鱼腥草黄酮提取物对肿瘤细胞的抑制作用

目的研究鱼腥草黄酮类提取物在体外对人白血病细胞HL60、小鼠黑色素瘤细胞株B16BL6增殖抑制和凋亡的影响.方法(1)采用纯化水回流提取,大孔树脂分离的方法提取鱼腥草中黄酮类化合物;(2)应用MTT法检测黄酮类化合物对HL60和B16BL6肿瘤细胞株的细胞抑制率;(3)采用FCM法检测其对肿瘤细胞凋亡的影响.结果(1)鱼腥草黄酮提取物有抑制HL60和B16BL6细胞生长的作用,50%增值抑制浓度值分别为0.410,0.122 g·L-1,(2)鱼腥草黄酮提取物能诱导HL60和B16BL6细胞的凋亡.结论鱼腥草黄酮提取物能抑制HL60和B16BL6肿瘤细胞生长,具诱导凋亡的作用,为鱼腥草的抗肿瘤效应提供实验依据.     

女贞子多糖对黑色素瘤细胞黏附能力的影响

目的探讨女贞子多糖对黑色素瘤细胞B16BL6黏附能力的影响。方法观察女贞子多糖对黑色素瘤细胞黏附力的抑制作用,分别将琼脂糖和Matrigel铺于细胞板中,用女贞子多糖作用后,MTT法观察女贞子多糖对B16BL6黏附能力的影响;同时观察了女贞子多糖对黏附分子E-cadherin表达的影响。结果女贞子多糖抑制了黑色素瘤细胞的粘附能力,抑制黑色素瘤粘附分子E-cadherin的表达。结论女贞子多糖具有抑制黑色素肿瘤细胞的黏附,破坏了肿瘤细胞的整体性,利于女贞子多糖的抗肿瘤作用。     

尖尾芋醇提物保护黑色素瘤小鼠脾的作用机制

目的 研究尖尾芋醇提物对荷瘤小鼠脾保护的作用机制.方法 C57BL/6小鼠背部右侧皮下接种黑色素瘤B16-F10细胞后,将未造模的6只小鼠作为正常组(normal),造模的30只小鼠随机分为模型组(model)、环磷酰胺组(CTX)和低、中、高剂量的尖尾芋醇提物组(lEAC、mEAC、hEAC),每组各6只.连续给药1...     

水解贝壳硬蛋白美白功效的初步研究

研究水解贝壳硬蛋白(Shell protein extract,SPE)对B16细胞内黑色素含量及酪氨酸酶活性的影响.培养B16细胞,构建α-黑素细胞刺激素(α-MSH)诱导细胞的黑色素高表达量细胞模型.SPE作用B16细胞,CCK8法测定细胞活力;NaOH裂解法和L-DOPA氧化法测定细胞内黑色素含量和酪氨酸酶活性的...     

Lewis肺癌小鼠Th1/Th2免疫应答状态的观察

目的研究Lewis肺癌小鼠Th1/Th2免疫应答状态及其意义。方法C57BL/6小鼠16只按随机数字法分为肺癌模型组(A组)与正常对照组(B组),每组8只。A组采用Lewis肺癌单细胞悬液接种于C57BL/6小鼠右腋部皮下建立肺癌模型;B组则正常饲养未予处理。两组实验动物于实验末取血,应用酶联免疫吸附法(ELISA)检测血清中Th1细胞因子IFN-γ、Th2细胞因子IL-4浓度,并进行组间相关比较。结果A组小鼠血清中Th1相关细胞因子IFN-浓度明显低于B组小鼠,A组小鼠Th2相关细胞因子IL-4浓度明显高于B组小鼠,差异均有统计学意义(P<0.05);而FN-γ/IL-4比值A组小鼠相应低于B组小鼠,差异亦有统计学意义(P<0.05)。结论Lewis肺癌小鼠Th1/Th2免疫应答状态呈现明显Th1向Th2漂移,使肺癌细胞逃避机体免疫监视,并削弱机体的细胞免疫功能,这应是肿瘤发生发展的主要原因之一。     

rhddADAM15蛋白抑制小鼠黑色素瘤细胞生长及其机制

目的探讨重组人ADAM15去整合素结构域蛋白(rhddADAM15)对小鼠黑色素瘤细胞B16体外增殖、迁移、侵袭和凋亡等细胞行为的影响以及对p38丝裂原激活蛋白激酶(p38MAPK)信号通路的作用。方法 MTT法检测rhd-dADAM15对B16细胞增殖的抑制作用;划痕实验观察rhdd-ADAM15对B16细胞迁移的影响;"侵袭小室法"检测rhdd-ADAM15对B16细胞侵袭的作用;流式细胞术分析细胞周期变化;Western blot法检测rhddADAM15作用导致的p38MAPK激酶活性的变化。结果 rhddADAM15作用明显抑制B16细胞生长(IC50为13.80 mg.L-1),当浓度为7.5mg.L-1时,侵袭细胞数较对照组减少了0.55,对B16细胞迁移的抑制作用与共培养的时间呈正比,主要将B16细胞生长阻滞在G2/M期;当rhddADAM15浓度为10 mg.L-1时,p38MAPK激酶的磷酸化程度达0.80。结论 rhddADAM15对B16细胞行为具有明显的抑制作用,其机制可能与激活p38MAPK信号通路有关。     

小鼠皮下移植瘤冷冻消融实验中用于形成移植瘤的种子细胞数量的选择

目的 观察接种不同细胞数量下C57BL/6小鼠Lewis肺癌(LLC)皮下移植瘤瘤体变化,选择适合冷冻消融实验的种子细胞数量.方法 将18只6～8周龄的C57BL/6小鼠随机分为三组,每组6只,在右上肢腋窝皮下接种LLC细胞,数量分别为1×106个(A组)、2×106个(B组)和5×106个(C组).比较三组移植瘤成瘤...     

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.     

吡唑啉酮衍生物镉(Ⅱ)配合物体内外对黑素瘤B16细胞的抗肿瘤作用

目的研究吡唑啉酮镉(Ⅱ)配合物1-苯基-3-甲基-4-丙酰基-5-吡唑啉酮缩水杨酰肼-镉(Ⅱ)(Cd-PMPP-SAL)体内外对小鼠黑素瘤B16细胞的抗肿瘤作用及其作用机制。方法以Cd-PMPP-SAL1.0,1.5,3.0,5.0和10.0 mg·L~(-1)分别作用小鼠黑素瘤B16细胞24,48和72 h,采用MTT法检测B16细胞存活率;Cd-PMPP-SAL 6.25,12.50和25.00 mg·L~(-1)作用B16细胞24 h,用Hoechst33258染色观察B16细胞形态,AnnenxinⅤ/PI双染色法检测B16细胞凋亡率;胱天蛋白酶活性检测试剂盒检测B16细胞内胱天蛋白酶活性。C57BL/6J小鼠皮下接种B16细胞制备荷瘤模型,5 d后分别瘤内注射Cd-PMPP-SAL 6.25,12.50和25.00 mg·kg~(-1),每天1次,连续12 d。每天检测体质量,给药结束后处死小鼠,测量瘤体积并测瘤质量,计算抑瘤率。HE染色法观察瘤体、肝和肺组织病理变化;免疫组织化学法检测肿瘤组织中血管内皮生长因子(VEGF)和成纤维细胞生长因子2(FGF2)蛋白表达;TUNEL法检测移植瘤组织内的细胞凋亡。结果Cd-PMPP-SAL抑制B16细胞存活,IC_(50)为4.946 mg·L~(-1),95%置信限为4.24~5.65 mg·L~(-1);Cd-PMPP-SAL12.50和25.00 mg·L~(-1)作用24 h,B16细胞凋亡率为(12.8±1.4)%和(18.4±0.4)%,显著高于细胞对照组(1.7±0.1)(P<0.01);Cd-PMPP-SAL 25.00 mg·L~(-1)组胱天蛋白酶3和9活性与细胞对照组比较显著增高(P<0.01),胱天蛋白酶3/7活性变化不明显。瘤内注射Cd-PMPP-SAL 12.50和25.00 mg·kg~(-1)治疗组,从治疗第8天起瘤体积与模型组相比明显减小(P<0.01),对小鼠体质量无明显影响;Cd-PMPP-SAL 12.50和25.00 mg·kg~(-1)治疗组小鼠移植瘤组织有不同程度的坏死,肝、肺组织无明显病理变化;与模型组比较,Cd-PMPP-SAL 12.50和25.00 mg·kg~(-1)治疗组移植瘤组织VEGF和FGF2蛋白表达显著下降(P<0.05),凋亡细胞明显增加(P<0.05)。结论 Cd-PMPP-SAL体内外可有效地抑制B16细胞生长,该作用可能与诱导细胞凋亡及抑制肿瘤内血管生成有关。     

三氧化二砷抑制小鼠B16黑色素瘤生长作用及其机制

目的　探讨三氧化二砷 (As2 O3 )对小鼠B16黑色素瘤的生长及其血管生成的抑制作用 ;同时观察其对B16细胞增殖活性、细胞形态、细胞周期及凋亡的影响。方法　选用小鼠黑色素瘤B16细胞接种C5 7BL/ 6J小鼠 ,观察腹腔注射As2 O3 对实体瘤的重量及成瘤率的影响 ;应用HE染色、Ⅷ RAg免疫组化染色观测瘤组织内新生血管密度 ;采用CellTiter 96AqueousOne试剂检测B16细胞增殖活力 ;Giem sa染色、Feulgen染色观察细胞形态学变化 ;流式细胞术分析细胞周期及细胞凋亡。结果　As2 O3 能显著抑制小鼠B16黑色素瘤的生长 ,治疗组成瘤率为 37 5 % ,抑瘤率达81 6 1% ,并能显著抑制瘤组织内血管生成 ;体外实验观察到As2 O3 能抑制B16细胞增殖 ,并存在浓度依赖效应 ,IC50 为32 99μmol·L-1;细胞形态学观察结果显示As2 O3 使B16细收稿日期 :2 0 0 4-0 2 -17,修回日期 :2 0 0 4-0 3 -2 8基金项目 :安徽省教育厅资助项目 ,No 2 0 0 4kJ2 79作者简介 :夏　俊 ( 1965 -) ,女 ,硕士 ,副教授 ,硕士生导师 ,研究方向 :肿瘤分子生物学 ,Tel:0 5 5 2 3 0 664 12 2 0 97,E mail:xia jun1965 @yahoo .com .cn崔秀云 ( 1941-) ,女 ,教授 ,博士生导师 ,研究方向 :癌基因与抑癌基因 ,Tel:0 411 472 0 64 8,E mail:cuixy @dlmedu .edu .     

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.     

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.     

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.     

金荞麦提取物抑制肿瘤细胞侵袭、转移和HT-1080细胞产生Ⅳ型胶原酶的研究

目的探讨金荞麦提取物对肿瘤细胞侵袭和转移的影响。方法以人工重组基底膜及小鼠黑色素瘤高转移株自发性肺转移模型观察了金荞麦提取物对Ｂ１６－ＢＬ６细胞的体外抗侵袭活性和体内抗转移作用；用聚丙烯酰胺凝胶电泳法进一步观察了其对人纤维肉瘤ＨＴ－１０８０细胞Ⅳ型胶原酶的产生及活性的影响；同时用ＷＳＴ法观察了该药的细胞毒性。结果金荞麦提取物在１００ｍｇＬ－１剂量下能明显抑制Ｂ１６－ＢＬ６细胞侵袭；在２００ｍｇｋｇ－１剂量下能有效抑制Ｂ１６－ＢＬ６黑色素瘤细胞在Ｃ５７／ＢＬ６小鼠体内自发性肺转移。该药对Ｂ１６－ＢＬ６和ＨＴ－１０８０细胞无明显细胞毒作用。该药能抑制ＨＴ－１０８０细胞Ⅳ胶原酶的产生，但对酶的活性无明显影响。结论金荞麦提取物具有明显的抗癌侵袭和转移的作用。     

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。     

鲨鱼软骨提取物对C_(57)BL/6J小鼠B_(16)黑色素瘤肺转移的影响

目的：评价鲨鱼软骨提取物（SCAE）对C57BL／6J小鼠B16黑色素瘤肺转移的影响。方法：小鼠经尾静脉接种B16黑色素瘤细胞悬液（5×104／每鼠）后d21，观察SCAE低、中、高剂量及环磷酰胺、生理盐水对照组小鼠肺部转移瘤生长情况。结果：SCAE及环磷酰胺对C57BL／6J小鼠经血道B16黑色素瘤肺转移均有明显的抑制作用，其肺校重及肺转移灶数均低于对照组，但后者主要免疫器官胸腺呈现明显萎缩，而SCAE各剂量组无此毒副作用。     

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。     

Inhibitory effect of MD-Fraction on tumor metastasis: involvement of NK cell activation and suppression of intercellular adhesion molecule (ICAM)-1 expression in lung vascular endothelial cells

The anti-metastatic activity of MD-Fraction extracted from the maitake mushroom (Grifola frondosa) was examined in an experimental murine model of lung metastasis. Intraperitoneal administration of MD-Fraction 2 d before tumor implantation significantly inhibited lung metastasis of colon-26 carcinoma and B16/BL6 melanoma cells. In this model, MD-Fraction enhanced IL-12 production from antigen presenting cells (APCs). MD-Fraction treatment activated NK cells and increased cytotoxicity against YAC-1 and colon-26 carcinoma cells. Furthermore, the depletion of NK cells with anti-asialo GM1 abolished the inhibitory effect of MD-Fraction on lung metastasis of colon-26 cells. Ex vivo, B16/BL6 cell adhesion to LPS-activated murine lung vascular endothelial cells was inhibited by MD-Fraction and anti-intercellular adhesion molecule (ICAM)-1 antibody. These results suggest that MD-Fraction inhibits tumor metastasis by activating NK cells and APCs, and by suppressing of ICAM-1 leading to the inhibition of tumor cell adhesion to vascular endothelial cells.     

Antimetastatic effect of an immunomodulatory arabinomannan extracted from Mycobacterium tuberculosis strain Aoyama B,Z-100, through the production of interleukin-12

In this study, the role of interleukin (IL)-12 on the antimetastatic effect of Z-100 was investigated using wild-type C57BL/6 mice or IL-12p40 knockout (IL-12p40 KO) mice inoculated with highly metastatic B16F10 melanoma. When C57BL/6 mice were inoculated with B16F10 melanoma (2x10(5) cells/mouse i.v.), Z-100 (10 mg/kg i.p.) significantly suppressed the pulmonary metastasis of B16F10 melanoma 14 d after tumor inoculation. On the other hand, the antimetastatic effect of Z-100 was not observed in IL-12p40 KO mice inoculated with B16F10 melanoma. These results indicate that IL-12 is essentially required for the appearance of the antimetastatic effect of Z-100. Since helper T (Th) 2 cell responses have been reported to have a role in tumor metastasis, the regulatory effect of Z-100 on the immune balance of Th1/Th2 cell responses was investigated. In both C57BL/6 mice and IL-12p40 KO mice bearing B16F10 melanoma, Th1 cytokine production (IL-2, interferon-gamma) was significantly suppressed as compared with those in normal mice. On the other hand, Th2 cytokine production (IL-4, IL-10) in these mice was increased. The administration of Z-100 (10 mg/kg i.p.) in C57BL/6 mice bearing B16F10 melanoma improved the balance of Th1/Th2 cell responses from the Th2-dominant state to the normal state. However, the improvement of Th1/Th2 cell responses by Z-100 was not observed in IL-12p40 KO mice bearing the same tumors. In addition, Z-100 significantly increased IL-12 production by macrophages in a concentration-dependent manner, while Z-100 significantly decreased IL-10 production by these cells in vitro. These results suggested that up-regulation of IL-12 production and down-regulation of IL-10 production by Z-100 are related to the improvement of Th1/Th2 cell responses from the Th2-dominant state to the normal state, which resulted in suppression of tumor metastasis.