外周血miR-150-5p、SOCS1 mRNA对类风湿关节炎“病”“证”诊断意义的初步探讨＊

目的 探讨类风湿关节炎患者外周血miR-150-5p、细胞因子信号抑制因子1（suppressor of cytokine signaling 1，SOCS1）mRNA的表达及对类风湿关节炎（Rheumatoid Arthritis，RA）疾病诊断、中医证型判断的意义。方法 纳入符合诊断标准的RA患者57例及健康对照组19例，根据《22个专业95个病种中医诊疗方案》有关RA的中医证候诊断标准，判断RA的中医证型。qPCR检测RA患者及健康对照组miR-150-5p、SOCS1mRNA的相对表达水平，同时检测血常规、肝功能、肾功能等常规指标。双荧光素酶分析方法判断两者是否存在靶向关系。统计分析miR-150-5p、SOCS1 mRNA对RA疾病的诊断意义及其与中医证型的相关性。结果 RA患者外周血miR-150-5p的相对表达水平下调，低于正常人群（t = -19.019，P < 0.05）；其表达水平随疾病活动度升高，有下降趋势；患者外周血SOCS1 mRNA的相对表达水平上调，低于正常人群（t = 5.333，P < 0.05）；其表达水平随疾病活动度升高，有上升趋势。MiR-150-5p与SOCS1 mRNA有靶向结合关系（P < 0.05）。通过AUC曲线比较，miR-150-5p的相对表达水平区分RA的敏感性及特异性分别为98.1%、92.1%（AUC = 0.972，P < 0.05）；SOCS1 mRNA的相对表达水平无法区分RA（AUC = 0.472，P > 0.05）。RA患者中miR-150-5p的相对表达水平低于3.06，RA患者风湿痹阻证、寒湿痹阻证的相对风险分别为8.33、250.00（P < 0.05）。结论 miR-150-5p、SOCS1 mRNA在RA患者中有差异性表达，且有靶向结合关系。miR-150-5p可能是RA的疾病诊断及中医风湿痹阻证、寒湿痹阻证证型诊断的潜在生物标志物。     

奥沙利铂通过调控miRNA-7-5p/RAF-1促进胃癌SGC-7901细胞凋亡的研究

目的探讨奥沙利铂如何调控MAPK通路,抑制胃癌细胞的增殖。方法NCBI检索文献,利用TargetScan、StarBase和miRBase数据库,进行GO分析与KEGG通路富集,找到相关miRNAs,预测靶基因。应用Real-time PCR、MTT、Hoechst33258、流式细胞术、细胞划痕实验、Western blot等方法分析人胃癌SGC-7901细胞的增殖、细胞周期、侵袭及蛋白表达情况。结果胃癌细胞中miR-7-5p显著低表达,RAF1与miR-7-5p存在互靶关系。miR-7-5p mimics与奥沙利铂均可促进SGC-7901细胞的凋亡,提高G1期细胞百分率(P<0.05),降低侵袭、迁移速度。caspase3、caspase9蛋白表达升高,Bcl-2/Bax比值降低(P<0.05)。结论过表达miR-7-5p与奥沙利铂均可促进胃癌SGC-7901细胞的凋亡,提示奥沙利铂可能通过上调miRNA-7-5p促进SGC-7901细胞的凋亡,降低侵袭、迁移速度。     

Immunohistochemical and molecular analysis of PI3K/AKT/mTOR pathway in esophageal carcinoma

Among the numerous signaling pathways involved in tumorigenesis, PI3K‐AKT‐mTOR is a key one that regulates diverse cellular functions. However, its prognostic value in esophageal carcinoma remains unclear. In our study, we examined the immunohistochemical expression of phosphorylated (p‐) AKT, mTOR, p70S6K and 4E‐BP1 along with the mutational status of PIK3CA and AKT1 genes by High Resolution Melting Analysis and Pyrosequencing in 44 esophageal carcinomas. The results were correlated with the clinicopathological characteristics of the patients in an effort to define their possible prognostic significance. Total p‐mTOR cytoplasmic expression, assessed in 10 random areas, was positively correlated with tumor stage (Kruskal–Wallis ANOVA, I/II vs III/IV, p = 0.0500). Μoreover, maximum p‐mTOR cytoplasmic immunoexpression, estimated in hot spot areas, was positively associated with tumor grade (Mann–Whitney U test, I/II vs III, p = 0.0565). Interestingly, p‐4E‐BP1 immunoreactivity was negatively correlated with tumor histological grade (Mann–Whitney U test, I/II vs III, p = 0.0427). No mutation was observed in exons 9 and 20 of PIK3CA gene and in exon 4 of AKT1 gene. In conclusion, our findings depict the presence of activated PI3K/AKT/mTOR pathway in esophageal cancer bringing forward p‐mTOR and p‐4E‐BP1 for their potential role in esophageal carcinogenesis. Additional studies are warranted to validate our findings.     

红藻氨酸致痫后大鼠海马ERK、P38 MAPK和JNK的活性变化

目的研究红藻氨酸(KA)诱导大鼠癫痫发作后海马组织细胞外调节蛋白激酶(ERK)、p38MAPK和c-jun氨基末端激酶(JNK)的活性(磷酸化状态)的变化情况。方法立体定向大鼠侧脑室内注射KA引起大鼠癫痫发作,采用Western-blot方法观察KA致痫后大鼠海马中活性ERK、p38MAPK和JNK的变化。结果KA诱导大鼠癫痫发作后,海马组织ERK、p38MAPK和JNK的磷酸化水平开始增高,分别于30min、1h和30min后达高峰,呈对照组的4.76倍、2.16倍和3.95倍,两组比较差异具有显著性(P<0.01),之后逐渐下降。结论KA致痫大鼠癫痫发作后,海马组织MAPKs的活性产生变化,其信号通路可能参与癫痫发作后海马组织的病理生理反应过程。     

A trip through the signaling pathways of melanoma

Many cellular signaling pathways are involved in the development of cancer. Depending on the tumor entity, the nature as well as the mode of activation can differ. Some signaling pathways frequently show changes as all tumor cells have to fulfill some basic requirements such as independence from growth factors or insensitivity against apoptosis. In this review, the possibilities of a tumor to manipulate signaling pathways to reach these goals are exemplified based on an archetypical melanoma cell. In addition, new therapeutic options based on the knowledge of signaling pathways will be discussed.     

Skp2、p27kip1在大肠癌中的表达及其临床意义

目的研究Skp2、p27kip1在大肠癌组织中的表达,探讨它们之间的关系及其临床意义。方法采用免疫组化方法检测Skp2、p27kip1蛋白在83例大肠癌、18例大肠腺瘤和20例大肠正常黏膜中的表达。结果大肠癌组织中Skp2蛋白阳性率(28.65±12.60)%,显著高于大肠腺瘤组织(17.28±10.66)%(P<0.01)和大肠正常黏膜组织(6.60±5.54)%(P<0.01)。Skp2蛋白表达与细胞分化程度、肿瘤分期及淋巴结转移呈正相关(P<0.01)。大肠癌组织中p27kip1蛋白阳性率(24.61±11.27)%,显著低于大肠腺瘤组织(49.94±13.22)%(P<0.01)和大肠正常黏膜组织(65.40±15.74)%(P<0.01)。p27kip1蛋白表达与细胞分化程度、肿瘤分期及淋巴结转移呈负相关(P<0.01)。大肠癌组织中Skp2与p27kip1表达呈负相关(r=-0.430,P<0.01)。结论大肠癌中Skp2蛋白的过度表达与p27kip1蛋白降解有关,提示Skp2蛋白可能在大肠癌的发生发展中起重要作用。     

Immunohistochemical analysis of TrkA neurotrophin receptor expression in human non-neuronal carcinomas

The Trk family of tyrosine protein kinase receptors plays a significant role in the development and maintenance of neural tissues. It has been recently shown that Trk receptors are also expressed by a wide range of normal non-neuronal tissues in humans in a cell type-specific manner. In the present study, the expression patterns of TrkA in 337 non-neuronal invasive carcinomas of 15 different human tissues were investigated immunohistochemically. Overall, 133 (39%), 101 (30%) and 103 (31%) tumors exhibited strong, moderate and no TrkA Immunoreactivity, respectively. Esophageal and thyroid carcinomas expressed high levels of TrkA, whereas the levels in gastric and colon cancers were low. TrkA expression was detected not only in carcinomas originating from TrkA-positive normal counterpart tissues, Including the esophagus, breast, lung and uterus, but also in those from TrkA-negative tissues/cells of the thyroid, liver and ovary. Immunostaining for nerve growth factor-β, the specific ligand for TrkA, in esophageal and breast carcinomas demonstrated its immunoreactivity in stromal fibroblasts and some TrkA-expressing tumor cells. These results suggest that paracrine/autocrine regulation via stromal/tumoral NGF-tumoral TrkA interaction may be involved In the growth of certain non-neuronal carcinomas.     

白血病细胞p16基因突变分析

目的探讨ｐ１６基因突变在白血病发生中的作用及基因突变的机制。方法利用点突变检测仪、水平和垂直板电泳对ｐ１６基因的外显子１、外显子２的ＰＣＲ扩增产物作缺失和点突变分析。结果在白血病３５例临床标本中有２２例发生缺失突变，６例发生点突变，突变率约８０％。在２２例缺失突变病例中，有１０例为不完全缺失突变即有低于外显子５０９ｂｐ的扩增产物。结论ｐ１６基因含有“ＧＣ”ＤＮＡ重复顺序，易发生ＤＮＡ重组及易位和重排。在白血病发生中起重要作用