人肝癌细胞系MHCC97中趋化因子差异表达研究

目的 检测人肝癌细胞系MHCC97-L和MHCC97-H中趋化因子的mRNA表达水平,了解不同转移潜能的癌细胞系在组成性表达趋化因子方面的异同,寻找影响癌细胞生长、侵袭和转移的关键性趋化因子。方法 收获不加任何刺激的转移性由低到高的人肝癌细胞MHCC97-L和MHCC97-H。AGPC法提取总RNA,以持家基因GAPDH为对照,逆转录-聚合酶链反应(RTPCR)分析C、CC、CXC和CX3C族趋化因子代表的XCL1(Ltn)、CCL5(RANTES)、CXCL8(IL-8)、CX3CL1(Fkn)等的mRNA水平。结果 代表不同家族的4种趋化因子基因在两种细胞系中均呈组成性表达,但XCL1和CCL5在两种细胞系中的表达差异有显著性(P=0.021,P=0.042)。结论 肝癌细胞可组成性地表达目前所知的4个家族的趋化因子,其表达水平可因癌细胞的转移潜能不同而异,提示具有不同结构特征的趋化因子可能以自分泌或旁分泌方式参与癌细胞生长及转移的调节,进一步阐明这些趋化因子产生的意义及其调控机制是有必要的。     

不同来源的CC类趋化因子5对人乳腺癌MCF-7细胞侵袭能力影响及其作用机制研究

目的 观察不同来源的CC类趋化因子5(CC chemokine ligand 5,CCL5)对人乳腺癌细胞侵袭能力影响并探讨其作用机制.方法 CCL5特异性siRNA慢病毒载体转染人乳腺癌MCF-7细胞,分别用RT-PCR和Western Blot检测细胞CCL5 mRNA和蛋白表达水平,并用细胞侵袭实验检测转染前后细胞侵袭能力的变化.同时,以不同浓度的外源性rhCCL5 (recombinant human CCL5) 作为趋化因素,用细胞侵袭实验检测MCF-7细胞侵袭能力及CC类趋化因子受体5(CCR5)单克隆抗体细胞侵袭能力的影响.结果 CCL5-siRNA慢病毒载体感染可有效降低CCL5 在MCF-7细胞内的表达,细胞的侵袭指数无明显改变,干扰组和阴性对照组侵袭指数差异无统计学意义(P>0.05).rhCCL5可以明显诱导MCF-7细胞的侵袭(P<0.05),并且呈浓度依赖趋势;但是这种诱导作用可以部分地被CCR5单克隆抗体阻断,阻断前后细胞的侵袭指数分别为(7.51±0.77)和(3.34±0.51),差异有统计学意义(P<0.05).结论 细胞内表达的CCL5水平变化对乳腺癌细胞MCF-7的侵袭能力没有影响,而外源性CCL5 可以明显增强MCF-7细胞的侵袭,其机制之一可能是通过与细胞表面的CCR5受体结合.     

CCL22-CCR4轴在胃癌腹膜乳斑转移的机制研究

目的 探讨巨噬细胞衍生因子MDC/CCL22-CCR4反应轴在胃癌腹膜乳斑转移中的作用.方法 用RT-PCR和Western blot方法检测CCR4在5种胃癌细胞系中的表达;MTT法测定不同浓度CCL22对胃癌细胞株BGC-823增殖率的变化;Transwell小室体外侵袭实验检测CCL22对胃癌细胞株BGC-823趋化活性的影响;免疫组化检测标本中CCR4表达;单变量分析CCR4表达与胃癌临床病理参数的关系.结果 CCR4mRNA和CCR4蛋白在5种胃癌细胞系中均有表达.CCL22可以促进BGC-823细胞的增殖和趋化.CCR4在胃癌中有表达,CCR4表达水平与胃癌分化程度相关(P<0.05),而与患者的年龄、肿瘤大小、位置、分期以及淋巴结转移等均无关(P>0.05).结论 CCL22-CCR4反应轴可能在胃癌腹膜乳斑转移中起作用.CCR4可能成为预防和治疗胃癌腹膜转移的潜在靶点.     

CCR7对Huh-7肝癌细胞系增殖和侵袭的影响

目的 探讨CCR7对肝细胞肝癌转移的影响。方法 构建siRNA-CCR7载体,稳定转染表达CCR7的肿瘤细胞株Huh-7,用MTT法检测沉默CCR7基因对Huh-7肝癌细胞系增殖的影响,通过趋化侵袭实验检测其对肿瘤细胞株Huh-7趋化、侵袭能力的影响。结果 通过抑制Huh-7细胞CCR7的表达,稳定转染siRNA-CCR7能有效抑制Huh-7细胞的增殖,并能抑制CCL21刺激的趋化和侵袭能力。结论 沉默CCR7基因能有效抑制肝癌细胞增殖和侵袭性。     

miR-542-3p对肝癌细胞恶性生物学行为的影响及机制探讨

目的 探讨microRNA-542-3p(miR-542-3p)在人肝癌细胞系及组织中的表达水平及其对肝癌细胞增殖、侵袭转移能力的影响.方法 采用RT-PCR检测miR-542-3p在肝癌细胞系(HCCLM3、Hep3B、Huh7、SMMC-7721、MHCC-97H、MHCC-97L)以及人正常肝细胞LO2中的表达;...     

长链非编码RNA GAS5在原发性肝癌中的表达及侵袭作用

目的:探讨长链非编码RNA Gas5(lncRNA Gas5)在原发性肝癌组织中的表达及其对Hep3B细胞的侵袭作用。方法:采用RT-PCR检测50例肝癌患者肿瘤和癌旁组织中lncRNA Gas5的表达水平,分析其与临床病理特征的相关性;通过在Hep3B细胞中过表达lncRNA Gas5,利用Transwell小室检测过表达lncRNA Gas5后肿瘤细胞侵袭变化,RT-PCR和Western blot检测PTEN蛋白表达变化。结果:肝癌组织中的lncRNA Gas5表达水平较癌旁组织显著降低(t=9.759,P0.01),并与肝门淋巴结转移(t=7.748,P0.001)和肿瘤TNM分期(t=2.489,P=0.016)相关;过表达lncRNA Gas5后的Hep3B细胞侵袭能力减弱(P0.01),PTEN基因和蛋白表达水平升高(P0.05)。结论:lncRNA Gas5在肝癌组织中的表达下调增强肝癌细胞的侵袭能力,可能成为原发性肝癌术后判断复发的分子标志物。     

质粒介导外源性microRNA下调CCR1降低人肝癌细胞侵袭性

目的观察趋化因子受体CCR1在高转移潜能人肝癌细胞HCCLM3侵袭中的作用。方法设计并合成3对靶向CCR1mRNA的寡核苷酸片段，将其连接pcDNA6．2-GW／EmGFP-miR载体构建表达microRNA（miRNA）真核载体S1、S2、S3，脂质体转染法将重组质粒转入HCCLM3细胞。实时荧光定量聚合酶链式反应（real-time PCR）分析CCR1mRNA抑制效率；Westem印迹测定CCR1蛋白的表达量改变；MTT法检测HCCLM3增殖的影响；Matrigel侵袭实验检测HCCLM3细胞侵袭力的改变。结果转染48h后，S3组CCR1mRNA较空转染组（Mock）和非特异序列组（Neg）减少80％以上，且明显少于其他组（P〈0．01）；72h后S3组CCR1蛋白表达明显少于其他组；转染后，HCCLM3细胞的增殖改变差异无统计学意义。Matrigel侵袭实验结果示转染S3后HCCLM3细胞穿过人工基底膜的数量减少80％，显著低于其他组（P〈0．01）。结论人肝癌细胞HCCLM3的CCR1表达可被质粒导入的外源性miRNA有效抑制，进而降低HCCLM3细胞侵袭性，但对细胞增殖无明显影响。     

SiRNA干扰LI-cadherin对荷人肝癌裸鼠移植瘤的影响

目的 研究肝肠粘连蛋白(liver intestine cadherin,LI-cadherin)在荷瘤裸鼠体内对肝癌细胞Hep3B侵袭转移能力的作用.方法 体外培养肝癌细胞株Hep3B,并进行针对LI-cadherin的SiRNA转染.将Hep3B细胞及转染后的肿瘤细胞接种入裸鼠脾脏,建立裸鼠荷瘤模型.观察成瘤及转移效果,免疫组化检测转移灶组织结构,Western blot方法检测转染前后荷瘤裸鼠体内肿瘤转移灶中LI-cadherin的表达.结果 (1)SiRNA质粒转染入Hep3B肝癌细胞,转染率达到80％.(2)建立裸鼠荷人肝癌模型,30只裸鼠移植手术后均成活,荷瘤率达到60％以上.SiRNA转染组荷瘤率为80％,转移灶个数为26个,明显高于空白对照组和空质粒转染组.(3)裸鼠体内转移灶中Ll-cadherin 的含量SiRNA转染组明显低于空白对照组和空质粒转染组.结论 LI-cadherin对肝癌细胞的黏附能力有重要作用,对LI-cadherin进行SiRNA干扰可增强肝细胞癌在裸鼠体内的侵袭转移能力.     

长链非编码RNA MALAT1在肝癌中表达及功能研究

目的:探讨长片段非编码RNA(lnc RNA)MALAT1在肝癌中的表达及其的功能。方法:用q RT-PCR方法检测80例肝癌患者手术切除的癌组织及其癌旁组织,以及不同肝癌细胞系(Hep G2、Hep3B、HCCLM3、Hu H7)与正常肝细胞系(L02)中MALAT1的表达。分别用MTT试验、划痕试验、流式细胞术检测肝癌细胞(Hep G2、Hu H7)转染MALAT1 si RNA后增殖、迁移、凋亡的变化。结果:80例配对标本中,72例(90%)肝癌组织MALAT1表达较其癌旁组织明显上调,MALAT1在不同肝癌细胞系中的表达水平均不同程度明显高于正常肝癌细胞(均P0.05);Hep G2、Hu H7转染MALAT1 si RNA后,增殖率均呈时间依赖性降低、划痕愈合能力均明显减弱(均P0.05),细胞凋亡率分别为17.0%、16.41%,而各自的对照组分别为8.89%、6.56%,差异有统计学意义(P0.05)。结论:MALAT1在肝癌中的表达上调,且可能与肝癌的恶性生物学行为密切相关,对其作用机制的进一步研究,有望为肝癌的治疗找到新的靶点。     

CCL5及其受体在乳腺癌组织中的表达及其意义

目的检测趋化因子CCL5及其受体在浸润性乳腺癌组织中的表达并探讨其临床意义。方法应用SYBR Green Ⅰ real time-PCR定量检测33例浸润性乳腺癌患者癌组织及其癌旁组织的CCL5及其受体CCR1、CCR3、CCR5 mRNA的表达。结果各期癌组织CCR1及CCR3 mRNA的相对表达量与癌旁正常组织差异无统计学意义（P〉0．05），CCR5及CCL5 mRNA的相对表达量均高于癌旁正常组织（P〈0．05），Ⅰ期和Ⅱ期乳腺癌组织CCL5 mRNA的表达量差异无统计学意义（P〉0．05），Ⅲ期乳腺癌组织CCL5 mRNA的相对表达量为2．477，Ⅳ期乳腺癌组织CCL5 mRNA的相对表达量最高（3．710）。CCR5及CCL5 mRNA的表达高度相关（Pearson相关系数为0．971）。结论趋化因子CCL5及其受体CCR5 mRNA在Ⅳ期乳腺癌组织中的表达水平最高，CCL5可能以自分泌方式与受体CCR5结合促进乳腺癌生长和转移。     

结直肠癌细胞株趋化因子受体的表达及其趋化实验

目的 观察结直肠癌细胞株趋化因子受体CXCR4的表达及其趋化实验,探讨CXCR4在结直肠癌转移中的作用.方法 培养结直肠癌细胞株HT-29、SW480,采用逆转录-聚合酶链反应(RT-PCR)、免疫组织化学染色、Western blot检测结直肠癌细胞株的趋化因子受体CXCR4表达.采用SDF-1诱导的趋化实验模型检测两种结直肠癌细胞株的趋化活动.结果 免疫组织化学检测每100个HT-29或SW480细胞表达CXCR4蛋白的阳性率分别为48.7%和52.6%,RT-PCR和Western blot检测显示CXCR4呈阳性表达;趋化实验表明区化因子SDF-1处理组的细胞移动计数(HT-29:27.6±3.9;SW480:25.3±4.4)显著高于空白对照组(HT-29:9.6±2.5;SW480:9.8±2.1)和抗体处理组(HT-29:9.1±3.1;SW480:9.3±2.0).结论 结直肠癌细胞株可表达CXCR4,趋化因子SDF-1可以趋化结直肠癌细胞.     

The chemokine receptor CXCR4 is more frequently expressed in breast compared to other metastatic adenocarcinomas in effusions

This objective of this study was to investigate the expression of chemokine receptors in tumor cells and leukocytes in breast carcinoma effusions. The expression of leukocyte markers (CD3/4/8/14/16/19) and chemokine receptors (CXCR1/4, CCR2/5/7) was studied in 21 breast carcinoma effusions using flow cytometry. Breast carcinoma cells expressed CXCR4 in 7/21 (33%) effusions, with less frequent expression of CXCR1, CCR5, and CCR7. CXCR2 and CCR2 were absent. Lymphocytes showed frequent CXCR4, CCR5, and CCR7 expression, while CXCR1, CXCR2, CCR2 were rarely or never detected. Macrophages expressed all six receptors except for CXCR2. Comparative analysis of breast carcinoma effusions with previously studied ovarian and cervical/endometrial adenocarcinomas (ACs) showed significantly higher CXCR4 expression in breast carcinoma cells compared to the other gynecological ACs (p = 0.001). Breast and cervical/endometrial carcinoma effusions showed different expression of chemokine receptors in lymphocytes (lower CXCR1, higher CXCR4 and CCR7 levels; p = 0.012, p = 0.005, p < 0.001, respectively) and macrophages (higher CCR7 levels; p < 0.001), as well as lower CD8 counts (p < 0.001) and higher CD19 counts (p = 0.001) compared to ovarian carcinoma effusions. Higher numbers of CD8-positive lymphocytes (p = 0.080) and higher CCR7 monocyte expression (p = 0.087) were associated with a trend for shorter disease-free survival. In conclusion, breast carcinoma cells express CXCR4, a unique feature among metastatic ACs in effusions, with rare expression of other chemokine receptors. Chemokine receptor expression in leukocytes and lymphocyte counts significantly differ from those of ovarian carcinoma effusions. The prognostic role of CCR7 expression in monocytes and CD8 counts in breast carcinoma effusions merits further research.     

趋化因子受体CCR7及CXCR4在结肠癌组织中的表达及意义

目的:观察趋化因子受体CCR7及CXCR4在结肠癌组织中的表达及与临床病理特征的关系,探讨其评价结肠癌侵袭转移及预后的意义.方法:对110例行结肠癌根治术的结肠癌组织标本采用免疫组织化学方法检测CCR7及CXCR4的表达.结果:CCR7、CXCR4在110例结肠癌组织中阳性表达率分别为59.1%和45.5%,其中淋巴结转移组表达率分别为90.8%、69.2%,无淋巴结转移组表达率分别为13.3%、11.1%,差异有统计学意义(P<0.05).CCR7及CXCR4的表达与结肠癌浸润深度、淋巴结转移、肿瘤分期密切相关(P<0.05),与年龄、性别、肿瘤大小、组织学类型及分化程度无关(P>0.05).CCR7判断结肠癌淋巴结转移的敏感性、特异性、阳性预测值及阴性预测值分别为90.8%、86.7%、90.8%、86.7%;CXCR4判断结肠癌淋巴结转移的敏感性、特异性、阳性预测值及阴性预测值分别为69.2%、88.9%、90.O%、66.7%;联合检测CCR7及CXCR4诊断结肠癌淋巴结转移的准确率为88.1%.结论:趋化因子受体CCR7及CXCR4与结肠癌淋巴结转移密切相关,联合检测CCR7及CXCR4对预测结肠癌淋巴结转移情况、评价预后、判定术后治疗方案具有指导价值.     

趋化因子受体CXCR4、CCR7在胃癌细胞中的表达及其与胃癌细胞侵袭能力的关系

目的 观察趋化因子受体CXCR4及CCR7在不同侵袭能力人胃癌细胞株中的差异表达.方法 逆转录-聚合酶链反应(RT-PCR)及Western blot分析CXCR4及CCR7在人胃癌细胞株MGC803、AGS、BGC823、SGC7901的表达;体外侵袭实验测定4株胃癌细胞的侵袭能力.结果 MGC803、AGS、BGC823、SGC7901中CXCR4 mRNA相对表达量分别为:1.2556±0.1384、0.7943±0.0913、0.4749±0.0744、0.2463±0.0344,CCR7 mRNA相对表达量分别为:0.6071±0.1404、0.5355±0.0750、0.2549±0.0522、0.2466±0.0342,CXCR4及CCR7蛋白表达趋势同其相对应的基因表达趋势基本一致.4株胃癌细胞侵袭实验测定的侵袭细胞数分别为:400.0±18.2、310.0±4.0、110.0±13.9、85.0±9.5.CXCR4在不同侵袭能力的胃癌细胞株中存在差异性表达(P<0.05).结论 CXCR4的表达与胃癌细胞侵袭能力成正相关,CCR7的表达与胃癌细胞侵袭能力无明显相关.     

Frequency of T cell expressing Th1 and Th2 associated chemokine receptor in patients with renal allograft dysfunction

Acute rejection of human renal allografts is a frequent, serious posttransplantation complication, occurring in up to 50% of recipients. Leukocyte recruitment is a central feature of acute allograft rejection. Chemokine receptors are expressed on leukocytes in a cell type-specific manner. Recently CCR5+ and CXCR3+ cells have been observed in allograft biopsy specimens of patients undergoing acute cellular rejection (ACR). Herein we investigated the expression of Th1 (CCR5, CXCR3, and CCR2) and Th2 (CCR4, CCR3, and CCR8)-associated chemokine receptors on CD4 and CD8 T-cell populations. We sought to correlate chemokine receptor expression in peripheral blood T-cell subsets with the types of graft dysfunction (biopsy-proven rejections). In the peripheral blood CD4+ and CD8+ T-cell populations of patients with graft dysfunction, we observed a high frequency of Th1-associated chemokine receptors CCR5+ and CCR2+ but not CXCR3.     

FOXP3 shRNA3对肝癌细胞趋化因子及受体CXCL12、CXCL11、CXCR4、CXCR7作用的研究

目的研究FOXP3 shRNA对肝癌细胞株SMMC-7721和MHCC-97H的趋化因子及受体CXCL12、CXCL11、CXCR4、CXCR7的影响。方法设计三种编码FOXP3 shRNA的FOXP3干扰慢病毒,sh-FOXP3-1-pGreenPuro、sh-FOXP3-2-pGreenPuro和sh-FOXP3-pgreenpuro并分别转染SMMC-7721和MHCC-97H细胞。q-PCR检测各组CXCL12、CXCL11、CXCR4、CXCR7 mRNA的表达情况;Western Blot检测各组CXCL12、CXCL11、CXCR4、CXCR7蛋白的表达情况。结果经菌落PCR和测序验证证明三个FOXP3干扰慢病毒载体构建正确;三种干扰序列中sh-FOXP3-1干扰效果最明显,因此后期实验都使用sh-FOXP3-1进行下面的实验。研究显示:(1)与对照组相比,sh-FOXP3-1组的CXCL12、CXCL11、CXCR4、CXCR7 mRNA表达明显下降,差异具有统计学意义。(2)与对照组相比,sh-FOXP3-1组的CXCL12、CXCL11、CXCR4、CXCR7蛋白表达明显下降,差异具有统计学意义。结论干扰FOXP3的表达后,能减少趋化因子CXCL12、CXCL11、CXCR4、CXCR7的表达。     

Chemoattraction of T cells expressing CCR5, CXCR3 and CX3CR1 by proximal tubular epithelial cell chemokines.

BACKGROUND: Chemokines produced by resident renal cells promote the infiltration of leukocyte subsets. We have analysed the chemotactic responses of CD3+ peripheral blood lymphocytes (PBLs) to factors secreted by proximal tubular epithelial cells (PTEC), assessing the role of chemokines and chemokine receptors in this process. METHODS: By FACS we analysed expression of the chemokine receptors CCR5, CXCR3, CX3CR1, CCR2, CXCR1 and CXCR2 on both freshly isolated and activated PBLs. Using Boyden chambers we studied the chemotactic activity of supernatant from resting and cytokine-stimulated (TNF-alpha and IFN-gamma) PTEC towards PBLs. Soluble recombinant chemokines and blocking antibodies were used to study the role of individual chemokine receptors. Chemokine secretion by PTEC was analysed by ELISA. RESULTS: Only a small proportion of freshly isolated cells expressed the chemokine receptors and there was low grade chemotaxis of these cells towards cytokine-stimulated PTEC supernatant compared with unstimulated PTEC supernatant. After activation, 84% of PBLs expressed CCR5, 90% expressed CXCR3 and 19% expressed CX3CR1. There remained low expression levels of CXCR1, CXCR2 and CCR2. Activated PBLs showed strong chemotactic responses to supernatant from cytokine-stimulated PTEC compared with unstimulated PTEC (P<0.001). Chemotaxis of these cells was inhibited by blocking CCR5, CXCR3 and CX3CR1 by 69%, 71% and 29% respectively, with complete inhibition following combined blockade. ELISA showed high levels of the chemokine RANTES/CCL5 (for CCR5) and IP-10/CXCL10 (for CXCR3) in cytokine-stimulated PTEC supernatant. CONCLUSIONS: Chemokines produced by cytokine activated PTEC promote the selective recruitment of activated T cells via the receptors, CCR5, CXCR3 and CX3CR1. These receptors may be amenable to therapeutic manipulation in renal inflammation.     

A novel small-molecule compound targeting CCR5 and CXCR3 prevents acute and chronic allograft rejection

BACKGROUND: Chemokines and chemokine receptors are critical in leukocyte recruitment, activation, and differentiation. Among them, CC chemokine receptor 5 (CCR5) and CXC chemokine receptor 3 (CXCR3) have been reported to play important roles in alloimmune responses and may be potential targets for posttransplant immunosuppression. METHODS: Fully major histocompatibility complex (MHC)-mismatched murine cardiac and islet transplant models were used to test the effect in vivo of a novel, small-molecule compound TAK-779 by targeting CCR5 and CXCR3 in acute allograft rejection. An MHC class II mismatched cardiac transplant model was used to evaluate its efficacy in chronic allograft rejection. Intragraft expression of cytokines, chemokines, and chemokine receptors was measured by quantitative real-time polymerase chain reaction and by histological analysis. RESULTS: Treatment of TAK-779 significantly prolonged allograft survival across the MHC barrier in two distinct transplant models. The treatment downregulated local immune activation as observed by the reduced expression of several chemokines, cytokines, and chemokine receptors. Thereby, the recruitment of CD4, CD8, and CD11c cells into transplanted allografts were inhibited. Furthermore, TAK-779 treatment significantly attenuated the development of chronic vasculopathy, fibrosis, and cellular infiltration. CONCLUSIONS: Antagonism of CCR5 and CXCR3 has a substantial therapeutic effect on inhibiting both acute and chronic allograft rejection. CCR5 and CXCR3 are functional in the process of allograft rejection and may be potential targets in clinical transplantation in the future.     

Deletion of CCR2 but not CCR5 or CXCR3 inhibits aortic aneurysm formation

BACKGROUND: Microscopic analysis of abdominal aortic aneurysms (AAAs) demonstrates an abundance of infiltrating leukocytes. The chemokine receptors CCR2, CCR5, and CXCR3 are associated with pathways implicated previously in aneurysm pathogenesis. We hypothesized that genetic deletions of CCR2, CCR5, and CXCR3 would limit leukocyte infiltration and aneurysm formation in a mouse model of AAA. METHODS: CCR2(-/-), CCR5(-/-), CXCR3(-/-), and control mice of the same genetic background were subject to periaortic application of calcium chloride. Aortic diameters were measured before aneurysm induction and at harvest 6 weeks later. Diameters were compared using the Mann-Whitney test. Aortas were stained with H&E and trichrome for histologic analysis. Aortic MMP-2 and MMP-9 activities were measured using zymography. RESULTS: Aneurysm formation was attenuated in CCR2(-/-) mice with the final mean aortic diameter less than that of the control mice (P < .01). Histology revealed preservation of the lamellar architecture and decreased inflammatory cells. Aortic MMP-2 and MMP-9 levels were decreased in CCR2(-/-) mice. CCR5(-/-) and CXCR3(-/-) mice demonstrated no protection from aneurysm formation, which was corroborated by the tissue histology showing similar inflammatory cell infiltration and elastin degradation. CONCLUSIONS: The CCR2 receptor is involved directly in AAA formation, whereas the CCR5 and CXCR3 receptors are not.     

Chemokine receptor expression in cutaneous T cell and NK/T-cell lymphomas: immunohistochemical staining and in vitro chemotactic assay

Interactions between chemokines and chemokine receptors are involved in migration and invasion of lymphoma cells. We investigated expression profiles of CXCR3 and CCR4 by immunohistochemistry and flow cytometry, and their biologic behaviors by real-time horizontal chemotaxis assay in cutaneous T cell and NK/T-cell lymphomas (TCLs). Tumor cells in mycosis fungoides (MF) constantly expressed CXCR3 at the patch stage, and expressed CCR4 at the tumor stage and in the folliculotropic variant of MF. Neoplastic cells at the plaque stage expressed CXCR3 and/or CCR4. Sezary cells in the dermis and circulation were positive for CCR4. Epidermotropic atypical cells in pagetoid reticulosis expressed CXCR3. CD30 cells exclusively expressed CCR4 in anaplastic large-cell lymphoma, and CXCR3 and/or CCR4 in lymphomatoid papulosis. In CD8TCL and extranodal NK/TCL characterized by extensive epidermotropism, tumor cells were positive for CXCR3. These data demonstrated preferential expression of CXCR3 in epidermotropic tumor cells, and of CCR4 in dermis-based lymphomas. In chemotaxis assays, CCR4 tumor cells in MF and CXCR3 tumor cells in CD8TCL migrated to thymus and activation-regulated chemokine and inducible protein-10, respectively. Therefore, spatial and temporal interactions between chemokine receptors and their ligands seem to dictate recruitment and retention of lymphoma cells in the skin.