验性转移和Lewis肺癌的自发性转移都有显著的抑制作用。土贝母苷甲对肿瘤转移的这种抑制作用与其抑制原发肿瘤生长、下调促转移基因CD44v6和ErbB-2及上调抑转移基因nm23-H1的表达有关,提示土贝母苷甲是一个阻止癌扩散和转移的候补化合物。     

弓形虫溶解抗原抗小鼠B16黑色素瘤血管生成的实验研究

目的 观察弓形虫溶解抗原(TLA)对小鼠B16黑色素瘤生长以及肿瘤血管生成的影响.方法 20只C57BL/6J小鼠皮下接种B16黑色素瘤细胞,建立荷瘤动物模型,从小鼠荷瘤的第7天开始,每隔2天实验组小鼠经腹腔注射TLA 0.1 ml,对照组小鼠注射同等剂量0.9%氯化钠注射溶液,荷瘤21 d后处死小鼠剥取肿瘤,测量肿瘤体积与重量,计算抑瘤率,免疫组化法检测肿瘤微血管密度(MVD)与血管内皮生长因子(VEGF)表达情况.结果 TLA显著抑制了小鼠体内黑色素瘤生长,实验组肿瘤体积与重量明显小于对照组(P＜0.05),抑瘤率为49.6%.实验组与对照组的MVD值分别为(44.4000±4.7888)、(31.9000±2.6012),两组相比较差异有统计学意义(P＜0.05).实验组肿瘤VEGF表达明显低于对照组(P＜0.05).结论 TLA能够抑制小鼠B16黑色素瘤生长,其抗瘤机制可能与抗血管生成有关.     

甘草酸二铵磷脂复合物对小鼠慢性肝损伤的保护作用

目的：探讨甘草酸二铵磷脂复合物注射剂（DG-PC）对C57BL/6J小鼠慢性免疫性肝脏损害的保护作用。方法：复制C57BL/6J小鼠慢性免疫性肝脏损害模型,测定血清ALT、AST、蛋白和透明质酸（HA）,肝匀浆中超氧化物岐化酶（SOD）、谷胱甘肽（GSH）、I型胶原（C.I）,并对肝脏组织切片进行电镜观察。结果：DG-PC可以明显改善肝损小鼠AST水平、提高白蛋白（ALB）含量、升高白球比（A/G）、降低HA含量（P〈0.01）,提高肝损小鼠的SOD和GSH水平（P〈0.01）,治疗效果优于甘草酸二铵（DG）。小鼠肝组织透射电镜观察显示DG-PC可以保护肝细胞和细胞器,减轻纤维化程度。结论：DG-PC对于C57BL/6J小鼠慢性免疫性肝损伤均具有一定的保护作用,可能与抗氧化等活性有关。     

胡桃醌对小鼠黑色素瘤细胞B16F10体内迁移的影响

目的:研究胡桃醌对小鼠黑色素瘤细胞B16F10体内迁移的影响.方法:采用小鼠B16/F10黑色素瘤人工肺转移模型研究胡桃醌对肿瘤细胞血道转移的作用.结果:与溶剂对照组相比,4.5、3、1.5和0.75 mg/kg胡桃醌组显著减少小鼠黑色素瘤B16F10血道转移(P＜0.05),其抑制率分别为27.30％、55.35％、31.52％和25.34％;3 mg/kg胡桃醌与阳性对照组相比,抑瘤率无统计学差异(P＞0.05).结论:胡桃醌可抑制小鼠黑色素瘤B16F10血道转移.     

Antimetastatic activity of acteoside,a phenylethanoid glycoside

We examined the antimetastatic effect of acteoside, a phenylethanoid glycoside widely distributed in the plant kingdom, on lung metastasis using a mouse model injected with B16 melanoma cells intravenously. Male C57BL/6 mice were injected intravenously with 2 x 10(5) of B16 melanoma cells, while acteoside at a dose of 50 mg/kg was administered intraperitoneally every other day from 13 d before B16 melanoma cell injection until all mice had succumbed to the metastatic tumor burden in the lung. Administration of acteoside prolonged survival time significantly and the average survival time was 63.3 +/- 3.4d compared with 52.1 +/- 2.5d in control mice. This result suggests that acteoside showed suppressive effect on lung metastasis of B16 melanoma cells.     

Ginkgo biloba exocarp extracts inhibits metastasis and its effects on TGF-β1/ERK1/2/MMP-9 signaling pathway in B16-F10 melanoma

OBJECTIVE To study the inhibition and mechanism of Ginkgo biloba exocarp extracts(GBEE)for the metastasis of B16-F10 melanoma in C57 BL/6 J mice. METHODS The metastasis model of B16-F10 in C57BL/6J mice was set up. The C57 BL/6 J mice were randomly separated into these groups: positive control,model control, normal control and GBEE treatment groups, n=10. The mice in positive group wereintraperitoneal(ip) injectioncis-dichlorodiamineplatinum(Ⅱ) at a dose of 5 mg·kg~(-1), twice a day for 7 d; model group and normal group were both intragastric gavage(ig) normal saline(NS) in a volume of 0.1 m L/10 g, once a day for17 d; the GBEE treatment groups were respectively ig GBEE 50, 100 and 200 mg · kg~(-1), once a day for 17 d.After the administration, the lung tissue was removed and the lung surface metastasis was observed; the rate of lung metastasis and anti-metastasis were calculated;the degree of lung metastasis was observed by HE staining; in vitro, the effect of GBEE on the migration rate of B16-F10 cel s was detected by wound healing test; Western Blot was used to detect the expression of TGF-β_1, ERK1/2,p-ERK1/2 and MMP-9 protein in B16-F10 cel s. RESULTS In vivo, we discovered that GBEE(50, 100, 200 mg·kg~(-1))cansuppress tumor lung metastasis of B16-F10 melanoma in C57 BL/6 J mice in a dose-dependent way. In vitro, we found that GBEE(20, 40, 80 mg·L~(-1)) can significantly inhibit B16-F10 cells treated for 24 h and 48 h migration in a time-and concentration-dependent way. GBEE(20, 40,80 mg ·L~(-1)) can suppressed TGF-β_1, p-ERK1/2/ERK1/2 and MMP-9 protein expression level in a concentrationdependent way. CONCLUSION GBEE significantly inhibit the metastasis of B16-F10 melanoma, and its mechanism of action is related to the inhibition of extracellular matrix degradation and cell migration through the TGF-β_1/ERK1/2/MMP-9 signaling pathway.     

Possible reduction of hepatoma formation by Smmu 7721 cells in SCID mice and metastasis formation by B16F10 melanoma cells in C57BL/6 mice by Agaricus blazei murill extract

Agaricus blazei Murill extract (ABM) has been reported to possess antitumor effects. In this study, the role of ABM in tumor growth and metastasis in vivo was evaluated in experimental Smmu 7721 hepatoma cells in severe combined immunodeficiency (SCID) mice and B16F10 melanoma cells lung metastasis in C57BL/6 mice. For the tumor growth model, the size of the liver tumor mass was about 10 mm to 20 mm in the control group. In comparison with the control group, the tumor mass seem to grow slowly with ABM treatment, especially at the high dose. For the tumor metastasis model, after a six-week treatment, the survival rates of B6 mice were 0%, 30%, 10% and 50% for control group, low, median and high concentration ABM treatment groups, respectively. The survival rate showed that pretreatment of C57BL/6 (B6) mice with ABM lengthened their lifespan after tumor cell inoculation, which supports the notion that ABM successfully reduced lung metastasis formation by B16F10 melanoma cells. The treatment effect was dependent on the concentration of ABM for tumor growth and metastasis in these models.     

重组人纤连蛋白CH50对黑色素瘤生长及转移的影响

目的：研究重组人纤连蛋白ＣＨ５０对小鼠黑色素瘤生长的影响及其抗肿瘤机制。方法 进行小鼠动物实验及黑色素瘤Ｂ１６细胞实验。结果：ＣＨ５０在体内明显抑制黑色素瘤生长及实验性肺转移,ＣＨ５０在体外能附粘黑色素瘤Ｂ１６细胞,抑制Ｂ１６细胞粘附层粘蛋白并明显提高腹腔巨噬细胞对Ｂ１６细胞杀伤活性,结论：ＣＨ５０抑制小鼠黑色素瘤的生长和转移,ＣＨ５０的抗肿瘤机制与其粘附肿瘤细胞,提高巨噬细胞杀瘤性活性有关。     

Antimetastatic potential of vernolide-A, a sesquiterpenoid from Vernonia cinerea L

The inhibitory effect of vernolide-A (C(21)H(28)O(7)) on lung metastasis induced by B16F-10 melanoma cells was studied using C57BL/6 mice. Vernolide-A was administered in three different modalities such as simultaneously with tumor, prophylactic to tumor and after tumor development. Maximum inhibition in the metastasis was observed when vernolide-A was administered simultaneously with tumor. There was 89.39% inhibition of lung tumor nodule formation and 88.51% increase in the life span of metastatic tumor-bearing animals. Highly elevated levels of lung hydroxyproline, lung uronic acid, lung hexosamine, serum sialic acid, serum γ-glutamyl transpeptidase (GGT) and serum vascular endothelial growth factor (VEGF) in the metastatic control animals were found to be significantly lowered in the vernolide-A-treated animals. Histopathological analysis of lung tissues also correlated with these results. Vernolide-A administration downregulated the expression of matrix metalloproteinase-2 (MMP-2), MMP-9, extracellular-signal-regulated kinase-1 (ERK-1), ERK-2 and VEGF in the lung tissue of B16F-10 melanoma challenged animals. In the in vitro system, vernolide-A showed a significant inhibition of invasion of B16F-10 melanoma cells across the collagen matrix. Vernolide-A treatment also inhibited the migration of B16F-10 melanoma cells across a polycarbonate filter in vitro. Vernolide-A could inhibit MMP-2 and MMP-9 protein expression in gelatin zymographic analysis of B16F-10 cells. (3)H-thymidine proliferation assay showed that vernolide-A could inhibit the proliferation of B16F-10 melanoma cells in vitro. These results indicate that vernolide-A could inhibit the metastatic progression of B16F-10 melanoma cells in mice.     

海参岩藻聚糖硫酸酯抗肿瘤转移作用研究

目的建立小鼠黑色素瘤B16细胞实验性肺转移模型,探讨海参岩藻聚糖硫酸酯(sea cucumber fu-coidan,SC-FUC)的体内抑制肿瘤肺转移作用。方法连续腹腔注射SC-FUC 26d后,检测小鼠肺转移灶数量、血清中唾液酸的含量、γ-谷氨酰转肽酶活力和肺组织中羟脯氨酸、氨基己糖、糖醛酸的含量。结果SC-FUC剂量组小鼠的肺转移灶数量显著减少(P<0.01),平均转移抑制率为65.25%,血清唾液酸含量和γ-谷氨酰转肽酶活力显著降低(P<0.01),肺组织中羟脯氨酸、氨基己糖、糖醛酸的含量显著下降(P<0.01)。结论SC-FUC能显著抑制肿瘤细胞在小鼠体内的转移和生长。     

Inhibitory effect of BCG cell-wall skeletons (BCG-CWS) emulsified in squalane on tumor growth and metastasis in mice

The antimetastatic effect of BCG-CWS, which was emulsified in an oil-in-water form with either Drakeol 6VR mineral oil (BCG-CWS/DK) or squalane (BCG-CWS/SQA), on lung metastasis produced by highly metastatic murine tumor cells, Colon26-M3.1 carcinoma cells and B16-BL6 melanoma cells, was investigated in syngeneic mice. An intravenous (i.v.) administration of BCG-CWS (100 mg/mouse) 1 day after tumor inoculation significantly inhibited tumor metastasis of both Colon26-M3.1 carcinoma and B16-BL6 melanoma cells in experimental lung metastasis models. No differences in the antitumor activity of the two oil-based formulations (BCG-CWS/DK and BCG-CWS/SQA) were obverved. However, BCG-CWS/SQA administered through subcutaneous (s.c.) route was shown to be effective only when it was consecutively injected (3 times) after tumor inoculation. An in vivo analysis for tumor-induced angiogenesis showed that a single i.v. administration of BCG-CWS/SQA inhibited the number of tumor-induced blood vessels and suppressed tumor growth. Furthermore, the multiple administration of BCG-CWS/SQA given at on week intervals led to a significant reduction in spontaneous lung metastasis of B16-BL6 melanoma cells in a spontaneous metastasis model. These results suggest that BCG-CWS emulsified with squalane is a potent inhibitory agent of lung metastasis, and that the antimetastatic effect of BCG-CWS is related to the suppression of tumor growth and the inhibition of tumor-induced angiogenesis.