C－myc反义寡聚脱氧核苷酸抑制人脑胶质瘤细胞系BT_（325）Myc蛋白合成和细胞增殖并诱导其分化的研究

目的：探讨在体外ｃ－ｍｙｃ反义核酸是否可通过阻断人脑胶质瘤细胞中ｃ－ｍｙｃ基因的表达而抑制细胞增殖并诱导分化．方法：人工合成与ｃ－ｍｙｃｍＲＮＡ起始码及其后四个密码子互补的寡聚脱氧核苷酸（简称反义核酸）片段，用它处理培养的ＢＴ３２５细胞，观察它对细胞增殖的影响．同时用免疫细胞化学方法检测细胞中Ｍｙｃ蛋白的水平以及能反映胶质瘤细胞分化的Ｓ－１００和ＧＦＡＰ两种蛋白的水平，分析这些指标的变化．结果：发现４ｕｍｏｌ／Ｌ的ｃ－ｍｙｃ反义核酸明显抑制ＢＴ３２５细胞的增殖和Ｍｙｃ蛋白的合成，且后者发生在加入反义核酸后１ｈ，并持续２４ｈ以上，而细胞增殖受抑制要到第５日才明显．从第２日到第５日细胞中Ｓ－１００和ＧＦＡＰ染色明显加深，反映细胞有分化趋势．用同样长度的无关序列寡聚脱氧核苷酸作对照，则未见上述变化，表明ｃ－ｍｙｃ反义核酸的作用是序列特异性的．结论：ｃ－ｍｙｃ反义核酸可特异地抑制ＢＴ３２５细胞中Ｍｙｃ蛋白的合成和细胞增殖，并能诱导其分化．     

Neuronal expression of Raf protooncogene in the brain stem of adult guinea pig

The Raf protooncogenes encode for cytoplasmic serine/threonine-specific protein kinases which can be activated via growth factor receptors by phosphorylation. Immunohistochemical and Western blotting studies have proven the existence of Raf protein kinases in neurons of the cerebral cortex of rats and guinea pigs. The aim of the present study was to map the immunohistochemical distribution of Raf kinase-like staining in the brain stem of guinea pig. Polyclonal antibodies were used that were raised against a recombinant viral protein in combination with the avidin-biotin-peroxidase system for detection of immunoreactivity. Specificity of the antibodies was tested in Western blotting experiments. Cytoplasmic immunostaining was observed in motor nuclei of hypoglossal, accessory, vagus, facial, trigeminal, abducent, oculomotor and trochlear nerves, and in the nucleus ambiguus, nucleus retroambigualis, lateral vestibular nucleus, mesencephalic nucleus of the trigeminal nerve, the red nucleus, raphe nuclei and reticular formation. Scattered neurons were stained in other sensory nuclei, such as solitary tract nuclei, medial, dorsal and ventral vestibular nuclei and cochlear nuclei. The spinal trigeminal nucleus and the main sensory nucleus of the trigeminal nerve contained few medium-sized immunoreactive cells. In general, staining was mainly somatodendritic; the axonal plexus was not positive. It is concluded, that the widespread neuronal appearance of cytoplasmic Raf kinase suggests an important role in transmission of trophic and growth factor signals in these neurons.     

表皮生长因子和甲硫氨酸脑啡肽诱导人白细胞c—fos基因表达的研究

由健康人外周血分离淋巴细胞及溶血处理后的全白细胞，以外源性表皮生长因子和甲硫氨酸脑啡肽培育后取细胞液帛晟滴片和滴膜，用生物素标记的ｃ－ｆｏｓｃＤＮＡ探针进行原位杂交和斑点印迹杂交。结果显示表皮生升因子和甲硫氨酸脑啡肽两组ｃ－ｆｏｓ原癌基因的表达对照组增。     

自发性脑卒中诱导原癌基因c－fos在大鼠脑内的表达

用免疫组织化学（ＡＢＣ）方法，对易卒中型肾血管性高血压大鼠高血压晚期自发形成脑卒中时以及高血压早期和中期脑内ｃ－ｆｏｓ原癌基因蛋白（ＦＯＳ）的表达进行了观察．在高血压晚期，假手术组不出现脑卒中，但个别大鼠的扣带皮质和梨状皮质出现微弱的ＦＯＳ表达；手术组都出现脑卒中，其中３例脑内未见ＦＯＳ表达，４例则于大脑皮质、海马、黑质和下丘脑等处出现程度不等的ＦＯＳ表达．高血压早期，手术组和假手术组的下丘脑、隔核、中脑中央灰质和丘脑室旁核出现分布状态和强弱都相似的ＦＯＳ表达．高血压中期，手术组和假手术组脑内未见ＦＯＳ表达．本研究结果提示：（１）高血压晚期，自发性脑卒中能诱导ｃ－ｆｏｓ在脑内的广泛表达，脑内不同部位ＦＯＳ表达的机制和功能意义不同，本文对其进行了讨论；（２）腹部手术能诱导下丘脑等与内脏功能调节有关的功能区ＦＯＳ的表达；（３）高血压早期和中期，通过ｃ－ｆｏｓ表达的方法未能发现高血压对大鼠脑功能活动的影响。     

原癌基因bc1-2在眼科领域的研究进展

原癌基因bc1 2属抗凋亡基因 ,它不促进细胞增殖 ,但能延长细胞的生命期限。Bc1 2在许多细胞增殖性疾病的发生中起着非常重要的作用。通过影响组织细胞内bc1 2基因的表达 ,为临床上一些退行性疾病及细胞增殖性疾病的治疗提供新的思路。Bc1 2在眼科领域的研究主要集中于遗传性和环境因素所致的视网膜变性。Bc1 2的过表达可以阻止实验性缺血、视神经挫伤、光诱导损伤以及一些视蛋白和cGMP磷酸二酯酶缺陷所致的视网膜光感受器凋亡 ,但对由于某些基因突变所致的凋亡无阻止作用     

碘缺乏病区硒与胎儿脑海马区原癌基因表达

目的　研究微量元素硒 (Se)与胎儿脑海马区神经元原癌基因c-fos蛋白表达之间的关系。方法　在陕西省缺碘 (I)低硒区 (宜君县 )缺碘高硒区 (紫阳县 )及对照区 (临潼县 )共采集 5 1例脐血进行微量元素Se、I的检测 ,并在以上 3地共解剖孕 2 0～ 37周的水囊引产胎儿胎脑 2 0例 ,通过免疫组化学法检测胎脑海马区神经元原癌基因c -fos蛋白产物的表达。结果　 3地I水平无显著性差异 ,Se水平有显著性差异 (P <0 0 5 ) ,以紫阳为最高 ,宜君为最低 ;紫阳c-fos蛋白表达较强 ,与宜君相比有显著性差异 (P <0 0 5 )。结论　Se的水平可能影响胎脑海马区神经元c -fos蛋白的表达 ,并进而影响胎儿神经系统及智能的发育     

Multivariate survival analysis of clinicopathologic features in surgical stage I endometrioid carcinoma including analysis of HER-2/neu expression

OBJECTIVE: We previously described vascular invasion-associated changes, defined as the presence of vascular invasion or perivascular lymphocytic infiltrates, as key prognostic indicators in stage I endometrioid carcinoma. The current study was undertaken to examine the prognostic value of HER-2/neu expression in relation to other factors, including vascular invasion-associated changes, in surgical stage I endometrioid carcinoma.STUDY DESIGN: Seventy-one patients with surgical stage I endometrioid carcinoma treated by hysterectomy and followed up were randomly chosen for retrospective analysis of prognostic indicators including standard clinicopathologic features, deoxyribonucleic acid ploidy, and HER-2/neu expression. The latter was examined by an objective computerized quantitative immunohistochemical system.RESULTS: By univariate analysis many factors were found to correlate with outcome, including age, tumor grade, depth of invasion, ploidy, HER-2/neu expression, and vascular invasion-associated changes. By multivariate analysis only vascular invasion-associated changes, aneuploidy, and HER-2/neu overexpression were found to independently correlate with survival. Stratification of patients on the basis of these three features revealed survival rates of 100%, 92%, and 60% when none, one, and two or three features were present, respectively.CONCLUSION: This study suggests that HER-2/neu expression correlated with outcome independent of other factors in endometrial carcinoma and may aid in estimating prognosis. The prognostic value of HER-2/neu overexpression independent of vascular invasion suggests that this factor may operate by increasing the ability of tumor cells to grow at a distal site once vascular invasion occurs.     

紫杉醇对血管平滑肌细胞原癌基因转录和表达的影响

目的观察紫杉醇（paclitaxel）对血管平滑肌细胞原癌基因c-jun、c-fos转录和表达的影响及其量效、时效关系。方法提取大鼠主动脉原代平滑肌细胞，将第4-9代细胞用于实验。分别运用逆转录聚合酶链反应和免疫组织化学技术检测血管紧张素及不同浓度（O．1、1．0和10．0tanol／L）紫杉醇对平滑肌细胞c-iun、c-fos mRNA转录及其蛋白质产物合成的影响。结果高、中、低剂量紫杉醇对血管紧张素诱导原癌基因的转录和表达没有明显作用。结论血管紧张素通过促进c-fos、c-jun的转录和表达引起血管平滑肌细胞的增殖。紫杉醇抗血管平滑肌细胞增殖作用并非通过抑制原癌基因c-fos、c-jun转录和表达来实现。     

Loss of heterozygosity of chromosome 12p does not correlate with KRAS mutation in non-small cell lung cancer

Activating mutations of RAS gene families have been found in a variety of human malignancies, including lung cancer, suggesting their dominant role in tumorigenesis. However, several studies have shown a frequent loss of the wild-type KRAS allele in the tumors of murine models and an inhibition of oncogenic phenotype in tumor cell lines by transfection of wild-type RAS, indicating that wild-type RAS may have oncosuppressive properties. To determine whether loss of wild-type KRAS is involved in the development of human lung cancer, we investigated the mutations of KRAS, NRAS and BRAF in 154 primary non-small cell lung cancers (NSCLCs) as well as 10 NSCLC cell lines that have been shown to have KRAS mutations. We also determined the loss of heterozygosity status of KRAS alleles in these tumors. We detected point mutations of KRAS in 11 (7%) of 154 NSCLCs, with 10 cases at codon 12 and 1 at codon 61, but no mutations of NRAS or BRAF were found. Using the laser capture microdissection technique, we confirmed that 9 of the 11 tumors and 7 of the 10 NSCLC cell lines retained the wild-type KRAS allele. Among the cell lines with heterozygosity of mutant and wild-type KRAS, all of the cell lines tested for expression were shown to express more mutated KRAS than wild-type mRNA, with higher amounts of KRAS protein also being expressed compared to the cell lines with a loss of wild-type KRAS allele. In addition, among 148 specimens available for immunohistochemical analysis, 113 (76%) showed positive staining of KRAS, indicating that the vast majority of NSCLCs continue to express wild-type KRAS. Our findings indicate that the wild-type KRAS allele is occasionally lost in human lung cancer, and that the oncogenic activation of mutant KRAS is more frequently associated with an overexpression of the mutant allele than with a loss of the wild-type allele in human NSCLC development.