siRNA沉默整合素beta1 基因对子宫内膜癌细胞侵袭、转移的影响

目的：探讨siRNA 沉默整合素茁1 基因对子宫内膜癌细胞侵袭、转移的影响。方法：选取子宫内膜癌ECC-1细胞系(ER阳性) 和KLE细胞系(ER阴性)，分别转染Integrin beta1 siRNA 质粒(Integrin beta1 siRNA 组)、无义序列siRNA质粒(无义序列对照组)和空载 质粒(空载对照组)，利用实时荧光定量PCR 检测各组细胞中Integrin 茁1 mRNA的表达，Western blot 检测各组细胞中Integrin beta1、 beta-catenin 和C-Myc 蛋白的表达，Transwell 小室检测各组细胞迁移和侵袭能力，MTT 法检测各组细胞的增殖情况。结果：Integrin beta1 siRNA 组ECC-1 细胞和KLE 细胞中Integrin beta1 mRNA 和蛋白相对表达量均低于无义序列对照组和空载对照组(P<0.05)； Integrin beta 1 siRNA组ECC-1 细胞和KLE细胞中beta-catenin 蛋白和C-Myc 蛋白相对表达量均低于无义序列对照组和空载对照组， 差异均有统计学意义(P<0.05)；Integrin beta1 siRNA组ECC-1 细胞和KLE 细胞中迁移细胞数和侵袭细胞数均低于无义序列对照组 和空载对照组(P<0.05)；Integrin beta1 siRNA 组ECC-1细胞和KLE 细胞的A 值均低于无义序列对照组和空载对照组(P<0.05)。结 论：特异性抑制Integrin beta1 基因可抑制子宫内膜癌细胞迁移、侵袭和增殖，可能与抑制Wnt信号传导有关。     

癌基因Src在骨肉瘤细胞侵袭伪足形成中的作用

目的:探讨癌基因Src在体外培养骨肉瘤细胞侵袭伪足形成中的作用。方法:构建Src sh RNA慢病毒表达载体,在HEK293T细胞中包装慢病毒,感染HT-1080骨肉瘤细胞,经嘌呤霉素加压筛选,获得稳定沉默Src基因的骨肉瘤细胞系HT-1080-sh Src;实时定量PCR和Western Blot法检测基因沉默效率;采用原位明胶酶谱法检测侵袭伪足形成;采用侵袭小室实验检测下调Src基因表达对HT-1080细胞侵袭力的影响。结果:成功构建稳定沉默Src基因的骨肉瘤细胞系HT-1080-sh Src及对照细胞系HT-1080-shluc,经实时定量PCR和Western Blot检测,与对照细胞系相比,HT-1080-sh Src细胞中Src基因表达下调3倍以上;下调HT-1080细胞中Src基因表达能显著抑制HT-1080细胞侵袭伪足形成及其对细胞外基质的降解能力;下调Src基因表达能显著抑制骨肉瘤细胞侵袭力。结论:癌基因Src参与调节骨肉瘤细胞HT-1080侵袭伪足形成,促进肿瘤侵袭、转移。     

植物基因转移方法

基因工程的诞生和发展是与基因转移方法的出现和发展分不开的。为了实现不同的目标，就需要各种各样的基因转移方法。一般来说，向植物中转移基因，存在的困难要比微生物和动物多些；但由于植物基因工程对作物改良的重要性，近年来获得了迅速发展，各种基因转移方法层出不穷。本文系统地介绍了现有的植物基因转移方法。     

蛋白质截短检测在基因突变在检测中的应用

蛋白质截短检测（ｐｒｏｔｅｉｎ ｔｒｕｎｃａｔｉｏｎ ｔｅｓｔ，ＰＴＴ）是从蛋白质水平对基因突变进行检测的方法。由于ＰＴＴ具有与以往广泛使用的方法不同的独特优点，自１９９３年发明以来，先后在ＡＰＣ、ＤＭＤ、ＢＲＣＡ１、错配修复基因（ｍｉｓｍａｔｃｈ ｒｅｐａｉｒ ｇｅｎｅ）突变检测中得以应用。     

建立以TK基因为标记将外源基因导入臭曲霉（Asper—gillus...

A new system for selection of transformed Aspergillus foetidus was reported. In this system, TK- A. foetidus which were constructed by homologous recombination of mutated TK gene of vaccinia virus with TK gene of A. foetidus were screened by adding BUdR in agar plates. Conditions for screen of TK+ A. foetidus strain, transformation of A. foetidus and selection of transformed TK- A. foetidus have been studied. By using this system, several transformed A. foetidus which contained HBsAg gene derived bf a promoter H8 cloned from genomic DNA of A. foetidus were isolated. It was demonstrated that HBsAg gene was integrated into the chromosome DNA of A. foetidus by Southern blot after many passages of spores. ELISA showed that HBsAg was positive in the growth medium (p/n = 20). The 22 nm particles which were very similar to the HBsAg particles in human serum were found in the growth medium by immunoelectromicroscope. Western blot also gave the specific bands. All these data showed that HBsAg gene was expressed in A. foetidus and the products were secreted into the growth medium. The selection system using TK gene as marker could generally be used to study the expression of foreign gene in A. foetidus.