舒尼替尼衍生物ZH26体内外抗肿瘤作用

目的研究舒尼替尼衍生物ZH26的抗肿瘤作用。方法采用甲基噻唑比色法体外检测ZH26对多种肿瘤细胞株的生长抑制作用;建立S180肉瘤、Colon26结肠癌小鼠肿瘤模型和裸鼠人体肿瘤移植瘤NCI-H460人大细胞肺癌移植瘤、SPC-1肺腺癌移植瘤模型,评价ZH26的体内抗肿瘤活性。结果 ZH26体外对多株肿瘤细胞株有较好的抑制作用,对大多数肿瘤细胞株的IC50值为3.33～5.68μg/mL,与舒尼替尼相当。体内试验表明,30～90 mg/kg ZH26对小鼠S180肉瘤的抑瘤率为40.55%～66.82%;对小鼠Colon26结肠癌的抑瘤率为55.41%～78.14%;对NCI-H460人大细胞肺癌的抑瘤率为48.14%～70.56%;对SPC-1肺腺癌的抑瘤率为42.06%～92.18%。结论 ZH26具有良好的体内外抗肿瘤作用,相关机制尚待进一步研究。     

PARP抑制剂WZ-8体内外抗肿瘤作用研究

目的 研究PARP抑制剂WZ-8对人源性肿瘤细胞增殖及裸鼠移植人结肠癌（HCT116）生长的抑制作用。方法 SRB染色法和裸小鼠移植瘤模型研究WZ-8在体内外抗肿瘤作用。结果 体外WZ-8对多种人肿瘤细胞增殖有明显抑制作用,其中对结肠癌细胞HCT116的半数抑制浓度IC50为（0.21±0.01）mg·mL^-1。体内研究表明WZ-8（20 mg·kg-1）对小鼠移植HCT116肿瘤生长有一定抑制效果,其体内外抗肿瘤活性均高于同类化合物Iniparib。结论 WZ-8在体外对人源性肿瘤细胞增殖有明显抑制作用,在体内可抑制裸鼠移植人结肠癌细胞HCT116的生长。     

全新长春氟宁衍生物体内外药效筛选

目的:对29个全新长春氟宁衍生物进行体内、外药效筛选,以期发现具有开发前景的抗肿瘤化合物.方法:体外筛选采用SRB法,移植入非小细胞肺癌A549裸小鼠模型作为体内药效模型.结果:通过体外IC50结果,结合化学结构,挑选出2个较有优势化合物:SHRll5706和SHR115766,IC50分别为0.05和0.29μmol·L-1.SHR115706能够剂量依赖性的抑制裸鼠移植肿瘤的生长,抗癌疗效与长春氟宁相当;SHR115766亦可抑制肿瘤的生长,但剂量依赖性不明显.SHR115706(80 mg·kg-1),SHR115766(80 mg·kg-1)和长春氟宁(35 mg·kg-1,作为阳性对照)组肿瘤增殖速率分别为47.5%,57.6%和46.5%.SHR115706和SHR115766对裸鼠表现的毒性均明显低于长春氟宁.结论:SHR115706有望成为抗肿瘤新药.     

舒尼替尼与Combretastatin A-4联合用药体外抑制人胃癌SGC-7901细胞增殖作用研究

目的 对舒尼替尼与Combretastatin A-4(CA-4)联合用药的抗肿瘤活性进行研究。方法 SRB法测定比较舒尼替尼、CA-4及联合用药对人胃癌SGC-7901细胞增殖抑制作用。PI染色法、DAPI染色法及Western blot法检测舒尼替尼联合应用CA-4诱导人胃癌SGC-7901细胞凋亡作用。结果 联用舒尼替尼和CA-4在体外能协同抑制人胃癌SGC-7901细胞的增殖。舒尼替尼联用CA-4能够诱导SGC-7901肿瘤细胞凋亡,并且伴随着caspase-3的激活和PARP的裂解。结论 舒尼替尼与CA-4联合用药在体外能协同诱导人胃癌SGC-7901肿瘤细胞增殖。     

多靶点酪氨酸激酶抑制药舒尼替尼

舒尼替尼是一种小分子多靶点酪氨酸激酶抑制药。通过抑制多个受体酪氨酸激酶(RTKs)磷酸化表现出抗肿瘤和抗血管形成活性。相对于更高选择性的激酶抑制剂,它的多靶点协作既具有可以接受的毒性,又大大提高了抗癌活性。2006年1月舒尼替尼获得美国FDA批准用于治疗晚期肾细胞癌(RCC)和服用伊马替尼后疾病出现进展或不耐受伊马替尼的胃肠道间质肿瘤(GIST)。     

盐酸伊立替康脂质体临床前药代动力学、耐受性和药效学研究

目的:研究盐酸伊立替康脂质体与盐酸伊立替康注射液相比,在药代动力学、毒理学和药效方面的改善程度。方法:考察盐酸伊立替康注射液和盐酸伊立替康脂质体分别以1 mg·kg-1经静脉给药在Beagle犬体内的药代动力学;经静脉注射给药对昆明小鼠的最大耐受量(MTD);对裸鼠皮下接种人结肠癌HT-29、人胃癌NCI-N87移植瘤的抑制作用,通过各组药代动力学参数、MTD和相对肿瘤体积的动态变化比较其改善程度。结果:盐酸伊立替康注射液和脂质体在犬体内的t1/2分别为1.50和9.83 h;脂质体的AUC0～t为注射液的95倍;MTD分别为80和160 mg·kg-1;对HT-29和NCI-N87移植瘤模型的最高抑制率分别为54.2%,97.3%和55.4%,98.5%,且给予5 mg·kg-1脂质体的疗效与50 mg·kg-1注射液相当。结论:盐酸伊立替康脂质体在药代动力学、毒理学和对HT-29,NCI-N87的疗效方面明显优于注射液。     

赛特铂抗肿瘤作用的临床前观察

目的观察赛特铂临床前抗肿瘤作用.方法体外观察赛特铂对人卵巢癌细胞A2780,肺腺癌细胞A549,结肠癌细胞HCT8,乳腺癌细胞MCF7和前列腺癌细胞DU145等10株人肿瘤细胞的细胞毒作用.体内观察赛特铂对人卵巢癌A2780,小鼠肉瘤S180和小鼠肝癌HepS的生长抑制作用.结果赛特铂的体外体内作用强度与对照药顺铂相当.对10种肿瘤细胞的半数抑制浓度IC50为0.51～3.29μmol·L-1,平均IC50为(1.44±0.32)μmo1·L-1;体内口服给药明显抑制裸鼠人卵巢癌A2780,小鼠肉瘤S180和小鼠肝癌HepS的生长,抑制率分别为61.3%,57.9%和50.6%.结论赛特铂的体外细胞毒和体内口服抗小鼠肿瘤生长抑制作用均与顺铂相仿.     

基于UPLC-MS技术的紫草素干预结肠癌移植瘤小鼠血清代谢组学研究

研究萘醌类化合物紫草素(shikonin)体外抗肿瘤活性与对SW620结肠癌移植瘤小鼠的抗肿瘤作用及其机制。采用MTT法测定紫草素对4种结直肠癌细胞的半数抑制浓度(IC₅₀);建立人结肠癌SW620裸鼠异体移植瘤模型,通过测定肿瘤体积、抑瘤率观察紫草素在体内的抗肿瘤活性并利用代谢组学技术结合多元统计对小鼠血清进行分析,探讨紫草素抗肿瘤作用机制。结果显示,紫草素能够显著抑制4种结肠癌细胞的增殖,同时在体内具有较高的抗肿瘤活性,在低、高剂量条件下抑瘤率分别达38.35%和42.16%。代谢组学分析共鉴定出包括色氨酸、缬氨酸和甲硫氨酸在内的38个差异代谢物,通路富集发现其主要通过调节SW620结肠癌移植瘤小鼠体内紊乱的氨基酸代谢发挥抗肿瘤作用,其中对色氨酸代谢通路的影响最为显著。本研究表明紫草素能够通过调节氨基酸代谢发挥抗结肠癌作用,为紫草素的进一步开发及临床应用提供理论依据。     

猕猴桃根抗肿瘤作用研究

目的评价猕猴桃根提取物的体外、内抗肿瘤活性。方法猕猴桃根活性成分的分离、纯化采用传统的天然产物化学方法,猕猴桃根先用甲醇回流提取,然后用乙酸乙酯、氯仿、正丁醇萃取。不同组分对体外培养的肿瘤细胞增殖的作用采用磺酰罗丹明B方法;猕猴桃根的体内抗肿瘤作用采用小鼠肿瘤和人肿瘤裸小鼠移植瘤模型评价。结果氯仿提取物对肿瘤细胞增殖抑制作用最强,甲醇提取物次之。体内实验证实氯仿提取物有效地抑制小鼠肝癌模型和人肝癌裸小鼠移植瘤模型的生长,抑制率大概在38.0%。结论猕猴桃根提取物有一定的抗肿瘤作用;其活性成分主要存在于极性较小的组分。     

吡啶类衍生物YLS002体外抗肿瘤作用及小鼠药效学研究

目的 评价吡啶类衍生物YLS002在体内及体外抗肿瘤作用.方法 采用四甲基偶氮唑比色法体外检测YLS002对13种肿瘤细胞株的生长抑制作用,建立Colon 26结肠癌荷瘤小鼠模型和A549肺癌荷瘤裸小鼠模型,评价YLS002在体内的抗肿瘤活性.结果 YLS002具有很强的细胞毒作用,体外IG50约为0.22～6.74μg/mL.静脉注射剂量为0.062 5、0.25和1.0 mg/kg的YLS002对小鼠Colon 26结肠癌瘤的抑制率为52.08%、71.71%和72.69%;对A549肺癌的抑制率为42.62%、49.73%和85.21%.结论 YLS002具有一定的体外抗肿瘤作用,对小鼠Colon 26结肠癌瘤和A549肺癌均有明显抑制作用.     

化合物209对人结直肠癌细胞HT-29的体内和体外作用(英文)

化合物209是一个新合成的氨基二硫代甲酸酯类化合物,它在体外水平可以抑制肿瘤细胞的增殖,但是化合物209的体内抗肿瘤作用及其抗肿瘤机制并不明确。本文探究了化合物209对人结直肠癌细胞HT-29的作用并初步探讨了相关机制。体外研究表明,化合物209可以显著抑制HT-29细胞的增殖;体内研究结果表明,化合物209可以显著抑制裸鼠HT-29移植瘤的生长,但是对裸鼠体重和白细胞无影响。流式细胞分析实验结果表明,化合物209可将HT-29细胞阻滞于细胞周期的G1期。同时,化合物209能上调体外培养HT-29细胞中p27,cyclin E,CDK2,cyclin D1和CDK4的表达。在体内瘤组织中上述蛋白表达情况与体外实验结果一致。这些结果说明,化合物209具有较好的抗肿瘤活性,其抗肿瘤作用与细胞周期阻滞及其相关蛋白的表达变化有关。     

Binding and internalization of NGR-peptide-targeted liposomal doxorubicin (TVT-DOX) in CD13-expressing cells and its antitumor effects

In an effort to develop new agents and molecular targets for the treatment of cancer, aspargine-glycine-arginine (NGR)-targeted liposomal doxorubicin (TVT-DOX) is being studied. The NGR peptide on the surface of liposomal doxorubicin (DOX) targets an aminopeptidase N (CD13) isoform, specific to the tumor neovasculature, making it a promising strategy. To further understand the molecular mechanisms of action, we investigated cell binding, kinetics of internalization as well as cytotoxicity of TVT-DOX in vitro. We demonstrate the specific binding of TVT-DOX to CD13-expressing endothelial [human umbilical vein endothelial cells (HUVEC) and Kaposi sarcoma-derived endothelial cells (SLK)] and tumor (fibrosarcoma, HT-1080) cells in vitro. Following binding, the drug was shown to internalize through the endosomal pathway, eventually leading to the localization of doxorubicin in cell nuclei. TVT-DOX showed selective toxicity toward CD13-expressing HUVEC, sparing the CD13-negative colon-cancer cells, HT-29. Additionally, the nontargeted counterpart of TVT-DOX, Caelyx, was less cytotoxic to the CD13-positive HUVECs demonstrating the advantages of NGR targeting in vitro. The antitumor activity of TVT-DOX was tested in nude mice bearing human prostate-cancer xenografts (PC3). A significant growth inhibition (up to 60%) of PC3 tumors in vivo was observed. Reduction of tumor vasculature following treatment with TVT-DOX was also apparent. We further compared the efficacies of TVT-DOX and free doxorubicin in the DOX-resistant colon-cancer model, HCT-116, and observed the more pronounced antitumor effects of the TVT-DOX formulation over free DOX. The potential utility of TVT-DOX in a variety of vascularized solid tumors is promising.     

Antitumor effects of recombinant human prolactin in human adenocarcinoma-bearing SCID mice with human NK cell xenograft

To survey the immune regulatory function of recombinant human prolactin (rhPRL) and its potential application in adoptive immunotherapy, CB17-SCID mice were loaded with human colon adenocarcinoma HT-29 cells (5 x 10(5) cells/mouse, i.p.) 24 h before adoptive transfer with the purified human NK cells followed by rhPRL injection (10 mug/mouse, every other day for a total of 10 injections). Upon analysis, rhPRL did not exert any direct inhibitory effects on HT-29 cells but slightly improved the tumor cell growth both in vitro and in vivo. After SCID mice were reconstituted with human NK cells, rhPRL improved the antitumor effects of human NK cells in HT-29-bearing SCID mice, showing a prolonged survival from 70.4 to 112.1 days, and the increased survival rate from all died to 40% survival for more than 160 days. rhPRL improved the proliferation of human NK cells with or without PHA stimulation. rhPRL also directly enhanced the cytotoxicity of human NK cells against HT-29 tumor cells in 4-h coculture. The supernatant of rhPRL-stimulating NK cells inhibited the proliferation of HT-29 cells through, at least partly, the interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) in the supernatant. Thus, rhPRL administration in HT-29 tumor-bearing SCID mice promotes the antitumor effects of adoptively transferred NK cells.     

姜黄素与5-氟尿嘧啶联用对人结肠癌HT-29细胞增殖的影响

目的　体外观察 5 氟尿嘧啶与姜黄素联用对人结肠癌HT 2 9细胞增殖的相互作用。方法　采用MTT法观察不同浓度姜黄素、5 氟尿嘧啶单独或联合应用对HT 2 9细胞生长的抑制作用 ,并利用中效原理判定两药合用的效果。结果　两种药物单独应用时 ,对HT 2 9细胞的抑制作用呈明显的剂量效应关系 ;对于HT 2 9细胞 ,姜黄素的中位抑制浓度为(32± 11) μmol·L- 1,5 氟尿嘧啶的中位抑制浓度为(10 4 0± 4 5 6 ) μmol·L- 1。应用中效原理分析显示 ,两药在联合应用时为协同效应 ,并且在不同的药物浓度配比下呈相同的趋势。增大姜黄素的用量可在更宽的效应范围内获得两种药物的协同效应。结论姜黄素与 5 氟尿嘧啶在联合应用时的相互作用为协同效应 ,本结果对于结肠癌的治疗具有一定的临床意义。     

舒尼替尼对EGFR TKI抵抗的A549细胞株增殖抑制作用及机制研究

目的探讨舒尼替尼(sunitinib)体外对表皮生长因子受体酪氨酸激酶抑制剂(EGFR TKI)耐药的非小细胞肺癌A549细胞的生长抑制作用及其机制。方法 MTT法检测sunitinib对A549细胞的生长抑制作用;荧光显微镜观察sunitinib诱导的细胞凋亡;流式细胞术检测sunitinib作用后细胞周期变化;Western blot检测sunitinib作用后Bcl-2蛋白水平的变化。结果在0.4～12.8μmol.L-1浓度范围内,sunitinib抑制A549细胞增殖,且具有浓度和时间依赖性,24、48、72 h的半数抑制浓度(IC50)分别为(7.34±0.76)、(5.54±0.62)、(3.68±0.53)μmol.L-1。荧光显微镜观察发现sunitinib能够诱导细胞出现核固缩、体积缩小等凋亡形态学改变。细胞周期显示sunitinib将A549细胞阻滞于G0/G1期。Western blot显示sunitinib能够降低Bcl-2蛋白的表达水平,且随着sunitinib浓度的升高,Bcl-2蛋白的表达水平逐渐降低。结论 sunitinib能够抑制EGFR TKI抵抗的A549细胞生长,并具有浓度和时间依赖性,其抗肿瘤的机制可能与诱导细胞周期阻滞和下调Bcl-2的表达有关。     

甲氧基咔唑类化合物的合成及其体外抗肿瘤活性

目的设计合成新型甲氧基咔唑类抗肿瘤化合物。方法将4，5-二甲氧基-2-溴硝基苯与甲氧基碘代苯化合物在铜粉催化下经Ullmann反应制得甲氧基-硝基联苯化合物，再经亚磷酸三乙酯还原得三种甲氧基咔唑。对9-位氮进行修饰，合成了16个衍生物。利用核磁共振、质谱、红外光谱和元素分析进行了结构确认。结果体外初步抗肿瘤活性测试结果显示，目标物4_c，5_a，5_b，5_g，5_h，5_i，5_I，5n和5_p对人结肠癌HT-29细胞的IC₅₀值分别12.1，10.6，8.1，3.1，4.4，10.1及9.2 μmol·L^-1。目标物4_a对人口腔上皮癌KB细胞的IC₅₀值为17.7 μmol·L^-1。结论部分目标化合物对HT-29及KB肿瘤细胞具有较好的抑制作用。     

4-(甲基苯胺基)-3-氰基喹啉类衍生物的 合成及抗肿瘤活性

目的 设计合成4-(甲基苯胺基)-3-氰基喹啉类衍生物,并对其体外抗肿瘤活性进行初步评价。 方法 以氰乙酸乙酯为起始原料,经多步反应合成目标化合物。采用MTT法,以吉非替尼（gefitinib）为阳性对照药,以A549、HT-29和MDA-MB-231为测试细胞株对目标化合物的抗肿瘤活性进行了评价。 结果 合成了18个新化合物,经1H-NMR、MS和IR确认其结构。体外活性测试结果显示,多数化合物可在较低浓度抑制肿瘤细胞增殖,其中的Ⅶ2、Ⅶ3、Ⅶ6、Ⅶ12、Ⅶ13、Ⅶ15 和 Ⅶ16共7个化合物有显著的抗增殖活性,优于阳性对照药吉非替尼。 结论 体外活性实验表明：4-(甲基苯胺基)-3-氰基喹啉类衍生物作为新型的酪氨酸激酶抑制剂,其构效关系值得进一步研究。     

Synthesis, antitumor, and antibacterial activity of bis[4,5-diarylimidazol-2-ylidene]methane derivatives

Cationic [bis(1,3-diethyl-4,5-diarylimidazol-2-ylidene)]Au(I) bromide complexes have demonstrated considerable potential as new antitumor agents. In order to investigate whether the gold is crucial for the antitumor activity, the imidazole ligands were connected by a methylene bridge. Biological evaluation revealed that bis[1,3-diethyl-4,5-diarylimidazol-2-ylidene]methane compounds exhibited growth inhibition effects against mammary (MCF-7 and MDA-MB 231) and colon (HT-29) carcinoma cell lines. In comparison with gold complexes, the methylene derivatives showed drastically reduced cell growth inhibitory properties. However, the growth of bacteria was significantly inhibited by bis[1,3-diethyl-4,5-bis(4-methoxyphenyl)imidazol-2-ylidene]methane dibromide (4) and opens a new application of this compound type.     

Inhibition of self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells by scutellarin via Hedgehog signaling pathway

OBJECTIVE To investigate the inhibitory effect of scutellarin on the self-renewal and differentiation of HT-29 cells-derived cancer stem-like cells(HT-29 CSC) in vitro and in vivo, and to explore its mechanism. METHODS The effect of scutellarin on the growth of HT-29 CSC was determined by 3 D Culture assay. The effect of scutellarin on growth and transformation of HT-29 CSC was probed by soft agar colony formation assay. The effect of scutellarin on the differentiation of HT-29 CSC was determined by serum induction differentiation assay in vitro. The effects of scutellarin on the expressions of marker gene Lgr5, target gene c-Myc, proliferation gene CK20 and Nanog gene were measured by quantitative real-time RT-PCR. Investigate the effect of scutellarin on the expression of c-Myc, Gli1, and Lgr5 protein by Western blotting. A subcutaneous xenograft model of colon cancer in nude mice was established and administered by intraperitoneal injection. The change of body weight and tumor size of nude mice were observed every two days. Investigate the effects of scutellarin on the growth of xenograft tumors in nude mice. The expression of CD133, Lgr5, Gli1,Ptch1, c-Myc, Ki67, CK20, Nanog gene in tumors were measured by quantitative real-time RT-PCR. The expression of c-Myc, Gli1, Lgr5, CD133, Ki67 protein were measured by Western blotting. RESULTS Scutellarin can inhibit the growth of HT-29 CSC in 3 D culture. Compared with the solvent control group, scutellarin can significantly inhibit the growth and transformation and differentiation of HT-29 CSC in vitro(P<0.01). The expression levels of marker genes Lgr5, target gene c-Myc, proliferation gene CK20 and Nanog in HT-29 CSC were down-regulated by scutellarin. Scutellarin can reduce the expression of c-Myc, Gli1, and Lgr5 protein in HT-29 CSC.Scutellarin can inhibit the growth of colon cancer xenografts, lower CD133, Lgr5, Gli1, Ptch1, c-Myc, Ki67, CK20, and Nanog mRNA level of xenograft tumors, reduce the expression of c-Myc, Gli1, Lgr5, CD133, and Ki67 protein of xenograft tumors in nude mice. CONCLUSION Scutellarin,which is the main component of scutellaria barbata, can inhibit the differentiation of HT-29 CSC and the mechanism is to inhibit the activity of Hedgehog signaling pathway.     

Application of a tissue culture microtiter test for the detection of cytotoxic agents from natural products

A method is described by which the growth inhibitory effects of cytotoxic compounds and fermentation broth cultures on adherent tumor cell lines can be quantitated. Cells are seeded into 96-well microtiter plates and 16 hours later the test compounds or broths are added to the wells. Cell growth is measured after three days (B16 mouse melanoma cells) or six days (HT-29, human colon carcinoma cells) by first fixing adherent cells, staining with Giemsa stain, washing away excess stain, then solubilizing stained cells with HCl. Absorbance is determined using a microELISA spectrophotometer and the data are transferred to and analyzed by a computer. The assay is rapid and reproducible and can be used to identify fermentation broths with cytotoxic components. Addition of DNA into the assay mixture (cells plus compound) inhibits the cytotoxic activities of certain DNA-reactive agents. The results of this study demonstrate the application of this assay system for primary and secondary evaluation of fermentation broths for in vitro antitumor activity.