野生型p53基因诱导凋亡相关基因ANNEXIN A2的表达研究

目的探讨胡基因过表达介导不同转移潜能肿瘤细胞的凋亡分子机理，寻找与细胞凋亡和肿瘤转移的相关基因。方法应用mRNA差异显示方法分析腺病毒介导的野生型,053基因感染不同转移潜能的肺癌细胞系前后存在的表达差异基因，并用逆转录一聚合酶链反应、Northern印迹和Western印迹方法加以验证。结果分析发现，在柳基因诱导后ANNEXIN A2基因的表达显著差异有统计学意义，经加基因诱导后ANNEXIN A2基因的表达有明显下调。并且以Arfip 973细胞的表达缺失最为显著。经逆转录一聚合酶链反应、Northern印迹和Western印迹方法也验证了这种差异的存在。结论表明ANNEXIN A2基因的表达与p53基因诱导的肿瘤细胞凋亡存在密切的相关性。     

Inflammatory Neutrophils Secrete Annexin 1 During Experimentally Induced Colitis in Rats

Immunoblotting and immunohistochemical analysiswere performed to identify the cells expressing andsecreting annexin 1 during experimental rat colitisinduced by trinitrobenzene sulfonic acid. Annexin 1 expression was increased during theinflammation. Likewise, annexin 1 secretion was inducedin inflamed colons at one, three, six, and nine daysafter trinitrobenzene sulfonic acid treatment but wasnot detected in colons from controls and rats at 12days. Immunohistochemistry showed that the rise inannexin 1 expression resulted from the infiltration oftwo types of leukocytes highly positive for annexin 1: neutrophils (the most abundant) andmacrophages. At day 1 after treatment, neutrophils ofthe inflammatory site, in mucosa and submucosa, are theonly cells expressing annexin 1. Immunoblotting showed that they secreted annexin 1 whereasneutrophils from blood or tunica muscularis did not.This indicates that, during this colitis, annexin 1 canbe secreted by neutrophils located in the inflammatorysite.     

Depletion of annexin A5, annexin A6, and collagen X causes no gross changes in matrix vesicle–mediated mineralization,but lack of collagen X affects hematopoiesis and the Th1/Th2 response

Numerous biochemical studies have pointed to an essential role of annexin A5 (AnxA5), annexin A6 (AnxA6), and collagen X in matrix vesicle–mediated biomineralization during endochondral ossification and in osteoarthritis. By binding to the extracellular matrix protein collagen X and matrix vesicles, annexins were proposed to anchor matrix vesicles in the extracellular space of hypertrophic chondrocytes to initiate the calcification of cartilage. However, mineralization appears to be normal in mice lacking AnxA5 and AnxA6, whereas collagen X–deficient mice show only subtle alterations in the growth plate organization. We hypothesized that the simultaneous lack of AnxA5, AnxA6, and collagen X in vivo induces more pronounced changes in the growth plate development and the initiation of mineralization. In this study, we generated and analyzed mice deficient for AnxA5, AnxA6, and collagen X. Surprisingly, mice were viable, fertile, and showed no obvious abnormalities. Assessment of growth plate development indicated that the hypertrophic zone was expanded in Col10a1^?/? and AnxA5^?/?AnxA6^?/?Col10a1^?/? newborns, whereas endochondral ossification and mineralization were not affected in 13‐day‐ and 1‐month‐old mutants. In peripheral quantitative computed tomography, no changes in the degree of biomineralization were found in femora of 1‐month‐ and 1‐year‐old mutants even though the diaphyseal circumference was reduced in Col10a1^?/? and AnxA5^?/?AnxA6^?/?Col10a1^?/? mice. The percentage of naive immature IgM⁺/IgM⁺ B cells and peripheral T‐helper cells were increased in Col10a1^?/? and AnxA5^?/?AnxA6^?/?Col10a1^?/? mutants, and activated splenic T cells isolated from Col10a1^?/? mice secreted elevated levels of IL‐4 and GM‐CSF. Hence, collagen X is needed for hematopoiesis during endochondral ossification and for the immune response, but the interaction of annexin A5, annexin A6, and collagen X is not essential for physiological calcification of growth plate cartilage. Therefore, annexins and collagen X may rather fulfill functions in growth plate cartilage not directly linked to the mineralization process. © 2012 American Society for Bone and Mineral Research.     

食管鳞状上皮癌中表达失调蛋白的研究

背景与目的：食管鳞状上皮癌（esophageal squamous cell carcinoma,ESCC)是一种具有高发性和高死亡率疾病,本研究应用蛋白质组学方法分析ESCC中异常表达的蛋白。方法：对ESCC／癌旁组织标本的冰冻切片进行显微切割,分别分离肿瘤细胞和癌旁上皮细胞,提取细胞总蛋白,通过双相电泳得到ESCC及癌旁上皮的蛋白表达谱,采用质谱技术鉴定肿瘤表达异常的蛋白。结果：比较7对食管癌／癌旁组织和蛋白表达谱,并对差异的蛋白进行质谱分析发现：11个蛋白在食管癌中表达明显异常,其中热休克蛋白27（HSP27）在3例ESCC中表达上调,钙依赖磷脂结合蛋白ANNEXINI尖正常食管粘膜存在分子量基本相同、等电点彼此相差约为1的3种变体（isoforms),3种ANNEXINI变体均在5例食管癌中表达下调。结论：双相电泳连接质谱的蛋白质组学方法是快速筛选肿瘤异常表达蛋白的有效手段,HSP27和ANNEXINI的表达失调是食管癌发生过程中的频发事件。