CXCR4抑制剂AMD3100对结肠癌肝转移的实验研究

目的:研究CXCR4特异性抑制剂AMD3100治疗结肠癌肝转移的效果.方法:建立结肠癌肝转移动物模型.将18只模型裸鼠随机分为对照组(0.1 mL生理盐水)、低剂量治疗组(AMD3100,4 mg/kg)和高剂量治疗组(AMD3100,6 mg/kg),每组6只,治疗6周后评价疗效.结果:对照组、低剂量治疗组和高剂量治疗组脾脏原发瘤直径大小分别为(13.83±7.73)、(5.17±0.75)和(2.83±1.33) mm(治疗组与对照组、治疗组之间差异均有统计学意义,P<0.05).对照组、低剂量治疗组和高剂量治疗组肝脏转移瘤直径大小分别为 (3.42±2.24)、(2.26±1.12)和(1.62±0.88) mm(治疗组与对照组、治疗组之间差异均有统计学意义,P<0.05).对照组、低剂量治疗组和高剂量治疗组肝脏表面转移结节计数分别为21.00±3.90、13.00±1.79和2.17±1.37 (治疗组与对照组、治疗组之间差异均有统计学意义,P<0.05).结论:CXCR4特异性抑制剂AMD3100可以降低模型动物的结肠癌肝转移程度,抑制肝脏转移瘤的生长,且呈浓度依赖性;SDF-1/CXCR4轴在结肠癌肝脏转移中具有重要作用.     

SDF-1/CXCR4对结直肠癌肝转移瘤表达乙酰肝素酶的影响

目的研究基质细胞衍生因子1（stromal cell derived factor-1,SDF-1）对高表达趋化因子受体CXCR4的结直肠癌Lovo细胞表达乙酰肝素酶（heparanase, HPA）的影响,并探讨 SDF-1/CXCR4轴对裸鼠结直肠癌肝转移瘤模型中乙酰肝素酶表达的影响。方法分别用SDF-1和SDF-1/CXCR4的拮抗剂AMD3100处理直肠癌 Lovo细胞,运用RT-PCR和免疫细胞化学技术检测结直肠癌细胞HPA 的表达;建立裸鼠结直肠癌肝转移瘤模型,检测AMD3100干预和非干预组转移瘤细胞中HPA 蛋白的表达。结果（1）结直肠癌Lovo细胞经不同浓度（50 ng/ml、100 ng/ml、200 ng/ml ）SDF-1处理后,HPA mRNA的表达量逐渐增高。用相同浓度SDF-1分别处理直肠癌Lovo细胞12h和24h后,其HPA的表达量随作用时间的延长而增高。在相同作用时间内,其HPA的表达量随SDF-1作用浓度（50 ng/ml、100 ng/ml 、200 ng/ml）的升高而增高。（2）SDF-1处理组HPA 的mRNA和蛋白表达水平均高于SDF-1+AMD3100处理组和对照组。（3）肝脏转移瘤中HPA的表达量随AMD3100治疗剂量的增高而降低。结论（1）SDF-1可刺激结直肠癌Lovo细胞HPA表达增加,并且这一效应与SDF-1的浓度和作用时间有关。（2）AMD3100能有效抑制SDF-1刺激结直肠癌细胞表达HPA,使得肿瘤细胞侵袭能力下降,从而起到抑制肿瘤转移的作用。     

ELAM-1在鼻咽癌裸鼠移植瘤中的表达及其与转移的关系

目的 建立裸鼠鼻咽癌转移模型并探讨 E-选择素（ELAM-1）与鼻咽癌转移的相关性。方法 将鼻咽癌5-8F细胞悬液注射于裸鼠左后肢爪垫，观察裸鼠状态、成瘤情况并测量裸鼠体重及移植瘤长短径；采用连续病理切片苏木精-伊红染色观察移植瘤及转移情况，将16只人鼻咽癌荷瘤裸鼠分为转移组和非转移组；采用免疫组织化学法检测两组移植瘤组织中ELAM-1的表达。 结果 16只裸鼠均成瘤，成瘤率为100.0%，其中10只裸鼠出现转移瘤，转移率为62.5%。建模前，两组裸鼠体重差异无统计学意义［（13.83±0.56）g vs （14.62±0.30） g，t=1.026，P=0.071］。建模后4~7周，裸鼠瘤体体积呈指数增长，且转移组移植瘤增长速度较非转移组快，非转移组裸鼠瘤体体积小于转移组［（198.91 ± 163.29） mm³ vs （268.76 ±174.31） mm³，t=4.376，P=0.005］。ELAM-1在鼻咽癌裸鼠移植瘤、淋巴结转移灶及远处转移灶中的表达均为阳性，主要表达于细胞膜。转移组移植瘤光密度值高于非转移组（0.4497±0.0705 vs 0.0435±0.0082，t=4.388，P＝0.001）。结论 本研究成功构建稳定性好、转移率高的鼻咽癌裸鼠移植瘤转移模型，且ELAM-1在裸鼠移植瘤中高表达，可促进鼻咽癌裸鼠移植瘤生长和转移。     

脂联素对高脂饮食诱导的胰岛素抵抗子宫内膜癌裸鼠移植瘤生长的影响

目的 探讨脂联素对高脂饮食诱导裸鼠胰岛素抵抗子宫内膜癌移植瘤生长的影响。方法 40只裸鼠随机分成高脂组和普食组,每组20只,分别喂养高脂饲料（high-fat diet,HFD）和普通饲料（normal diet,ND）,10周后测定裸鼠空腹血糖（fasting blood glucose,FBG）和空腹血胰岛素（fasting serum insulin,FINS）水平,计算胰岛素抵抗指数（HOMA-IR）,建立胰岛素抵抗裸鼠模型。第11周接种子宫内膜癌HEC-1B细胞,待瘤体长至0.5 cm时,两组随机选取10只裸鼠腹腔注射脂联素（adiponectin,APN）分为HFD+APN组、ND+APN组;其余裸鼠分别注射0.9%氯化钠分为HFD组、ND组;14周后,测定各组血糖和血脂的代谢情况。结果 高脂组裸鼠10周后的平均体重、体长、FBG、FINS、HOMA-IR均大于普食组,糖耐量试验和胰岛素耐量试验结果显示血糖水平亦高于普食组（均P＜0.05）;接种子宫内膜癌HEC-1B细胞后均成瘤,建模成功。HFD+APN组和ND+APN组移植瘤平均重量和体积增长速度均分别低于HFD组和ND组（均P<0.05）。HFD组和HFD+APN组14周后的FINS、HOMA-IR、TC和TG均较ND组和ND+APN明显升高（均P<0.01）,且HFD组明显高于HFD+APN组（均P<0.01）;HFD+APN组的血脂联素水平低于ND+APN组（P<0.01）,HFD组则低于ND组（P<0.05）。结论 脂联素可改善高脂饮食导致的胰岛素抵抗,抑制子宫内膜癌移植瘤生长。     

抑制CXCR4活性对乳腺癌骨转移影响的体内外研究

目的:应用特异性抑制剂AMD3100抑制人乳腺癌骨高转移MDA-MB-231SA-rfp细胞中CXCR4的活性,探讨CX-CR4在乳腺癌细胞体内、外增殖和迁移中的作用和机制。方法:CCK8法和Transwell法检测AMD3100对MDA-MB-231SA-rfp细胞体外增殖和迁移能力的影响。构建MDA-MB-231SA-rfp细胞骨转移裸鼠模型,以不同质量浓度的AMD3100处理后,X线影像观察骨转移情况,进一步利用MicroPET进行半定量分析,并应用H-E染色检测骨转移灶的定位。Western blotting法检测AMD3100对MDA-MB-231SA-rfp细胞和移植瘤转移灶组织中CXCR4蛋白表达的影响。结果:AMD3100能明显抑制MDA-MB-231SA-rfp细胞在SDF-1刺激下的增殖和迁移(P<0.05),较高质量浓度(2 000 ng/ml)的AMD3100效果更明显(P<0.01)。成功构建MDA-MB-231SA-rfp细胞裸鼠乳腺癌转移模型,不同质量浓度AMD3100处理后,小鼠下肢骨骨质破坏程度降低;MicroPET分析发现,对照组、低剂量AMD3100组、高剂量AMD3100组SUVmax值分别为9.44±0.53、5.70±0.25、2.18±0.47(P<0.01);组织病理检测证实为乳腺癌骨转移灶。Western blotting结果显示,AMD3100作用前后MDA-MB-231SA-rfp细胞和骨转移灶标本中CXCR4蛋白表达无明显变化。结论:AMD3100降低CXCR4的活性能抑制乳腺癌MDA-MB-231SA-rfp细胞体外增殖和迁移能力,并能抑制裸鼠体内乳腺癌骨转移灶的形成。     

蛋白酶体抑制剂硼替佐米对食管鳞状细胞癌生长的影响

目的 探讨蛋白酶体抑制剂硼替佐米对食管鳞状细胞癌生长的影响。方法 基于全基因组测序数据,利用药物靶标数据库DGIdb筛选食管鳞状细胞癌相关药物靶基因,采用DAVID软件进行KEGG信号通路富集分析。采用MTT法检测硼替佐米对KYSE30、KYSE180、KYSE150、TE1、KYSE510等5株食管鳞状细胞癌细胞生长的影响,以DMSO作用为相应对照组。裸鼠体外成瘤后分别腹腔注射生理盐水（对照组）及硼替佐米（硼替佐米组）,观察硼替佐米对裸鼠移植瘤体积及重量的影响;免疫组织化学法检测裸鼠移植瘤细胞Ki-67蛋白的表达。结果 469例食管鳞状细胞癌的基因组学测序数据在DGIdb数据库中共鉴定出307个药物靶基因,突变频率>2.5%的显著突变药物靶基因包括PIK3CA、NOTCH1、CDKN2A、ERBB4等;药物靶基因富集的信号通路有RTK-RAS、PI3K/AKT/mTOR、NOTCH、ERBB信号通路、细胞周期和蛋白酶体途径等。MTT实验结果显示,硼替佐米处理后,5株食管鳞状细胞癌细胞的增殖能力均较对照组降低（P<0.05）;与对照组相比,硼替佐米组裸鼠移植瘤体积减小［（1 909.18±533.40） mm³ vs （1 065.83±283.94） mm³,P＝0.007］,移植瘤重量降低［（1.60±0.36） g vs （0.98±0.30） g,P＝0.009］。免疫组织化学法检测结果显示,与对照组比较,硼替佐米组裸鼠移植瘤组织中Ki-67表达降低（86.32±4.51 vs 43.83±3.22,P＝0.001）。结论 蛋白酶体抑制剂硼替佐米在体外和体内均可抑制食管鳞状细胞癌细胞生长。     

积雪草酸对人肝癌SMMC-7721细胞增殖和自噬的影响

目的  探讨积雪草酸（asiatic acid,AA）对人肝癌SMMC-7721细胞增殖和自噬的影响。方法 不同浓度AA作用于人肝癌SMMC-7721细胞24 h后,MTT法检测细胞活性并观察自噬抑制剂3-MA对40 μmol/L AA的干预作用;MDC染色检测自噬泡的形成,Western blot检测自噬相关蛋白LC3-Ⅰ、LC3-Ⅱ、p62、mTOR、p-mTOR及p53的表达。结果 MTT检测结果显示,不同AA浓度均可抑制人肝癌SMMC-7721细胞增殖,并呈浓度依赖性（F＝46.790,P＝0.006）,IC₅₀ =37.313 μmol/L。与单独使用40 μmol/L AA相比,自噬抑制剂3-MA可部分逆转40 μmol/L AA对SMMC-7721细胞增殖的抑制作用［（46.400±9.099）% vs （22.000±3.391）%,P＜0.001］。MDC染色实验表明40 μmol/L AA干预可增加自噬泡形成。Western blot检测发现,与对照组比较,40 μmol/L AA可明显降低LC3-Ⅰ的蛋白表达,而提高LC3-Ⅱ表达（1.744±0.108 vs 1.529±0.065,t=2.928,P=0.043;0.113±0.031 vs 0.380±0.036,t=-9.754,P<0.001）,降低p62蛋白表达（0.522±0.024 vs 0.123±0.019,t=22.565,P<0.001）和p-mTOR蛋白表达（1.252±0.039 vs 0.353±0.028,t=30.775,P<0.001）,但对mTOR和p53的蛋白表达无影响（1.713±0.111 vs 1.556±0.076,t=1.555,P=0.190;0.671±0.040 vs 0.726±0.055,t=-1.555,P=0.210）。结论 AA能抑制人肝癌SMMC-7721细胞增殖,可能与其通过非p53依赖方式负调控mTOR通路诱发自噬有关。     

miR-125b-5p对喉鳞状细胞癌细胞能量代谢及增殖的影响

目的  探讨miR-125b-5p对喉鳞状细胞癌（laryngeal squamous cell carcinoma,LSCC）细胞能量代谢和增殖的影响及其可能的作用机制。方法 实验分为miR-125b-5p转染组（miR-125b-5p模拟物转染LSCC）和对照组（空质粒转染LSCC细胞）。采用RT-qPCR 检测miR-125b-5p在正常支气管上皮细胞和LSCC细胞中的表达,CCK-8和流式细胞术检测LSCC细胞增殖、凋亡情况,Western blot检测miR-125b-5p过表达后LSCC中HK2的表达,3H-2DG法和乳酸盐比色测定实验分别检测LSCC细胞葡萄糖消耗和乳酸产生的情况。结果 RT-qPCR实验结果显示,与正常支气管上皮细胞相比,LSCC细胞中miR-125b-5p的表达降低（0.68±0.03 vs 0.22±0.05,t=7.025,P=0.001）。CCK-8实验结果显示,转染72 h和96 h后,miR-125b-5p转染组LSCC细胞的增殖能力均较对照组明显降低（P<0.05）。流式细胞术检测结果显示,miR-125b-5p转染组LSCC细胞凋亡率较对照组升高 ［（37.52±2.34）% vs （12.46±3.52）%,t=7.025,P<0.001）］,但细胞集落形成能力降低（0.29±0.02 vs 1.02±0.03,t=5.689,P=0.005）;荧光素酶报告基因测定实验结果显示,miR-125b-5p转染组的荧光素酶活性低于对照组（0.32±0.03 vs 1.01±0.02,t=7.543,P=0.001）;Western blot法实验结果显示,miR-125b-5p转染组HK2蛋白表达水平较对照组降低（0.12±0.02 vs 0.75±0.03,t=5.875,P=0.023）;miR-125b-5p转染组葡萄糖消耗量［（3.85±0.86） dpm/mg vs （10.52±1.34） dpm/mg,t=6.118,P=0.005］以及乳酸产生量［（4.23±1.36） dpm/mg vs （10.96±2.45） dpm/mg,t=5.907,P=0.002］亦较对照组降低。结论 miR-125b-5p可能通过下调HK2表达降低喉鳞状细胞癌细胞的能量代谢和增殖能力,诱导细胞凋亡。     

人类免疫缺陷病毒阳性和阴性弥漫大Ｂ细胞淋巴瘤的临床特点及近期疗效分析

目的 分析比较人类免疫缺陷病毒（HIV）阳性与阴性弥漫大B细胞淋巴瘤患者的临床特点及疗效。方法 收集博茨瓦纳弗朗西斯敦市仰加奎医院肿瘤内科2012年3月至2015年3月诊治的弥漫大Ｂ细胞淋巴瘤患者共71例,其中HIV阳性37例,HIV阴性34例,给予CHOP方案一线化疗,对两组的临床特点及疗效进行分析。结果 与HIV阴性组相比,HIV阳性组B症状发生率高（56.8% vs. 29.4%; P=0.020）,更容易出现胃肠道（37.8% vs. 14.7%; P=0.028）、肝（29.7% vs. 9.7%; P=0.027）、肺（27.0% vs. 9.7%; P=0.048）浸润。HIV阳性组与HIV阴性组治疗完全缓解率分别为18.9%（7/37）与41.2%（14/34）（P=0.040）;客观有效率分别为48.6%（18/37）与70.6%（24/34）（P=0.060）。HIV阳性组化疗后出现贫血、白细胞下降及继发感染比例高于HIV阴性组（均P<0.05）。HIV阳性组中有24例在确诊淋巴瘤前已给予高效价抗逆转录病毒治疗（highly active antiretroviral therapy,HAART）治疗,13例确诊后给予HARRT治疗,其客观有效率分别为41.7%和61.5%（P=0.248）。CD4⁺细胞数>200/mm³和≤200/mm³患者,其客观有效率分别为71.4%和34.8%（P=0.031）。结论 HIV阳性患者就诊时表现出更强的侵袭性。结合HARRT治疗,CHOP方案可使HIV阳性患者达到类似于HIV阴性患者的客观有效率,但完全缓解率低。HIV阳性组患者HARRT起始治疗时间不影响近期疗效。CD4⁺细胞数低是近期疗效不良的预测因素。     

lncRNA SH3PXD2A-AS1在结肠癌组织中的表达及其临床意义

目的 探讨长链非编码RNA（lncRNA） SH3PXD2A-AS1在结肠癌组织中的表达及其临床意义。方法 收集于广西医科大学附属肿瘤医院手术切除的40例结肠癌患者的癌组织及相应癌旁正常组织。采用RT-PCR法检测lncRNA SH3PXD2A-AS1的表达水平。下载TCGA数据库中的结肠癌数据，验证lncRNA SH3PXD2A-AS表达水平与结肠癌临床病理特征及预后的关系。结果 lncRNA SH3PXD2A-AS1在结肠癌组织中的表达水平高于癌旁正常组织（6.53±1.62 vs 4.37±0.96，t=3.445，P=0.002），其在M1期患者中的表达水平高于M0期，在Ⅲ~Ⅳ期患者中患者中的表达水平亦高于Ⅰ~Ⅱ期 （P<0.05），但与患者总生存期无关（χ²=0.326，P=0.586）。TCGA数据库验证结果显示，lncRNA SH3PXD2A-AS1在结肠癌组织中呈高表达，但与患者的总生存期无关（P>0.05）。结论 lncRNA SH3PXD2A-AS1在结肠癌组织中呈高表达，且与肿瘤远处转移有关，可能是判断肿瘤进展程度的指标。     

CXCL12受体抑制剂调控三阴性乳腺癌放射治疗的敏感性

目的 探讨趋化因子12（CXCL12）受体CXCR4抑制剂AMD3100对人乳腺癌MDA-MB 231细胞裸鼠移植瘤的放射增敏效应及其作用机制。方法 建立人乳腺癌裸鼠移植瘤模型,并随机分为4组：对照组、AMD3100处理组、放射治疗组和联合治疗组（AMD3100+放疗）;称量肿瘤的重量并测量移植瘤的体积,计算放射增敏比,绘制肿瘤生长曲线;实时荧光定量PCR（QPCR）检测CXCR4和表皮生长因子受体（EGFR）基因表达;蛋白质印迹法检测CXCR4、EGFR和基质金属蛋白酶-9（MMP-9）蛋白表达。结果 经统计CXCR4抑制剂AMD3100的放射增敏比为1:45。QPCR结果显示,与对照组比较,CXCR4和EGFR基因的相对表达量在AMD3100处理组、放射治疗组及联合治疗组分别下调60％、45％、82％和56％、48％、73％,差异有统计学意义（P＜0.05）。单纯AMD3100治疗或者放射治疗均能使CXCR4、EGFR表达下调（P＜0.05）,联合治疗较单纯AMD3100治疗和放疗更能显著地抑制CXCR4和EGFR的表达（P＜0.05）。Western blotting结果显示,与对照组比较,CXCR4、EGFR及MMP-9在AMD3100处理组、放射治疗组及联合治疗组中的蛋白相对表达量均下调（P＜0.05）。AMD3100与放疗均可抑制CXCR4、EGFR及MMP-9的表达,两者联用较单一治疗的效果更加显著（P＜0.05）。     

Blockade of CXCR4 in oral squamous cell carcinoma inhibits lymph node metastases

We have previously demonstrated that a stromal cell-derived factor-1 (SDF-1; CXCL12)/CXCR4 system is involved in the establishment of lymph node metastasis in oral squamous cell carcinoma (OSCC). In this study, we investigated whether the blockade of CXCR4 inhibits lymph node metastasis in B88 OSCC cells. These cells harbour a functional CXCR4 and have the potential to metastasise to the lymph node in vivo. Following introduction of a vector that expresses short hairpin small interfering RNA (shRNA) against CXCR4, we isolated three clones (shCXCR4-16, -17 and -21) that showed decreased expression of CXCR4 mRNA. These clones also had reduced CXCR4 protein levels and showed impairments in calcium flux and cell migration in response to SDF-1. These cells were orthotopically inoculated into the masseter muscle of nude mice. Lymph node metastases, loss in body weight and tumour volumes were significantly inhibited in mice inoculated with shCXCR4-17 cells compared to mice inoculated with control cells. SDF-1-induced migration of B88 cells was significantly inhibited in vitro by the treatment with 1,1′-[1,4-phenylenebis(methylene)]bis-1,4,8,11-tetraazacyclotetradecane octahydrochloride (AMD3100), a CXCR4 antagonist. Subcutaneous administration of AMD3100 significantly inhibited the lymph node metastases of B88 cells when they were orthotopically inoculated into the masseter muscle of nude mice. Moreover, the enhanced production of interleukin (IL)-6 and IL-8 in response to SDF-1 was inhibited by shRNA against CXCR4 or by treatment with AMD3100. These results suggest that blockade of CXCR4 may be a potent anti-metastatic therapy against lymph node metastases in cases of CXCR4-related OSCC.     

Establishment of a CXCR4-expressing gastric cancer cell line in nude mice and the effect of AMD 3100 on tumor regression

BACKGROUND AND OBJECTIVE: CXCR4 plays a central role in cell migration in metastasis and dissemination of cancer. We have recently established a CXCR4-expressing gastric cancer cell line. Then we have prepared a therapeutic model for it, and assessed its usefulness in the in vivo system. METHODS: Tumors from gastric cancer patients were transplanted into BALB/C (nu/nu) nude mice. After several subcultures in vivo, the tumor cell line was successfully established. In addition, this cell line expressed the CXCR4 receptor. The nude mice bearing these cell line tumors were administered AMD 3100, an inhibitor of the CXCR4 receptor, and its anti-tumor activity was assessed. RESULTS AND DISCUSSION: The tumor volume was reduced by 27.6% in mice administered AMD 3100 compared with the control group (p < 0.02). None of the nude mice showed toxic signs of toxicity. Establishment of a CXCR4 expressing human gastric cancer cell line transplantable into nude mice seems to be important in discussing the possibility of an in vivo model of treatment with AMD 3100 in the in vivo system.     

Involvement of SDF-1alpha/CXCR4 axis in the enhanced peritoneal metastasis of epithelial ovarian carcinoma

Epithelial ovarian carcinoma (EOC) spreads by implantation of tumor cells onto the human peritoneal mesothelial cells (HPMCs) lining the peritoneal cavity. The aim of this study was to determine whether the stromal cell-derived factor-1alpha (SDF-1alpha)/CXCR4 axis is involved in the interaction of EOC cells with HPMCs in peritoneal metastasis. Clinically, we first evaluated CXCR4 expression in sections from 36 primary EOCs using immunohistochemistry. We next examined whether SDF-1alpha played roles in EOC progression, including in proliferation, cell motility, attachment to HPMCs, and the in vivo development of peritoneal metastasis through CXCR4. Of the 36 carcinomas, 16 cases (44.4%) were positive for CXCR4 immunoexpression. Positive CXCR4 expression significantly predicted poorer overall survival compared with negative expression (p = 0.0069). We found CXCR4 expression in both EOC cells and HPMCs. In contrast, the level of production of SDF-1alpha by HPMCs was higher than that by various EOC cells. Functionally, SDF-1alpha induced enhanced attachment between ES-2 cells and HPMCs or extracellular matrix components. The enhancement of adhesion potential by SDF-1alpha was inhibited by AMD3100, a CXCR4 antagonist, and by phosphatidylinositol 3 kinase and p44/42 inhibitors. Furthermore, intraperitoneal treatment with AMD3100 resulted in reduced dissemination in nude mice inoculated with ES-2 cells. The present results suggest that there may be a link between the SDF-1alpha/CXCR4 axis and enhanced intraperitoneal dissemination of EOC and that CXCR4 may be a novel target for the treatment of EOC.     

Biological role and potential therapeutic targeting of the chemokine receptor CXCR4 in undifferentiated thyroid cancer

Anaplastic thyroid carcinoma (ATC) is a rare thyroid cancer type with an extremely poor prognosis. Despite appropriate treatment, which includes surgery, radiotherapy, and chemotherapy, this cancer is invariably fatal. CXCR4 is the receptor for the stromal cell-derived factor-1 (SDF-1)/CXCL12 chemokine and it is expressed in a variety of solid tumors, including papillary thyroid carcinoma. Here, we show that ATC cell lines overexpress CXCR4, both at the level of mRNA and protein. Furthermore, we found that CXCR4 was overexpressed in ATC clinical samples, with respect to normal thyroid tissues by real-time PCR and immunohistochemistry. Treatment of ATC cells with SDF-1 induced proliferation and increase in phosphorylation of extracellular signal-regulated kinases and protein kinase B/AKT. These effects were blocked by the specific CXCR4 antagonist AMD3100 and by CXCR4 RNA interference. Moreover, AMD3100 effectively reduced tumor growth in nude mice inoculated with different ATC cells. Thus, we suggest that CXCR4 targeting is a novel potential strategy in the treatment of human ATC.     

CAFs对乳腺癌细胞生物学特性的影响及机制探讨

目的：通过乳腺癌间质成纤维细胞（ carcinoma-associated fibroblasts,CAFs）与乳腺癌细胞系MDA-MB-231共培养的体外细胞实验及裸鼠接种的在体动物实验,观察CAFs对乳腺癌细胞增殖、凋亡、侵袭和转移活性的影响,并探讨其作用机制。方法：体外实验：分离培养浸润性导管癌组织中CAFs和正常成纤维细胞（ normal fibroblasts,NFs）,然后分别与乳腺癌细胞MDA-MB-231体外共培养,采用MTT法、流式细胞仪、Ma-trigel人工模拟基底膜法分别检测乳腺癌细胞的增殖、凋亡、细胞黏附和侵袭能力。动物在体实验：选择乳腺癌细胞系MDA-MB-231、CAFs和NFs,结合生理盐水（NS）、基质细胞衍生因子-1（SDF-1）及其配体拮抗剂AMD3100,组成不同的组别并接种于裸鼠（共6组）。观察肿瘤的大小,有无淋巴结、肺、肝脏转移。留取血标本及肿瘤组织行SDF-1表达水平的检测。结果：MDA-MB-231与CAFs和NFs共培养后乳腺癌细胞的增殖活性显著增强,其中CAFs的作用较NFs更强（P＝0.011）;CAFs组的黏附能力（34.70±4.84个/视野）明显强于NFs组（20.16±3.09个/视野）,P＝0.000;而CAFs组的侵袭性（89.0±4.62个/视野）也明显强于NFs组（81.6±6.08个/视野,P＝0.045）。CAFs组中的MDA-MB-231的早期凋亡率（2.9±2.4）较NFs组（5.0±4.2）明显降低（P＝0.026）;MDA231﹢CAFs ﹢NS 组的种植肿瘤平均体积最大（9.092±2.662cm3, P＝0.000）;此外,该组共有4只（66.6％）,MDA231﹢NS组有2只（33.3％）存在腋窝淋巴结转移,未见肝肺转移灶。在MDA231﹢CAFs﹢NS组中,血标本 SDF -1值（75.25±16.23pg/ml）、肿瘤组织标本中 SDF -1mRNA值（11.686±8.926）、组织中SDF-1蛋白表达水平（1.006±0.327）均为最高,与其他各组相比均有统计学差异（ P＝0.000）。结论：CAFs可影响乳腺癌肿瘤细胞的生物学特性,具有促进肿瘤细胞增殖,增强其黏附、侵袭及转移能力。其机制可能是通过乳腺癌间质成纤维细胞分泌SDF-1与其特定的受体CXCR4结合这一信号通路来实现的。     

趋化因子受体4和间质细胞衍生因子1α与鼻咽癌器官特异性转移的相关性研究

目的 研究趋化因子受体4(CXCR4)在鼻咽癌细胞中的表达,间质细胞衍牛因子1α(SDF-1α)在鼻咽癌远处靶器官中的表达,探讨CXCR4和(或)SDF-1α在鼻咽癌器官特异性转移中的作用.方法 应用逆转录聚合酶链反应(RT-PCR)和免疫组织化学法分析30例鼻咽癌、15例正常鼻咽组织中CXCR4 mRNA和蛋白的表达及其同临床病理学因素之间的相关性,应用免疫组织化学法分析鼻咽癌患者的正常颈部淋巴结(包括颈深上和颈深下淋巴结)、骨髓、肺、肝脏和肾脏、结肠(各5例)中SDF-1α蛋白的表达.结果 RT-PCR检测结果显示,鼻咽癌组织中CXCR4 mRNA相对表达强度(0.71±0.22)显著高于正常鼻咽组织(0.14±0.07;F=27.94,P<0.05);免疫组织化学检测结果显示,鼻咽癌组织中CXCR4蛋白的表达(1.58±0.59)显著高于正常鼻咽组织(0.51±0.22;F=17.75,P<0.05).鼻咽癌组织中CXCR4 mRNA和蛋白的表达与临床分期、淋巴结转移、细胞分化程度显著相关(均P<0.05).SDF-1α蛋白在鼻咽癌患者的颈深上淋巴结、骨髓、肺、肝脏中表达较高(2.35±0.67),而在颈深下淋巴结、肾脏和结肠中表达较弱(0.68±0.23),差异有统计学意义(t=10.13,P<0.01).结论 CXCR4的表达与鼻咽癌的转移密切相关,CXCR4和(或)SDF-1α在鼻咽癌器官特异性转移中可能发挥重要作用.     

CXCL12-CXCR4 Promotes Proliferation and Invasion of Pancreatic Cancer Cells

Objective: CXCL12 exerts a wide variety of chemotactic effects on cells. Evidence indicates that CXCL12,in conjunction with its receptor, CXCR4, promotes invasion and metastasis of tumor cells. Our objective was toexplore whether the CXCL12-CXCR4 biological axis might influence biological behavior of pancreatic cancercells. Methods: Miapaca-2 human pancreatic cancer cells were cultured under three different conditions:normal medium (control), medium + recombinant CXCL12 (CXCL12 group), or medium + CXCR4-inhibitorAMD3100 (AMD3100 group). RT-PCR was applied to detect mRNA expression levels of CXCL12, CXCR4, matrixmetalloproteinase 2 (MMP-2), MMP-9, and human urokinase plasminogen activator (uPA). Additionally, cellproliferation and invasion were performed using CCK-8 colorimetry and transwell invasion assays, respectively.Results: CXCL12 was not expressed in Miapaca-2 cells, but CXCR4 was detected, indicating that these cells arecapable of receiving signals from CXCL12. Expression of extracellular matrix-degrading enzymes MMP-2, MMP-9, and uPA was upregulated in cells exposed to exogenous CXCL12 (P<0.05). Additionally, both proliferationand invasion of pancreatic cancer cells were enhanced in the presence of exogenous CXCL12, but AMD3100intervention effectively inhibited these processes (P<0.05). Conclusions: The CXCL12-CXCR4 biological axisplays an important role in promoting proliferation and invasion of pancreatic cancer cells.