THE EFFECTS OF RETINOIC ACID ON EXPRESSION OF C-MYC, C-FOS IN LEUKEMIC PROMYELOCYTES

The expression of c-myc, c-fos of leukemic promyelocytes (HL-60 and acute promyelocytic leukemia cells) from 18 acute promyelocytic leukemia (APL) patients treated with all-trans retinoic acid (RA) in vitro was studied. There was no expression of c-fos in HL-60 cells and APL cells from 17 patients. But in one case, a slight expression of c-fos in leukemic cells was observed, and the alteration of expression level was found during the treatment of the cells with RA in vitro. The expression of c-myc in HL-60 cells induced by RA was altered, decrease in the early, increase in the middle, and decline in the later stage were found. The c-myc expression in leukemic cells of eighteen APL patients was variable. There was c-myc expression in eleven APL cells, but no expression in the others. The APL cells with c-myc expression were treated with RA in vitro to observe the kinetic changes of c-myc RNA level. The results showed that the expression of c-myc was gradually decreased except in few cases. Using in situ hybridization technique for detecting the alteration of c-myc expression in leukemic cells of two APL patients. the high level of c-myc before RA treatment and low level of c-myc expression after obtaining complete remission induced by RA were found. The possibility of different proto-oncogenes implicated differentiation was discussed.     

大鼠肝癌变过程中细胞癌基因的表达

利用RNA-DNA杂交技术观察到:(1)在二乙基亚硝胺(DENA)诱发大鼠肝癌过程中,含有增生结节肝和肝细胞癌肝中c-Ha-ras、c-Ki-ras、N-ras和c-fos癌基因的表达都较正常肝高,而c-myc的表达无明显改变。(2)在两种不同诱发肝癌起始/促癌模型的起始阶段,c-myc的表达显著增高。在2-乙酰氨基芴作用下,N-ras表达的增高非常显著。c-myc和N-ras的表达似有协同作用。在促癌阶段,c-myc和N-ras的表达接近正常。(3)在起始和促癌过程中,c-fos的表达都显著降低,c-Ha-ras和c-Ki-ras表达变化不显著。对这些结果的意义作了初步讨论。     

肝细胞癌和癌旁肝组织中c－myc，N－ras癌基因的表达

应用生物素标记的ｃ－ｍｙｃ，Ｎ－ｒａｓｃＤＮＡ探针对２１例肝细胞癌石蜡切片进行原位杂交研究，结果显示，在２１例肝癌组织中９例呈ｃ－ｍｙｃ阳性（４２．９％），４例为Ｎ－ｒａｓ阳性（１９％），而癌旁肝组织中仅有４例呈ｃ－ｍｙｃ弱阳性，１例呈Ｎ－ｒａｓ弱阳性．Ｃ－ｍｙｃ，Ｎ－ｒａｓ，ｍＲＮＡ多分布于肝癌细胞胞浆中，显微分光光度计检测结果显示，肝癌细胞中ｃ－ｍｙｃ，Ｎ－ｒａｓ，ｍＲＮＡ阳性信号明显高于癌旁肝细胞．作者认为，ｃ－ｍｙｃ，Ｎ－ｒａｓ癌基因在肝细胞癌中均可呈较高水平表达．二者参与了肝癌的恶性转化过程并在肝癌的发生过程中相互协同以维持肝癌的恶性表型．     

急性白血病细胞凝集素受体的研究

用8种生物素标记的凝集素,对44例急性白血病细胞进行亲和细胞化学标记。发现急性淋巴细胞白血病、急性粒细胞白血病与急性单核细胞白血病细胞在花生凝集素受体、荆豆凝集素受体及大豆凝集素受体表达方面存在明显区别,说明不同亚型的急性白血病细胞上的糖基表达是不同的。     

Expression and Fuactional Role of HERG1, K~+ Channels in Leukemic Cells and Leukemic Stem Cells

In order to investigate the expression and functional role of HERG1 K channels in leukemic cells and leukemic stem cells (LSCs), RT-PCR was used to detect the HERG1 K channels expression in leukemic cells and LSCs. The functional role of HERG1 K channels in leukemic cell proliferation was measured by MTT assay, and cell cycle and apoptosis were analyzed by flow cy- tometry. The results showed that herg mRNA was expressed in CD34 /CD38-, CD123 LSCs but not in circulating CD34 cells. Herg mRNA was also up-regulated in leukemia cell lines K562 and HL60 as well as almost all the primary leukemic cells while not in normal peripheral blood mononuclear cells (PBMNCs) and the expression of herg mRNA was not associated with the clinical and cytoge- netic features of leukemia. In addition, leukemic cell proliferation was dramatically inhibited by HERG K channel special inhibitor E-4031. Moreover, E-4031 suppressed the cell growth by induc- ing a specific block at the G1/S transition phase of the cell cycle but had no effect on apoptosis in leukemic cells. The results suggested that HERG1 K channels could regulate leukemic cells prolif- eration and were necessary for leukemic cells to proceed with the cell cycle. HERG1 K channels may also have oncogenic potential and may be a biomarker for diagnosis of leukemia and a novel potential pharmacological target for leukemia therapy.     

细胞癌基因在人脑胚胎发育过程中的表达

作者用分子杂交技术观察了正常4～7月龄胎儿脑的各个部位和正常人脑胚胎发育过程中癌基因的表达。结果显示:多数细胞癌基因的表达水平在小脑和枕叶较高,在颞叶的表达水平较低;人脑组织胚胎发育过程中,c-myc、L-myc、N-myc基因的表达情况类似;erbB基因在胚胎期脑中均有较强的表达,而在正常成人脑中未见其表达;Ha-ras、c-fos基因在人脑胚胎发育的中后期表达水平较高;mos基因在新生儿脑中表达最强。研究结果提示:这7种癌基因可能在人脑的正常胚胎发育过程中起重要作用。     

The expression of oncogenes in the development of human brain]

Seven oncogenes (c-myc, L-myc, N-myc, v-erbB, c-fos, Ha-ras and mos) were used as the probe to detect the total RNA of every part (cerebellum, temporal folium, frontal folium and occipital folium) in human brain from 2 cases of 6 mon fetus and the total RNA in fetal development (4 mon, 5 mon, 6 mon, 7 mon, and newborn) of human brain tissue by RNA dot hybridization analysis. The results showed that most oncogenes were expressed at high level in cerebellum and occipital folium of 6 mon fetus and at low level in temporal folium; there was similar expression of c-myc, L-myc and N-myc in fetal development of human brain tissue, and the highest level was in the 5 mon fetal brain. There was no expression of erbB gene in normal human brain. The high level expression of Ha-ras and c-fos genes was in the middle and later stages of fetal development. These results suggested that c-myc, L-myc, N-myc, erbB, Ha-ras, c-fos and mos genes might play an important role in the fetal development of human brain.     

Proto-oncogenes expression in the process of asthma airway remodeling

Objective: To observe the expression of proto-oncogenes in the process of airway remodeling in asthma. Methods: Guinea pig was used as an asthma model challenged by ovoglobulin. Dot-blot, Northern-blot molecular hybridization and immunohistochemistry techniques were used to detect the expression of c-fos, c-myc, c-jun and c-sis. Results: Expression of c-fos and c-myc mRNA could not be detected or detected at very low level in the control group. There were greatly increased expression of c-fos and c-myc mRNA after guinea pigs were challenged by ovoglobulin. Thirty minutes after the challenge, the expression of c-fos and c-myc mRNA reached to the peak and returned to normal level 4 h after the challenge. Immunohistochemistry studies showed that Fos, Myc, Jun and Sis expressed at low level in control group and increased after ovoglobulin stimulation. Immunohistochemically positive cells laid in the plasma of airway epithelium, in cell nucleus of bronchial epithelium and in the inflammatory cells. Pathologic     

TRAIL基因在白血病原代细胞中的表达

目的：检测白血病原代细胞和正常人外周血淋巴细胞中TRAIL基因的表达情况，探讨TRAIL凋亡途径在肿瘤免疫中的作用。方法：通过RT-PCR检测TRAIL基因的表达。结果：83％（10/12)淋巴系白血病和80％(16／20)髓系白血病患者TRAIL基因表达阳性，正常人外周血淋巴细胞TRAIL基因表达阴性，IL-2能诱导PBL的TRAIL基因表达。结论：大多数白血病原代细胞TRAIL基因表达阳性，可能在肿瘤的自发坏死和凋亡以及免疫逃逸中发挥作用。另外，活化的T细胞表达TRAIL基因，TRAIL可能参与CD95非依赖性激活诱导的细胞凋亡。     

间接免疫酶标法检测白血病细胞表面的运铁蛋白受体

用间接免疫酶标法检测急性单核细胞性白血病(急单)、急性粒细胞性白血病(急粒)、急性淋巴细胞性白血病(急淋)、慢性粒细胞性白血病(慢粒)、慢性淋巴细胞性白血病(慢淋)患者外周血细胞表面的运铁蛋白受体(TR)。47例患者中43例阳性,4例阴性;而48例正常血象者及非白血病患者外周血细胞的TR均为阴性。表明白血病细胞表面TR增加。用此法检测2例白血病及5例正常骨髓片TR阳性细胞的百分率与形态检查幼稚细胞总数的百分率基本相符,而1例急性巨核细胞性白血病及1例急淋骨髓片TR阳性细胞百分率明显高于幼稚细胞,提示白血病细胞TR表达可能较早于形态变异。     

Expression and fuactional role of HERG1, K<Superscript>+</Superscript> channels in leukemic cells and leukemic stem cells

Summary In order to investigate the expression and functional role of HERG1 K⁺ channels in leukemic cells and leukemic stem cells (LSCs), RT-PCR was used to detect the HERG1 K⁻ channels expression in leukemic cells and LSCs. The functional role of HERG1 K⁺ channels in leukemic cell proliferation was measured by MTT assay, and cell cycle and apoptosis were analyzed by flow cytometry. The results showed that herg mRNA was expressed in CD34⁺/CD38⁻, CD123⁻ LSCs but not in circulating CD34⁺ cells. Herg mRNA was also up-regulated in leukemia cell lines K562 and HL60 as well as almost all the primary leukemic cells while not in normal peripheral blood mononuclear cells (PBMNCs) and the expression of herg mRNA was not associated with the clinical and cytogenetic features of leukemia. In addition, leukemic cell proliferation was dramatically inhibited by HERG K⁺ channel special inhibitor E-4031. Moreover, E-4031 suppressed the cell growth by inducing a specific block at the G1/S transition phase of the cell cycle but had no effect on apoptosis in leukemic cells. The results suggested that HERG1 K⁺ channels could regulate leukemic cells proliferation and were necessary for leukemic cells to proceed with the cell cycle. HERG1 K⁺ channels may also have oncogenic potential and may be a biomarker for diagnosis of leukemia and a novel potential pharmacological target for leukemia therapy. LI Huiyu, female, born in 1960, Associate Professor This project was supported by a grant from National Science Foundation for Distinguished Young Scholars of China (No. 30225038).     

艾灸对实验性类风湿关节炎滑膜细胞原癌基因 c-fos和c-myc mRNA表达的影响

目的观察艾灸对实验性类风湿关节炎滑膜细胞原癌基因c-fos、c-myc mRNA表达的影响,初步探讨艾灸对RA滑膜细胞内分子信号传导的调控.方法在日本大耳白兔右后足踝关节部皮内注射福氏完全佐剂造模,经过艾灸治疗,通过半定量RT-PCR测定c-fos和c-myc mRNA的表达量.结果治疗组c-fos和c-myc mRNA表达量显著低于造模组(P＜0.01).结论艾灸治疗能够降低c-fos和c-myc mRNA的表达,影响生长因子信号传导系统.     

Expression of oncoproteins c-fos and c-jun in hypertrophic scars and chronic dermal ulcers and their regulation of basic fibroblast growth factor

目的　探讨c fos与c jun两种原癌基因产物在溃疡与增生性瘢痕创面表达的特征与规律以及与不同组织修复结局发生的关系。方法　 16例标本均取自于外科手术患者 ,其中增生性瘢痕 8例 ,溃疡创面 8例 ,另有正常对照皮肤 5例取自增生性瘢痕患者作为对照。用免疫组化法 (ABC法 )检测c fos与c jun两种癌蛋白以及bFGF在三种组织切片的分布特征。结果　在正常皮肤c fos与c jun的阳性表达主要见于表皮基底细胞和少量皮下成纤维细胞 ,但在相应部位c jun的表达较c fos为弱。在增生性瘢痕 ,c fos与c jun均出现强阳性表达 ,主要见于成纤维细胞。在溃疡组织 ,c fos与c jun的联合表达多见于毛细血管内皮细胞、部分炎症细胞以及成纤维细胞胞浆。结论　增生性瘢痕与溃疡创面c fos与c jun表达量与部位同正常皮肤存在显著差异 ,提示这两种原癌基因产物在影响和调控组织修复中有重要作用。     

白血病细胞系中B-Raf与Raf-1的表达及活性

目的：研究白血病细胞及正常人外周血淋巴细胞中的Ｂ－Ｒａｆ和Ｒａｆ－１表达水平及激酶活性。方法：利用Ｗｅｓｔｅｒｎ印迹法检测Ｂ－Ｒａｆ和Ｒａｆ－１表达水平,利用免疫沉淀及Ｗｅｓｔｅｒｎ印迹法检测Ｒａｆ－１及Ｂ－Ｒａｆ的激酶活性。结果：Ｒａｆ－１在白血病细胞及正常人外周血淋巴细胞中均有表达,表达水平基本相似,其激酶活性较低;Ｂ－Ｒａｆ仅在白血病细胞中表达,且在Ｊｕｒｋａｔ和Ｋ５６２细胞中的表达水平较高,其激酶活性也较高。结论：在白血病细胞中,Ｂ－Ｒａｆ的高表达及活化可能是白血病发病的机制之一。     

三七花总皂苷对内皮素诱导的人主动脉血管平滑肌细胞c-myc、c-fos表达的影响

目的:观察三七花总皂苷(PNFS)对内皮素(ET-1)诱导的人主动脉血管平滑肌细胞(HASMC)癌基因c-myc、c-fos表达的影响。方法:采用ET-1诱导HASMC异常增殖,应用RT-PCR方法检测PNFS对c-myc、c-fos mRNA表达的影响。结果:PNFS能明显抑制HASMC原癌基因c-myc mRNA表达(P<0.05),而对c-fos mRNA表达影响不明显。结论:PNFS可影响癌基因c-myc mRNA表达以抑制HASMC异常增殖。     

人脑原发性肿瘤癌基因转录表达研究

作者以c-myc、L-myc、N-myc、Ha-ras、c-fos、v-erbB基因为探针,对5例正常人脑组织及29例人脑原发性肿瘤组织总RNA进行斑点杂交分析。结果表明:c-erbB和c-fos基因表达增强的百分率最高,分别为88.2％和82.4％;其次是c-myc,为46.1％;L-myc和N-myc仅在少数样品中表达增强,分别为29.3％和31.6％,而Ha-ras基因在正常人脑及脑瘤中表达无显著差异。此外,还观察到在同一脑瘤样品中有2种或2种以上癌基因表达增强现象。上述结果提示c—erbB和c—fos基因在人脑原发性肿瘤发病中起重要作用,以及在人脑瘤发生中可能涉及多个癌基因的变化。     

Survivin基因在急性白血病患者中的表达及其临床意义

目的:检测凋亡抑制基因Survivin在急性白血病(AL)患者骨髓细胞中的表达变化,探讨其与AL的发生、发展及预后的关系。方法:选取40例初发急性白血病患者为实验组和8例正常的骨髓细胞作对照组,采用逆转录PCR(RT-PCR)方法检测两组骨髓细胞SurvivinmRNA的表达情况以及采用免疫组化SP法检测Survivin蛋白的表达情况,分析急性白血病阳性表达和阴性表达间完全缓解率(CR)的差别,采用卡方检验进行统计学分析。结果:40例AL病人骨髓细胞SurvivinmRNA阳性表达率为67.5%,显著高于正常对照组12.5%(P<0.05);Survivin蛋白在急性白血病骨髓细胞中的阳性表达率65.0%(26/40)明显高于正常对照组12.5%(1/8)(P<0.05),骨髓细胞Survivin表达阴性者化疗后骨髓完全缓解率(CR)为70.0%,明显高于阳性表达者的CR为30.0%(P<0.05)。结论:(1)Survivin的过度表达与急性白血病的发生发展有关,其阳性表达可能显示急性白血病的预后不良;(2)Survivin基因表达在白血病细胞的增殖分化和(或)抗凋亡过程中发挥了重要作用,可能是导致AL细胞对化疗药物不敏感的原因之一,Survivin基因可作为靶向白血病治疗的一个潜在靶点。     

肾母细胞瘤基因衍生肽诱导细胞毒性T淋巴细胞对白血病患者骨髓CD34+细胞的体外杀伤效应

目的 探讨肾母细胞瘤基因(WT1)衍生肽负载树突状细胞(DC)诱导细胞毒性T淋巴细胞(CTL)对白血病CD34+细胞的体外清除效应.方法 合成一段针对HLA-A*0201锚位的WT19聚肽,体外负载来源于HLA-A*0201*健康人的DC后,诱导产生WT1肽特异性CTL(A组),以噻唑盐(MTT)比色法观察其对WT1表达阳性白血病患者(HLA-A* 0201+者3例,HLA-A*0201-者3例)骨髓CD34+细胞、健康人(HLA-A*0201+者2例,HLA-A*0201-者1例)外周血CD34+细胞和白血病NB4、K562及U937细胞株的体外杀伤效应,粒细胞-巨噬细胞系集落形成试验观察其对白血病患者骨髓CD34+细胞和健康人外周血CD34+细胞粒细胞-巨噬细胞系集落形成单位(CFU-GM)形成的影响.设立单独DC诱导CTL(B组)和IL-2诱导T细胞(C组)作为对照.结果 在效靶比为20:1时,A组CTL对3例HLA-A*0201+白血病患者骨髓CD34+细胞和NB4细胞的杀伤活性(分别为55.3%±2.8%,67.1%±3.2%、49.4%±3.8%和55.0%±3.7%)明显高于对3例HLA-A*0201-白血病患者骨髓CD34+细胞、健康人外周血CD34+细胞及K562、U937细胞的杀伤活性(均<20%),并明显高于B组和C组CTL(均P＜0.01).2例HLA-A*0201+白血病患者骨髓CD34+细胞经A组CTL处理后CFU-GM集落相对形成率分别为17.8%±4.0%和20.8%±3.4%,明显低于经B组CTL处理后(分别为88.9%±3.4%和91.8%±5.7%,均P＜0.01);HLA-A*0201-白血病患者骨髓CD34+细胞、健康人外周血CD34+细胞经A组和B组CTL处理后CFU-GM集落相对形成率差异尤统计学意义.结论 WT1肽特异性CTL能够以HLA-1类抗原限制方式杀伤高表达WT1基因的白血病CD34+细胞,且能特异性抑制其CFU-GM集落形成,WT1基因的表达产物可以作为清除白血病CD34+细胞靶点.     

外周血单个核细胞内长型和短型瘦素受体的表达

目的探讨肥胖者和正常体重者外周血单个核细胞内长型瘦素受体（OB-RL）和最短的膜结合瘦素受体（OB-Rs）的表达。方法从30例肥胖者（体重指数〉25kg／m^2）和20名正常体重者（体重指数〈23kg／m^2）中获取单个核细胞,利用TRIzoL等分离试剂提取细胞内的总RNA。应用RT-PCR定量研究OB-Rs和OB-RL的表达水平,用放免法检测受试者血清中瘦素的水平。结果在所有受试者中都存在OB-Rs表达;38例受试者存在OB-RL表达,12例肥胖的受试者（BMI〉39kg／m^2）不存在OB-RL mRNA表达。所有受试者的OB-RS表达均高于OB-RL至少4倍以上,无男女差别。肥胖者OB-Rs和OB-RL表达水平比正常体重均显著下降,血清瘦素水平显著升高（6．08ng／ml or 31．21ng／ml）。瘦素受体表达水平与体重指数及血清瘦素水平之间存在显著的负相关。结论外周血单个核细胞内存在OB-Rs和OB-RL的表达,无性别差异,并且OB-Rs具有显著的优势性。