c-kit蛋白在恶性黑素瘤中的表达

目的　探讨c kit蛋白在人恶性黑素瘤中的表达及临床意义 ,分析它和临床病理参数的关系。方法　采用免疫组化S P法检测 44例原发性皮肤恶性黑素瘤、9例转移性恶性黑素瘤及 2 0例良性痣中c kit蛋白的表达。结果　3 4例原发性皮肤恶性黑素瘤、4例转移性恶性黑素瘤、5例良性痣表达c kit蛋白 ,阳性率分别为 77.3 %、44 .4%、2 5 .0 % ,前者的阳性表达率显著高于后两者 (P均 <0 .0 5 ) ;原发性皮肤恶性黑素瘤中浅表扩散性恶性黑素瘤 (SSM )的c kit蛋白阳性表达率显著高于其它类型 (P均 <0 .0 1) ;c kit蛋白的表达与年龄 性别 发病部位 是否淋巴结转移等临床病理因素均无关 (P均 >0 .0 5 )。结论　c kit蛋白的表达可能与人恶性黑素瘤的发生发展有关 ,其有望成为治疗黑素瘤的有效靶向分子之一     

内皮素对黑素瘤A375细胞转化生长因子-β1及磷酸化Smad 3蛋白表达的调节

Objective To investigate the effects ofendothelin-1 (ET-1) and endothelin-3 (ET-3) on the expression of transformation growth factor-beta 1 (TGF-β1) and phosphorylation of Smad 3 in malignant melanoma cell line, A375. Methods Cultured A375 cells were classified into 5 groups, i.e. control group (no stimulation), ET-1 group (stimulated with ET-1), ET-1+BQ123 group (treated with ET-1 and BQ123),ET-1 + BQ788 group (treated with ET-1 and BQ788), ET-3 group (stimulated with ET-3), to receive different stimulation. The working concentrations were 0, 0.1, 1, 10, 100 nmol/L for ET-1 and ET-3, 10 μmol/L for BQ123 and BQ788. After another 12- and 24-hour culture, ELISA, RT-PCR and Western blot were used to detect the expression of TGF-β1 protein and mRNA as well as phosphorylated Smad 3 (P-Smad 3). Results The expression of TGF-β1 in A375 cells was up-regulated by ET-1, but down-regulated by ET-3, and both of the effects were in a concentration-dependent manner. Under the stimulation with ET-1 and ET-3 of 100 nmol/L, the level of TGF-β1 reached 1289.38 ± 89.42 ng/L per 105 cells and 85.09 ± 9.37 ng/L per 105 cells, respectively, significantly different from that in unstimulated cells (both P < 0.05). BQ123 signifi-cantly blocked the up-regnlatory effect of ET-1 on the expression TGF-β1 protein(P < 0.05), but BQ788 had no significant influence on the effect, so was the case with TGF-β1 mRNA. Western blot revealed that ET-1significantly elevated the expression of P-Smad 3 in A375 cells (P <0.05), and the elevation was significantly inhibited by BQ123, but not by BQ788. The expression of P-Smad 3 was statistically decreased by ET-3 in A375 cells (P <0.05). Conclusions The expression of TGF-β1 could be enhanced by ET-1, but suppressed by ET-3. It is likely that endothelin receptor A mediates the phosphorylation of Smad 3 induced by ET-1.     

TAP1-EGFP和TAP2-EGFP融合蛋白在恶性黑素瘤细胞系中的表达

目的 探讨pTAP1-EGFP和(或)pTAP2-EGFP转入恶性黑素瘤细胞株后TAP表达的变化,观察TAP在A375中表达后的业细胞定位.方法 在恶性黑素瘤细胞株A375中转入pTAP1-EGFP和(或)pTAP2-EGFP,G418筛选稳定转染细胞株,检测转染后TAP1和TAP2表达水平的变化.将pDsRed2-ER和pTAP1-EGFP或pTAP2-EGFP共转染入A375细胞内,激光共聚焦显微镜下观察TAP1-EGFP和TAP2-EGFP融合蛋白的亚细胞定位.流式细胞仪检测转染前后细胞表面HIA-Ⅰ的表达.结果 将pTAP1-EGFP和(或)pTAP2-EGFP转染A375细胞株后筛选出稳定克隆.转染pTAP1-EGFP和(或)pTAP2-EGFP后能明显增加A375细胞TAP1和TAP2在蛋白水平的表达,并能增加细胞表面HLA-Ⅰ的表达.共转染pDsRed2-ER和pTAP1-EGFP或pTAP2-EGFP后,在激光共聚焦显微镜下观察,发现TAP1-EGFP和TAP2-EGFP融合蛋白的绿色荧光能够与pDsRed2-ER的红色荧光重叠.结论 构建的pTAP1-EGFP和(或)pTAP2-EGFP质粒在A375细胞系中表达成为TAP1-EGFP和TAP2-EGFP融合蛋白后,能准确定位在内质网上,为研究TAP诱导的后续免疫效应提供基础.     

凋亡抑制蛋白c-FLIP在银屑病患者外周血和皮损中的表达

目的 探讨凋亡抑制蛋白c-FLIP在寻常性银屑病患者外周血和皮损中的表达和分布情况。方法 采用流式细胞仪检测30例寻常性银屑病患者和20例正常人对照组外周血T细胞和B细胞内c-FLIP蛋白表达阳性率，采用免疫组化方法检测c-FLIP蛋白在其皮损中的表达。结果 进行期银屑病患者外周血T细胞内c-FLIP表达（6.32% ± 1.17%）明显高于恢复期患者（2.64% ± 0.74%，P < 0.01）和正常人对照组（2.28% ± 0.54%，P < 0.05）。而其在外周血B细胞内的表达3组间差异无统计学意义，3组分别为0.78% ± 0.16%，0.71% ± 0.32%，0.69% ± 0.18%，P值均 > 0.05。c-FLIP蛋白在进行期银屑病患者皮损中的表达（89.73 ± 5.24）明显高于恢复期（117.40 ± 7.50，P < 0.05）和正常人对照组（121.58 ± 7.93，P < 0.01），恢复期患者和正常人对照组之间差异无统计学意义（P > 0.05）。结论 凋亡抑制蛋白c-FLIP在进行期银屑病患者的外周血T细胞和皮损内明显高表达，可能参与银屑病患者T细胞的增殖。     

Janus激酶抑制剂在斑秃治疗中的研究进展

斑秃(AA)是一种常见的自身免疫性非瘢痕性秃发,病因复杂,部分患者治疗效果不佳或反复发作。随着对疾病机制的认识逐渐深入,新的治疗靶点和药物被发现。其中,已有研究报道AA患者使用Janus激酶(JAK)抑制剂后出现毛发再生,JAK-STAT信号通路已成为AA临床治疗干预的目标。本文将对JAK-STAT信号通路在AA中的作用以及JAK抑制剂治疗AA的研究进展进行综述。     

E-钙黏素、P-钙黏素在寻常型银屑病皮损中的表达及其意义

目的:探讨E-钙黏素和P-钙黏素在寻常型银屑病皮损中的表达及其意义。方法:采用免疫组化SP法检测21例寻常型银屑病皮损和10例正常表皮组织中E-钙黏素、P-钙黏素的表达情况。结果:①P-钙黏素在正常表皮组织仅表达于基底细胞层;在银屑病皮损中广泛表达于表皮全层,表达强度较正常表皮组织也明显增加(P<0.001);②E-钙黏素在银屑病皮损及正常表皮组织中均是表皮全层表达;表达强度也未见明显改变(P>0.05)。结论:P-钙黏素可能与银屑病的发病有关。     

内皮素对黑素瘤A375细胞转化生长因子-β1及磷酸化Smad 3蛋白表达的调节

Objective To investigate the effects ofendothelin-1 (ET-1) and endothelin-3 (ET-3) on the expression of transformation growth factor-beta 1 (TGF-β1) and phosphorylation of Smad 3 in malignant melanoma cell line, A375. Methods Cultured A375 cells were classified into 5 groups, i.e. control group (no stimulation), ET-1 group (stimulated with ET-1), ET-1+BQ123 group (treated with ET-1 and BQ123),ET-1 + BQ788 group (treated with ET-1 and BQ788), ET-3 group (stimulated with ET-3), to receive different stimulation. The working concentrations were 0, 0.1, 1, 10, 100 nmol/L for ET-1 and ET-3, 10 μmol/L for BQ123 and BQ788. After another 12- and 24-hour culture, ELISA, RT-PCR and Western blot were used to detect the expression of TGF-β1 protein and mRNA as well as phosphorylated Smad 3 (P-Smad 3). Results The expression of TGF-β1 in A375 cells was up-regulated by ET-1, but down-regulated by ET-3, and both of the effects were in a concentration-dependent manner. Under the stimulation with ET-1 and ET-3 of 100 nmol/L, the level of TGF-β1 reached 1289.38 ± 89.42 ng/L per 105 cells and 85.09 ± 9.37 ng/L per 105 cells, respectively, significantly different from that in unstimulated cells (both P < 0.05). BQ123 signifi-cantly blocked the up-regnlatory effect of ET-1 on the expression TGF-β1 protein(P < 0.05), but BQ788 had no significant influence on the effect, so was the case with TGF-β1 mRNA. Western blot revealed that ET-1significantly elevated the expression of P-Smad 3 in A375 cells (P <0.05), and the elevation was significantly inhibited by BQ123, but not by BQ788. The expression of P-Smad 3 was statistically decreased by ET-3 in A375 cells (P <0.05). Conclusions The expression of TGF-β1 could be enhanced by ET-1, but suppressed by ET-3. It is likely that endothelin receptor A mediates the phosphorylation of Smad 3 induced by ET-1.     

凋亡抑制蛋白c-FLIP在带状疱疹患者细胞免疫中作用的研究

目的探讨凋亡抑制蛋白c-FLIP在抗水痘-带状疱疹病毒(VZV)免疫中的作用。方法采用流式细胞术检测30例无恶性肿瘤的带状疱疹患者、17例伴恶性肿瘤的带状疱疹患者和20例正常对照者外周血T细胞和B细胞内c-FLIP蛋白表达阳性率和CD3+T细胞阳性率。结果急性期伴有恶性肿瘤的带状疱疹患者外周血T细胞内c-FLIP表达明显低于无恶性肿瘤和正常对照组(P<0.05,P<0.01),无恶性肿瘤组明显低于正常对照组(P<0.05);恢复期无恶性肿瘤的带状疱疹患者外周血T细胞内c-FLIP表达明显增加,与急性期相比有明显差异(P<0.01),但恢复期的伴恶性肿瘤和急性期相比无明显差异(P>0.05);外周血B细胞内c-FLIP表达在急性期和恢复期的带状疱疹患者中无明显差异(P>0.05)。CD3+T细胞在带状疱疹患者外周血中的阳性率与c-FLIP有相似的表达趋势,两者呈正相关(r分别为0.806和0.534,P均<0.05)。结论凋亡抑制蛋白c-FLIP在急性期带状疱疹患者外周血T细胞内低表达,恢复期则高表达,且与T细胞数量明显正相关,表明其可能参与带状疱疹患者T细胞的增殖,在带状疱疹发生、发展中发挥一定的作用。     

表皮囊肿并发毛母质瘤

报告2例表皮囊肿并发毛母质瘤。例1，男，34岁。右侧大腿上方有一灰褐色肿块，直径4cm。皮损组织病理检查示表皮囊肿的囊壁不完整，囊肿中央和周边均为成片的影细胞，囊肿中央的影细胞内可见许多小而圆的嗜伊红角化中心，提示向毛母质分化，未见嗜碱性细胞。例2，女，20岁。右上臂暗蓝色、质硬的皮下结节，直径1cm。皮损组织病理检查示表皮囊肿与毛母质瘤同时存在于真皮内，相互独立，毛母质瘤内可见嗜碱性细胞和影细胞。2例患者均未出现加德纳综合征。     

内皮素对黑素瘤A375细胞转化生长因子-β1及磷酸化Smad 3蛋白表达的调节

Objective To investigate the effects ofendothelin-1 (ET-1) and endothelin-3 (ET-3) on the expression of transformation growth factor-beta 1 (TGF-β1) and phosphorylation of Smad 3 in malignant melanoma cell line, A375. Methods Cultured A375 cells were classified into 5 groups, i.e. control group (no stimulation), ET-1 group (stimulated with ET-1), ET-1+BQ123 group (treated with ET-1 and BQ123),ET-1 + BQ788 group (treated with ET-1 and BQ788), ET-3 group (stimulated with ET-3), to receive different stimulation. The working concentrations were 0, 0.1, 1, 10, 100 nmol/L for ET-1 and ET-3, 10 μmol/L for BQ123 and BQ788. After another 12- and 24-hour culture, ELISA, RT-PCR and Western blot were used to detect the expression of TGF-β1 protein and mRNA as well as phosphorylated Smad 3 (P-Smad 3). Results The expression of TGF-β1 in A375 cells was up-regulated by ET-1, but down-regulated by ET-3, and both of the effects were in a concentration-dependent manner. Under the stimulation with ET-1 and ET-3 of 100 nmol/L, the level of TGF-β1 reached 1289.38 ± 89.42 ng/L per 105 cells and 85.09 ± 9.37 ng/L per 105 cells, respectively, significantly different from that in unstimulated cells (both P < 0.05). BQ123 signifi-cantly blocked the up-regnlatory effect of ET-1 on the expression TGF-β1 protein(P < 0.05), but BQ788 had no significant influence on the effect, so was the case with TGF-β1 mRNA. Western blot revealed that ET-1significantly elevated the expression of P-Smad 3 in A375 cells (P <0.05), and the elevation was significantly inhibited by BQ123, but not by BQ788. The expression of P-Smad 3 was statistically decreased by ET-3 in A375 cells (P <0.05). Conclusions The expression of TGF-β1 could be enhanced by ET-1, but suppressed by ET-3. It is likely that endothelin receptor A mediates the phosphorylation of Smad 3 induced by ET-1.