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.     

肿瘤细胞裂解物抗小鼠B16F10黑色素瘤的作用研究

反复冻融B16F10肿瘤细胞制备裂解物,以白喉毒素(Diphtheria toxin,DT)为载体,OK432和来源于结核分枝杆菌(Mycobacterium tuberculosis)热休克蛋白70(HSP70)第407-426(mHSP70407～426,M)的两段串联重复序列M2为佐剂,制备了肿瘤细胞疫苗B16F10-DT-M2-OK432(BDTMOK),探讨其能否抑制小鼠B16黑色素瘤,并且对其抗肿瘤的作用机理进行部分探讨。以制备的BDTMOK免疫C57BL/6小鼠,分别检测体液免疫应答和细胞免疫应答。通过ELISA法,从血清中检测到高滴度的抗B16肿瘤细胞裂解物(B16 tumor cell lysate,B16TCL)类抗体。淋巴细胞增殖实验的结果显示,BDTMOK的免疫能够有效的刺激脾淋巴细胞的增殖。预防结合治疗性实验的结果显示,BDTMOK激发的免疫应答对于B16肿瘤攻击起到有效的保护作用,与PBS阴性对照组比较,皮下注射BDTMOK可以延长皮下移植瘤发生的潜伏期(P<0.05),并且平均瘤重显著降低(P<0.05);抑制了小鼠皮内肿瘤模型中的血管新生(P<0.01)。疫苗BDTMOK能有效的抑制小鼠B16黑色素瘤的生长。     

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。     

四种含药血清对小鼠B16黑素瘤细胞黑素合成及酪氨酸酶活性的影响

目的研究传统医学文献中专治白癜风的常用药物诃子、余甘子、柴胡及药群组合(含木香、胡椒、干姜等)的药物血清对小鼠B16黑素瘤细胞黑素合成及酪氨酸酶活性的影响。方法采用"通法"制备上述药物高、低不同剂量的大鼠含药血清及不含药物的空白血清,以体外培养的小鼠B16黑素瘤细胞为模型,采用MTT法测定各组受试血清对B16细胞增殖活性的影响;NaOH细胞裂解法测定黑素合成量,L-DOPA氧化法测定对酪氨酸酶活性的影响。结果与正常对照组(含10%小牛血清)相比,除组合低剂量组外,其他受试血清均不影响小鼠B16细胞的增殖活性(P>0.05);与空白血清组相比,柴胡和药群组的高、低剂量组含药血清均对B16黑素瘤细胞黑素合成有促进作用(P<0.05,P<0.01),而诃子低剂量、牛柑子高剂量组含药血清具有促进黑素合成的作用(P<0.05,P<0.01);与空白血清组相比,四种受试药物的高低剂量组含药血清对小鼠B16细胞酪氨酸酶活性均有激活作用(P<0.05,P<0.01)。结论四种受试药物含药血清除组合低剂量组外,均在不影响小鼠B16细胞增殖活性的基础上,表现出不同程度的促进其黑素合成的作用,此作用与提高酪氨酸酶活性有一定关系。     

羟基喜树碱对胃癌细胞凋亡和细胞周期的影响

目的 探讨羟基喜树碱诱导胃癌细胞凋亡与细胞周期变化的关系.方法 用MTT法测定不同浓度的HCPT对胃癌细胞的生长抑制作用,用流式细胞仪测定一定浓度下不同作用时间细胞凋亡率和细胞周期的变化.结果 在3.125～50.0 μg/ml浓度范围内药物对细胞的抑制率随浓度的增加而增加;在相同浓度下,随时间的延长细胞的凋亡率也逐渐增加,0、12、24、48h的凋亡率为2.8%、8.5%、10.2%,13.3%,在24h内S期细胞减少而G0/G1细胞增多,第二个24h S期和G2/M的阻滞.结论羟基喜树碱可诱导胃癌细胞凋亡,其细胞凋亡与细胞周期分布有关.     

鱿鱼皮胶原蛋白多肽对B16黑素瘤细胞黑素合成的影响

目的研究不同分子量鱿鱼皮胶原蛋白多肽SP1(Mr>10000u)、SP2(6000u相似文献   

甘草查尔酮A抑制小鼠黑色素瘤B16F10细胞增殖机制研究

目的研究甘草查尔酮A抑制B16F10细胞增殖机制。方法 SRB法检测甘草查尔酮A对B16F10细胞增殖影响,Giemsa染色法观察细胞形态变化,比色法检测B16F10细胞内、外黑色素含量,Annexin V-FITC/PI双染检测细胞凋亡率,流式细胞术测定细胞周期分布,Q-PCR法检测细胞凋亡相关基因B淋巴细胞-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞周期蛋白(Cyclin E2)和细胞周期蛋白依赖性激酶-2(CDK2)的mRNA表达。结果甘草查尔酮A能有效抑制B16F10细胞增殖,呈现浓度依赖性和时间依赖性;随药物浓度增加,细胞增殖速度降低,细胞形态由树突状变为固缩圆球状,并伴有黑色素颗粒物出现,且细胞内、外黑色素含量呈浓度依赖性增加趋势,甘草查尔酮A能使细胞阻滞在G1期,在低浓度时,诱导细胞分化,高浓度时,诱导细胞凋亡;同时,甘草查尔酮A下调凋亡相关蛋白Bcl-2/Bax比率,抑制周期相关蛋白Cyclin E2、CDK2的mRNA表达。结论甘草查尔酮A抑制B16F10细胞增殖机制可能是通过使B16F10细胞G1期阻滞,进而诱导细胞分化和凋亡。     

青蒿琥酯对黑素瘤A875细胞增殖和凋亡的影响及机制研究

目的观察青蒿琥酯对人皮肤恶性黑素瘤A875细胞增殖和凋亡的影响,并探讨相关机制。方法体外培养A875细胞,分别给予不同浓度的青蒿琥酯(10~80μg/ml)作用细胞48 h,MTT法检测抑制率,流式细胞术检测凋亡率,Western blot法检测Caspase-3、Caspase-8、Caspase-9的蛋白表达水平变化。结果青蒿琥酯对A875细胞有明显的生长抑制和诱导凋亡的作用(P<0.01),呈剂量依赖性;随药物浓度的增高,Caspase-3、Caspase-8、Caspase-9的蛋白表达量逐渐升高(P<0.05)。结论青蒿琥酯可以通过死亡受体途径和线粒体途径诱导A875细胞的凋亡,从而抑制细胞增殖。     

肿瘤特异性凋亡基因对人黑素瘤细胞凋亡诱导作用的机制研究

为研究肿瘤特异性凋亡基因(apoptin gene)在诱导人黑素瘤细胞A375发生凋亡时，c-Jun氨基末端激酶(JNK)信号通路在细胞发生凋亡中的作用，用含有apoptin基因的真核表达载体瞬间转染体外培养的人黑素瘤细胞A375；采用流式细胞仪、蛋白印迹法(Western blot)、锥虫蓝染色法及RT-PCR等方法研究其在不同时间对人黑素瘤细胞A375诱导凋亡的作用。结果表明，apoptin基因瞬间转染的人黑素瘤细胞A375可出现明显的细胞凋亡；而且凋亡细胞的数量与apoptin的转染时间有关；凋亡细胞内磷酸化型JNK蛋白随凋亡细胞数量的增加而增多，而非磷酸化型JNK蛋白表达无明显变化。结论：JNK信号通路的活化可能在apoptin诱导肿瘤细胞凋亡过程中发挥作用。     

10-羟基喜树碱诱导人肝癌细胞SMMC-7721凋亡的探讨

目的：为筛选新的凋亡诱导剂，并对其机制进行研究。方法：用10－羟基喜树碱（10－hydroxycamp-tothecin,10-HCPT）以不同终浓度（0－150μm），不同作用时间（0－24h)诱导体外培养的SMMC－7721人肝癌细胞，按设计时间（24－140h）收集悬浮细胞，贴壁细胞和对照组进行各组间凋亡率，残存率、克服形成率观察。结果：当10－HCPT终浓度＞2．0μm时，部分细胞出现典型凋亡特征，随浓度及作用时间增加，凋亡率也随之升高，残存率下降，克隆形成率明显下，当10＝HCPT浓度＞50μm时，大量凋亡细胞中出现坏死，结论：10－HCPT能诱导SMMC－7721细胞凋亡，其效果与剂量和时间密切相关。     

Evaluation of anti-metastatic potential of Cisplatin polymeric nanocarriers on B16F10 melanoma cells

Nanoparticles are being increasingly used in the field of cancer treatment due to their unique properties and advantages. The aim of the present research work was to prepare and characterize a polymeric albumin nanosystem for Cisplatin and evaluate its in-vitro efficacy against B16F10 melanoma. The developed nanoparticles were almost spherical in shape with a particle size in the range of 150–300 nm, low polydispersity values and about 80% drug entrapment efficiency. Albumin nanocarriers sustained the release of Cisplatin for more than 48 h, suggesting the reduction in dosing schedule for this drug. The results from in-vitro cell line studies indicated the dose dependent cytotoxic potential of drug loaded albumin nanoparticles, their potential to inhibit cell proliferation and induce morphological changes. In addition, these nanoparticles exhibited superiority to Cisplatin in hampering the cell migration. Developed nanoparticles caused cell cycle arrest along with time and concentration dependent cellular uptake in B16F10 cell line. These results signify that the prepared Cisplatin albumin nanoparticles could serve as a promising approach for B16F10 melanoma treatment.     

Inhibitory effect of antidepressants on B16F10 melanoma tumor growth

BackgroundAntidepressant drugs, like fluoxetine, a selective serotonin reuptake inhibitor, desipramine, a nonselective noradrenaline reuptake inhibitor, and mirtazapine, an antagonist of noradrenaline α₂ auto- and heteroreceptors, are widely used for the treatment of depressive symptoms in cancer patients. Since these antidepressants have different activities targeting the immune system, they might also modulate tumor growth in cancer patients.MethodsIn the present study, we investigated the effects of administration of antidepressant drugs: fluoxetine, desipramine and mirtazapine on B16F10 melanoma tumor growth. These drugs were administered intraperitoneally (ip) for 17 days after subcutaneous injection of B16F10 melanoma cells to male C57BL/6J mice.ResultsFluoxetine significantly inhibited melanoma solid tumor growth and desipramine tended to decrease this parameter whereas mirtazapine had no effect.ConclusionThe inhibitory effect of fluoxetine on melanoma growth was associated with an increased mitogen-induced T cell proliferation which may at least partly participate in the mechanism of the antitumor effect of this antidepressant. It appears that the inhibitory effect of fluoxetine on tumor growth is not related with changes in cytokine levels except for IL-10.     

Suppression of the migration and invasion is mediated by triptolide in B16F10 mouse melanoma cells through the NF‐kappaB‐dependent pathway

Melanoma cancer is one of the major causes of death in humans worldwide. Triptolide is one of the active components of Tripterygium wilfordii Hook F, and has biological activities including induced cell cycle arrest and induction of apoptosis but its antimetastatic effects on murine melanoma cells have not yet been elucidated. Herein, we investigated the effect of triptolide on the inhibition of migration and invasion and possible associated signal pathways in B16F10 murine melanoma cancer cells. Wound healing assay and Matrigel Cell Migration Assay and Invasion System demonstrated that triptolide marked inhibiting the migration and invasion of B16F10 cells. Gelatin zymography assay demonstrated that triptolide significantly inhibited the activities of matrix metalloproteinases‐2 (MMP‐2). Western blotting showed that triptolide markedly reduced CXCR4, SOS1, GRB2, p‐ERK, FAK, p‐AKT, Rho A, p‐JNK, NF‐κB, MMP‐9, and MMP‐2 but increased PI3K and p‐p38 and COX2 after compared to the untreated (control) cells. Real time PCR indicated that triptolide inhibited the gene expression of MMP‐2, FAK, ROCK‐1, and NF‐κB but did not significantly affect TIMP‐1 and ‐2 gene expression in B16F10 cells in vitro. EMSA assay also showed that triptolide inhibited NF‐κB DNA binding in a dose‐dependent manner. Confocal laser microscopy examination also confirmed that triptolide inhibited the expression of NF‐κB in B16F10 cells. Taken together, we suggest that triptolide inhibited B16F10 cell migration and invasion via the inhibition of NF‐κB expression then led to suppress MMP‐2 and ‐9 expressions. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1974–1984, 2016.     

Capsaicin induced apoptosis of B16-F10 melanoma cells through down-regulation of Bcl-2.

Capsaicin (8-methyl-N-vanillyl-6-nonenamide), a pungent ingredient of hot chili peppers, has been reported to possess substantial anticarcinogenic and antimutagenic activities. In the present study, we investigated the effect of capsaicin on induction of apoptosis in highly metastatic B16-F10 murine melanoma cells. Capsaicin inhibited growth of B16-F10 cells in a concentration-dependent manner. Proapoptotic effect of capsaicin was evidenced by nuclear condensation, internucleosomal DNA fragmentation, in situ terminal nick-end labeling of fragmented DNA (TUNEL), and an increased sub G1 fraction. Treatment of B16-F10 cells with capsaicin caused release of mitochondrial cytochrome c, activation of caspase-3, and cleavage of poly (ADP-ribose) polymerase in a dose-dependent manner. Furthermore, Bcl-2 expression in the B16-F10 cells was slightly down-regulated by capsaicin treatment. In contrast, there were no alterations in the levels of Bax in capsaicin-treated cells. Collectively, these findings indicate that capsaicin-induces apoptosis of B16-F10 melanoma cells via down-regulation the Bcl-2.     

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 .     

Effects of benzoylphenyl ureas on growth of B16 melanoma cells in vitro and in vivo

Summary New analogs of diflubenzuron, a benzoylphenyl urea, are tested on their in vitro cytostatic activity against B16 melanoma cells. The following structure-activity relationship was established: substitution by a hydroxylated function at the ortho, meta or para position or by a dimethylamino function at the ortho position of the benzoyl moiety appeared to be necessary for cytostatic activity in vitro. Acetoxy functions at the ortho position or hydroxy functions at the para position of the aniline ring resulted also in active compounds. A number of these benzoylphenyl ureas are selected for in vivo evaluation of antitumor activity on B16 melanoma growing s.c. Although many of the tested benzoylphenyl ureas delayed tumor growth during the first ten days of drug treatment, only a few increased animal life span. The best results (%T/C) were obtained with compounds 5 (127%), 7 (147%), 13 (135%) and 16 (135%), which all have hydroxylated functions in the benzoyl moiety.     

大豆甙元对小鼠B16黑色素瘤细胞的分化诱导作用

大豆甙元在10～40μg·ml~(-1)浓度范围内,能够明显抑制B16细胞的增殖,受药物作用4d的B16细胞,克隆形成能力及体内成瘤能力明显降低,大豆甙元在抑制B16细胞增殖的同时,促进黑色素的生成,且对B16细胞形态具有明显的影响,低浓度时促使细胞平行排列,当浓度增加时,形成网状结构,黑色素颗粒明显增多。