3种选择性环氧合酶-2抑制剂对胰腺癌生长的影响

目的 比较3种不同选择性的环氧合酶-2(COX-2)抑制剂--美格昔康、塞来昔布、罗非昔布对人胰腺癌细胞生长及凋亡的影响，观察罗非昔布对人裸鼠胰腺癌移植瘤生长的抑制作用。方法采用3H-胞腺嘧啶核苷掺入，了解细胞DNA合成；用免疫组化检测增殖细胞核抗原(proliferating cell nuclear antigen，PCNA)及细胞COX-2的表达；用TUNEL染色法检测细胞凋亡。建立裸鼠胰腺癌移植瘤模型，给予罗非昔布8周，观察肿瘤大小、肿瘤组织的COX-2及PXNA的表达情况。结果 3种COX-2抑制剂均能抑制体外培养的胰腺癌细胞株BxPC-3的3H-胸腺嘧啶核苷掺入，其抑制效应与药物浓度呈显著正相关。COX-2抑制剂的选择性越高，对胰腺癌细胞生长的抑制作用越强。COX-2抑制剂均能诱导胰腺癌细胞的凋亡、降低胰腺癌细胞的COX-2、PCNA的表达。罗非昔布对裸鼠胰腺癌移植瘤的抑瘤率为87．7％；肿瘤组织的COX-2、PCNA表达均较对照组明显下降。结论 3种选择性COX-2抑制剂均能抑制人胰腺癌细胞的生长，并诱导其凋亡，对COX-2的选择性越高，对胰腺癌的抑制作用越强。罗非昔布可显著抑制裸鼠胰腺癌移植瘤的生长。     

环氧化酶-2抑制药对肺癌细胞和骨肉瘤细胞生长的抑制作用

目的观察选择性环氧化酶-2（COX-2）抑制药塞来昔布、罗非昔布对人骨肉瘤细胞株（HOS-8603）、肺癌细胞株（A-549）的作用以及罗非昔布、塞来昔布对两种细胞抑制效应的比较。方法MTT法测定体外培养的细胞株HOS-8603、A-549对不同浓度的罗非昔布和塞来昔布的敏感性，流式细胞仪分析细胞增殖周期的影响及凋亡。结果塞来昔布、罗非昔布均可明显抑制HOS 8603、A-549细胞的生长（P＜0.05），抑制效应呈现浓度依赖型和时间依赖型。HOS-8603和A-549细胞分裂阻滞在G2/M期，出现凋亡峰，塞来昔布对HOS 8603的作用强于罗非昔布。结论COX-2抑制药塞来昔布尼罗非昔布对骨肉瘤细胞、肺癌细胞的生长有抑制作用，有望成为抗肿瘤新药。     

帕瑞昔布对人结肠癌SW1116细胞裸鼠移植瘤生长的影响及安全性观察

目的观察帕瑞昔布对人结肠癌SW1116细胞裸鼠皮下移植瘤生长的抑制作用及安全性。方法建立人结肠癌SW1116细胞裸鼠皮下移植瘤模型。将24只BALB/C裸鼠随机分为对照组、帕瑞昔布组和5-Fu组。记录各组裸鼠的体重和瘤体积变化,用药30 d后取瘤及脾脏,计算脾脏指数、抑瘤率,检测血清ALT和BUN,并用透射电镜观察瘤组织的形态学变化。结果帕瑞昔布组未见明显的药物相关毒副反应。帕瑞昔布能抑制SW11 16细胞裸鼠皮下移植瘤的生长。在干预前18 d,帕瑞昔布组的肿瘤抑制率逐渐增加,但随着药物干预时间的延长,抑制率却逐渐降低。结论裸鼠体内短期应用帕瑞昔布(15~18 d)是安全的,并能抑制SW1116细胞皮下移植瘤的生长。     

奥曲肽抑制人卵巢癌细胞增殖

目的 探讨生长抑素类似物奥曲肽在体内外对人卵巢癌细胞株SKOV3生长的影响.方法 Mtt比色法观察体外奥曲肽对人卵巢癌细胞株SKOV3的生长抑制作用.建立人卵巢癌细胞株SKOV3裸鼠移植瘤模型,随机分成四组,分别给予生理盐水(A组)、奥曲肽(B组)、顺铂(C组)及两药联用(D组)处理,处死裸鼠后采用免疫组织化学法检测肿瘤标本血管内皮生长因子(VEGF)的表达.结果 奥曲肽可抑制体外SKOV3细胞生长,且其作用呈时间、浓度依赖性.各用药组瘤重、瘤体积、VEGF的表达均低于A组(P<0.05).结论 奥曲肽能抑制体内外人卵巢癌细胞株SKOV3生长,对肿瘤VEGF表达的抑制可能为其抑瘤机制之一.     

奥曲肽对食管癌细胞DNA合成及端粒酶活性的影响

目的 研究奥曲肽对食管癌细胞DNA合成及端粒酶活性的影响。方法 将不同浓度的奥曲肽加入食管癌细胞株Eca9706，3^H-胸腺嘧啶核苷(3^H—TdR)掺入实验测定对DNA合成的影响，显微镜下观察细胞形态变化，采用TRAP PCR-ELISA法对端粒酶的活性进行检测。结果 奥曲肽对Eca9706细胞的DNA合成有明显抑制作用，镜下可见明显的细胞病变，细胞变圆、变小、脱落，端粒酶的活性显受抑制。结论 奥曲肽能明显抑制食管癌细胞株Eca9706的DNA合成及端粒酶活性。     

奥曲肽联合顺铂对人胃癌细胞株成瘤的影响

<正>奥曲肽和顺铂对MGC-803细胞在体外都有抑制生长和诱导凋亡的作用。本实验是通过观察奥曲肽和顺铂对人胃癌MGC-803裸鼠种植瘤的成瘤情况及生长的影响,进一步研究奥曲肽联合顺铂对胃癌生长的     

昔布类药物的临床安全性

昔布类药物即高选择性COX-2抑制剂,通过选择性抑制COX-2,阻断花生四烯酸合成前列腺素而发挥抗炎镇痛作用,可减少传统NSAID药物消化系统的不良反应,主要品种有罗非昔布、塞来昔布、伐地昔布、帕瑞昔布、依托昔布及卢米昔布等。其中罗非昔布(万络)在2004年9月由于心血管不良反     

奥曲肽联合顺铂对人胃癌细胞株MGC-803生长影响的研究

目的 研究奥曲肽联合顺铂对人胃癌细胞株MGC-803增殖、凋亡的影响及其可能的作用机制.方法 利用不同浓度的奥曲肽、顺铂以及一定浓度的奥曲肽联合顺铂作用于体外培养的人胃癌细胞株MGC-803,应用MTT 法分析细胞增殖的抑制作用,流式细胞仪检测细胞周期及凋亡率.结果 奥曲肽及顺铂对MGC-803细胞的生长增殖产生抑制作用,并具有量效、时效及饱和性;大多数细胞被阻滞于G0/G1期,S期和G2/M期细胞数明显减少,细胞分裂受到抑制,凋亡率增加.结论 奥曲肽联合顺铂抑瘤率及细胞凋亡率显著高于单用奥曲肽或顺铂治疗组,表明奥曲肽和顺铂有协同作用,这为临床奥曲肽联合顺铂治疗胃癌提供了理论基础.     

奥曲肽对人卵巢癌耐顺铂细胞株SKOV3/DDP移植瘤增殖的影响

目的探讨奥曲肽对人卵巢癌耐顺铂细胞株SKOV3/DDP裸鼠移植瘤生长的影响。方法将SKOV3/DDP细胞接种雌性BALB/c-nu/nu裸鼠右腋前皮下构建移植瘤动物模型,成瘤后随机分成4组。奥曲肽组(A):奥曲肽100μg.kg-1.d-1,用药28 d;顺铂组(B):顺铂4 mg.kg-1.d-1,成瘤后第1、8、15、22天用药;联合组(C):奥曲肽和顺铂联用;对照组(D):生理盐水对照。用药4周后处死裸鼠,取肿瘤标本计算瘤重和瘤体积,并采用免疫组织化学法检测瘤块中生长抑素受体2(SSTR2)和表皮生长因子受体(EGFR)的表达。结果各用药组肿瘤标本的瘤重、瘤体积均低于D组,C组的瘤重、瘤体积低于A、B组(P<0.01)。各组移植瘤细胞均有SSTR2表达;EGFR表达量D组显著高于其他各组(P<0.01),C组及A组EGFR表达量低于其他两组(P<0.01)。结论奥曲肽能抑制人卵巢癌耐顺铂细胞株SKOV3/DDP的裸鼠移植瘤的生长,并与顺铂有协同作用。     

特异性环氧化酶 Ⅱ抑制药——罗非昔布

综述非甾体抗炎药——罗非昔布的药理作用特点和临床应用.研究表明,罗非昔布可有效地抑制环氧化酶 Ⅱ,能够减少脂多糖诱导的前列腺素E2的合成,而对血小板或胃粘膜中的血栓素B2的合成无明显影响;临床应用显示,罗非昔布对骨关节炎及剧痛的疗效较好,副作用较小.     

The anti-proliferative potency of celecoxib is not a class effect of coxibs

Celecoxib, a COX-2 (cyclooxygenase-2)-selective inhibitor (coxib), is the only NSAID (non-steroidal anti-inflammatory drug) that has been approved for adjuvant treatment of patients with familial adenomatous polyposis. To investigate if the anti-proliferative effect of celecoxib extends to other coxibs, we compared the anti-proliferative potency of all coxibs currently available (celecoxib, rofecoxib, etoricoxib, valdecoxib, lumiracoxib). Additionally, we used methylcelecoxib (DMC), a close structural analogue of celecoxib lacking COX-2-inhibitory activity. Due to the fact that COX-2 inhibition is the main characteristic of these substances (with exception of methylcelecoxib), we conducted all experiments in COX-2-overexpressing (HCA-7) and COX-2-negative (HCT-116) human colon cancer cells, in order to elucidate whether the observed effects after coxib treatment depend on COX-2 inhibition. Cell survival was assessed using the WST proliferation assay. Apoptosis and cell cycle arrest were determined using flow cytometric and Western blot analysis. The in vitro results were confirmed in vivo using the nude mouse model. Among all coxibs tested, only celecoxib and methylcelecoxib decreased cell survival by induction of cell cycle arrest and apoptosis and reduced the growth of tumor xenografts in nude mice. None of the other coxibs (rofecoxib, etoricoxib, valdecoxib, lumiracoxib) produced anti-proliferative effects, indicating the lack of a class effect and of a role for COX-2. Our data emphasize again the outstanding anti-proliferative activity of celecoxib and its close structural analogue methylcelecoxib in colon carcinoma models in vitro and in vivo.     

Par-4 downregulation confers cisplatin resistance in pancreatic cancer cells via PI3K/Akt pathway-dependent EMT

Cisplatin (CDDP) efficiency in pancreatic cancer therapy is limited due to development of drug resistance. However, the comprehensive mechanisms remain largely unclear. In this study, we first established a CDDP-resistant pancreatic cancer cell line-BXPC-3/CDDP from its parental cell line-BXPC-3. The results showed that CDDP resistance in BXPC-3/CDDP cells correlates with changes in cellular EMT phenotypes. Prostate apoptosis response-4 (Par-4) expression at both mRNA and protein levels were reduced in CDDP-resistant BXPC-3/CDDP cells compared with that in BXPC-3 cells. Ectopic expression of Par-4 reversed EMT and CDDP resistance in BXPC-3/CDDP cells. In BXPC-3 cells, knockdown of Par-4 expression induces EMT and CDDP insensitivity, however, these effects were blocked by inhibition of PI3K/Akt pathway using LY294002. Furthermore, Par-4 knockdown could significantly stimulate PI3K/Akt signaling in BXPC-3 cells. In vivo studies, xenograft BXPC-3 tumors were sensitive to CDDP treatment. Treatment with CDDP alone had little effect on the growth of Par-4 siRNA-transfected BXPC-3 tumors in nude mice and the survival rate compared with control. Inhibition of PI3K/Akt pathway using LY294002 reversed CDDP resistance in Par-4 siRNA-transfected BXPC-3 tumors. In conclusion, these results indicate that Par-4 downregulation confers CDDP resistance via PI3K/Akt pathway-dependent EMT in BXPC-3 cells. Therefore, Par-4 may be a potential target for overcoming CDDP resistance in pancreatic cancer.     

Paclitaxel-octreotide conjugates in tumor growth inhibition of A549 human non-small cell lung cancer xenografted into nude mice

Targeted chemotherapy is a novel approach to cancer therapies. This study evaluated the anti-tumor effects of conjugates made by coupling cytotoxic paclitaxel to the somatostatin analog octreotide in A549 human non-small-cell lung cancer (NSCLC) cells xenografted into nude mice. Two cytotoxic somatostatin analogs, paclitaxel-octreotide and 2paclitaxel-octreotide, were prepared by the coupling of one or two paclitaxel molecules with an octreotide molecule. A549 xenografts expressed mRNAs for type 1, 2, 4, and 5 somatostatin receptors. Immunohistology revealed that type 2 somatostatin receptors were mainly located in tumor cell membrane but type 5 somatostatin receptors were found in tumor cell membrane and cytoplasm. Significant tumor growth inhibition was achieved by 2paclitaxel-octreotide at 150 nM/kg and 300 nM/kg. 2paclitaxel-octreotide also significantly extended the tumor doubling time and significantly reduced tumor microvessel density at these doses. Moreover, there was more fragmented DNA in the 2paclitaxel-octreotide single and double dose groups than in the controls. Paclitaxel was ineffective and more toxic than the conjugate as shown by the significant decline of body weight in Paclitaxel group on Days 6, 12, and 26 compared to those treated with 2paclitaxel-octreotide (P<0.05). White blood cell counts in the paclitaxel single and double dose groups were also significantly less than in the controls (P<0.05). In conclusion, the targeting conjugate 2paclitaxel-octreotide made by coupling two molecules of cytotoxic paclitaxel to one somatostatin analog octreotide could enhance tumor growth inhibition and reduce toxicity in comparison to using the cytotoxic paclitaxel alone.     

番茄红素对荷肝癌裸小鼠移植瘤生长的作用及机制研究

目的研究番茄红素(Lycopene,LP)对荷肝癌裸小鼠移植瘤生长的影响及其机制。方法通过背部皮下注射接种肝癌Hep G2细胞的方法建立荷肝癌裸小鼠模型,并分别腹腔注射5、10、20 mg/(kg·d)的LP和2 mg/(kg·d)的顺铂进行干预治疗,疗程为14 d。治疗后,观察各组裸小鼠的一般生存状态及肿瘤生长状况,称取瘤体质量并计算抑瘤率;通过苏木精-尹红(HE)染色观察肿瘤组织形态结构改变,末端标记法(TUNEL)检测肿瘤组织细胞凋亡状况,免疫组织化学法(IHC)观察肿瘤组织凋亡相关蛋白(caspase-3、Bax、bcl-2)表达。结果经LP治疗14 d,荷肝癌裸小鼠饮食状况和精神状态明显改善,瘤体减小,活动度和硬度降低,肿瘤组织呈现细胞皱缩、片状坏死等病理性改变,凋亡细胞数量明显增多;瘤体质量显著减轻、抑瘤率显著升高;肿瘤组织中caspase-3和Bax表达显著上调,而bcl-2显著下调,Bax/bcl-2比值显著升高;LP上述作用均具有一定的剂量依赖性。结论番茄红素对荷肝癌裸小鼠移植瘤生长具有一定的抑制作用,其机制可能与番茄红素调节凋亡相关基因蛋白表达而促进肿瘤组织细胞凋亡有关。     

血管生成抑制剂联合分化诱导剂治疗结肠癌的实验研究

目的通过建立人LOVO细胞结肠癌裸鼠皮下移植瘤模型,研究血管生成抑制剂（TNP-470）联合分化诱导剂（RA）抗结肠癌生长的效应,二者是否具有协同效果。方法建立人LOVO细胞结肠癌裸鼠皮下移植瘤模型,模型建立后分组治疗,按治疗方式分为对照组、TNP-470组、RA组和联合组（TNP-470＋RA）至治疗结束时处死裸鼠,测量皮下移植瘤重量,计算抑瘤率;检测肿瘤组织结肠癌细胞凋亡指数（AI）。结果裸鼠皮下移植瘤瘤重、抑瘤率,联合组和对照组及TNP-470组、RA组比较差异均有统计学意义（P〈0.05）,且TNP-470组、RA组和对照组之间差异亦有统计学意义（P〈0.05）。治疗组与对照组癌细胞AI比较,差异有统计学意义（P〈0.05）;且联合组与RA组和TNP-470组癌细胞的AI比较,差异也有统计学意义（P〈0.05）。结论血管生成抑制剂TNP-470与分化诱导剂RA联合治疗结肠癌裸鼠皮下移植瘤比TNP-470或RA单药治疗具有更强的抑瘤作用,TNP-470通过抑制肿瘤血管生成,切断肿瘤细胞的血供和营养供给,抑制结肠癌裸鼠皮下移植瘤的生长;而RA通过诱导肿瘤细胞凋亡,降低VEGF的表达抑制肿瘤的生长,二者联和应用,抗肿瘤效应明显增强。     

艾瑞昔布联合5-氟尿嘧啶对裸鼠结肠癌移植瘤侵袭和转移的影响

目的探讨艾瑞昔布联合5-氟尿嘧啶(5-Fu)对结肠癌HT-29细胞裸鼠移植瘤侵袭和转移的影响以及与环氧化酶-2(COX-2)、血管内皮生长因子-C(VEGF-C)、基质金属蛋白酶-9(MMP-9)的关系。方法 40只裸鼠构建人结肠癌HT-29细胞裸鼠皮下移植瘤模型,随机分为4组,每组10只,对照组(0.9%NaCl);艾瑞昔布组[100 mg/(kg·d)];5-Fu组(20 mg/kg);联合用药组[艾瑞昔布100 mg/(kg·d),5-FU 20 mg/kg],艾瑞昔布ig给药,1次/d,5-Fu ip给药,每3天1次,连续给药14 d。实验结束时测量各组裸鼠瘤体大小,并计算抑瘤率;酶联免疫吸附试验(ELISA)检测COX-2、VEGF-C、MMP-9血清浓度,实时荧光定量PCR(qRT-PCR)检测COX-2、VEGF-C、MMP-9的m RNA表达,免疫组化法检测瘤体微血管密度(MVD),Westernblotting法检测COX-2、VEGF-C、MMP-9蛋白表达。结果艾瑞昔布组、5-Fu组、联合用药组与对照组比较,抑瘤效果显著,其中以联合用药组抑瘤效果最佳(P<0.05)。艾瑞昔布组、5-Fu组、联合用药组较对照组裸鼠血清COX-2、VEGF-C、MMP-9浓度、瘤体COX-2、VEGF-C、MMP-9的m RNA和其蛋白表达、瘤体MVD均显著降低(P<0.05),其中以联合用药组降低最显著(P<0.05)。结论艾瑞昔布联合5-Fu可协同抑制结肠癌裸鼠移植瘤侵袭和转移,增强5-Fu的抗肿瘤效果,其作用机制可能与下调COX-2、VEGF-C、MMP-9表达有关。     

胡桃醌体外抗胰腺癌细胞机制的初步研究

目的：探讨胡桃醌体外抗胰腺癌细胞的机制。方法：采用MTT法检测胡桃醌对BXPC-3细胞的生长抑制作用,流式细胞仪分析胡桃醌对BXPC-3细胞周期的影响。结果：MTT法试验结果显示,胡桃醌对体外培养的人胰腺癌BXPC-3细胞具有明显的生长抑制作用,24、48h的IC50值为1.64×10-5、5.8×10-5mol/L。同时胡桃醌能随着浓度的增加,时间的延长,诱导BXPC-3细胞周期停滞于G2期和S期,呈G2期和S期阻滞作用。结论：胡桃醌对胰腺癌BXPC-3细胞有明显的生长抑制作用,并能阻止其周期,诱导BXPC-3细胞凋亡。     

In-vivo effects and mechanisms of celecoxib-reduced growth of cyclooxygenase-2 (COX-2)-expressing versus COX-2-deleted human HCC xenografts in nude mice

We previously reported that celecoxib, a cyclooxygenase-2 (COX-2) inhibitor, suppresses growth of human hepatocellular carcinoma (HCC) cells through both COX-2 dependence and independence. Recently, we established COX-2-deleted human HCC cells, C2D-HuH7, and C2D-HuH7-bearing nude mice. Using this novel model, we examined the pharmacological effects and mechanisms of celecoxib on in-vivo growth of HCC xenografts in relation to COX-2 expression. After treatment with celecoxib, the mice bearing HuH7 or C2D-HuH7 xenografts were assessed for the pharmacological effects and mechanisms of celecoxib on HCC xenograft growth in relation to COX-2 expression. Celecoxib resulted in an effective and comparable growth reduction of both COX-2-expressing and COX-2-deleted HuH7 xenografts in association with decreased Ki-67 expression. These results demonstrated celecoxib's COX-2-independent in-vivo anti-HCC effects. Celecoxib increased peroxisome proliferator-activated receptor gamma predominantly in HuH7 xenografts, indicating its COX-2 dependency. Celecoxib reduced p-Rb and DP1/E2F1 complex predominantly via upregulated p21/CDK4 complex in HuH7 xenograft, but p27/CDK4 complex in C2D-HuH7 xenografts. The effects of celecoxib on phosphatase and tensin homolog deleted on chromosome ten/PI3K/Akt signaling were COX-2 independent, but its effects on extracellular-regulated kinase signaling seemed COX-2 dependent. In addition, the effects of celecoxib on AC-H3, AC-H4, and histone deacetylase 2 could be both COX-2 dependent and independent. In conclusion, celecoxib suppresses growth of HuH7 xenografts regardless of COX-2 expression, which may be mediated through different signaling.     

顺铂联合8-硝基白杨素对人卵巢癌细胞裸鼠移植瘤生长的影响

目的 研究顺铂联合8-硝基白杨素对人卵巢癌COC1细胞裸鼠移植瘤生长的影响。方法 建立人卵巢癌COC1细胞裸鼠皮下移植瘤模型,随机分成4组,每组5只：生理盐水组、8-硝基白杨素组、顺铂组、8-硝基白杨素+顺铂组。观察各组裸鼠移植瘤体积、重量及裸鼠体质量的变化;裸鼠血清乳酸脱氢酶、谷丙转氨酶、肌酐值和外周血白细胞计数的变化;FCM测定瘤组织细胞凋亡率;Western blot分析瘤组织细胞凋亡的可能分子生物学机制。结果 与顺铂组和8-硝基白杨素组相比,顺铂联合8-硝基白杨素联合作用COC1细胞裸鼠移植瘤后,移植瘤的体积、移植瘤的重量均明显降低(P<0.05);与顺铂组相比,顺铂联合8-硝基白杨素联合组裸鼠体质量无明显差异(P>0.05),裸鼠血清乳酸脱氢酶、谷丙转氨酶、肌酐值和外周血白细胞计数无明显差异(P>0.05);顺铂和8-硝基白杨素联合作用COC1细胞16 d后,COC1细胞发生凋亡,同时bcl-2蛋白表达降低,caspase-3蛋白表达增高。结论 顺铂联合8-硝基白杨素通过降低bcl-2蛋白表达,活化caspase-3蛋白表达来抑制人卵巢癌COC1细胞裸鼠移植瘤生长。