Stathmin基因表达与肿瘤的研究进展

Stathmin在多种恶性肿瘤细胞中都有高水平表达，Stathmin蛋白的主要作用是通过促进微管的解聚或阻止微管的聚合从而影响有丝分裂纺锤体的形成，通过抑制其表达可以干扰恶性肿瘤细胞的有丝分裂，影响肿瘤细胞的增殖与凋亡。同时，抑制stathmin表达能够协同增效某些化疗药物的抗癌疗效。Stathmin基因正成为肿瘤基因治疗的一个新靶点。     

Stathmin蛋白的研究进展

Stathmin蛋白由于其特有的微管解聚活性,在细胞的增殖和分化及肿瘤发生中有十分重要的作用.抑制Stathmin蛋白的表达已经成为肿瘤基因治疗的新的靶点,并且已经证实Stathmin蛋白能够影响某些作用于微管的化疗药物的疗效,对于指导临床用药有一定的意义.Stathmin蛋白还能够促进神经系统的发育,因此受到更多的关注.     

抗人微管不稳定蛋白单克隆抗体的制备及鉴定

目的制备人微管不稳定蛋白（Stathmin）的单克隆抗体,检测Stathmin蛋白在真核细胞中的表达,为探讨Stathmin 生物学功能奠定基础。方法利用重组Stathmin蛋白为免疫原,免疫BALB/C小鼠,取免疫小鼠的脾细胞和同系小鼠的骨髓瘤细胞Sp2/0进行常规融合,通过间接ELISA的筛选和有限稀释克隆化,获得稳定分泌抗Stathmin单克隆抗体的杂交瘤细胞株,通过ELISA,Western blot和免疫组织化学实验等方法分别对其效价和特异性进行鉴定。结果成功地建立了2株稳定分泌抗Stathmin的单克隆抗体杂交瘤细胞株,分别命名为F001和F002。两株单克隆抗体的免疫球蛋白亚类均为IgG1。两株单克隆抗体通过免疫组织化学实验和Western blot实验都能特异性地结合真核细胞内源性的Stathmin蛋白。结论成功建立了两株效价高、特异性好的抗Stathmin蛋白的单克隆抗体,为进一步研究Stathmin的生物学功能及其与肿瘤的关系创造了条件。     

Stathmin在上皮性卵巢癌组织中的表达及意义

背景与目的:Stathmin是一种重要的微管调节蛋白,参与多种肿瘤的发生、发展.本研究旨在验证并探讨Stathmin在上皮性卵巢癌中的表达及意义.方法:利用免疫组化和逆转录聚合酶链反应(RT-PCR)方法检测22例卵巢正常上皮组织,16例良性肿瘤和50例上皮性卵巢癌组织中Stathmin的表达.结果:上皮性卵巢癌中Stathmin mRNA的表达明显高于卵巢良性肿瘤(P<0.05)和正常卵巢上皮(PO.05).进一步利用x2检验统计分析得出,Stathmin蛋白与卵巢癌的临床分期、病理分级、淋巴转移相关(P<0.05),而与年龄、病理类型及腹水量无相关性(P>0.05).结论:Stathmin在上皮性卵巢癌中高表达,可能与卵巢癌的发生、发展相关.     

非小细胞肺癌β-tubulin-Ⅲ和Stathmin基因表达与化疗敏感性的研究

目的:探讨Ⅱ期非小细胞肺癌(NSCLC)组织中Ⅲ型β微管蛋白(β-tubulin-Ⅲ)和Stathmin基因mRNA表达与诺维本+顺铂(NP)方案化疗敏感性的关系,为个体化治疗方案的制定提供依据.方法:单基因定量法检测73例已手术Ⅱ期NSCLC患者癌组织中β-tubulin-Ⅲ和Stathmin的mRNA表达.所有患者行NP方案化疗,并随访患者的无瘤生存时间(DFS)和总生存时间(OS).统计学分析β-tubulin-Ⅲ和Stathmin的mRNA表达水平与患者DFS和OS的关系.结果:73例患者的OS与β-tubulin-Ⅲ的mRNA表达水平有关,P=0.003.患者的DFS与β-tubulin-Ⅲ的表达及淋巴结转移情况有关,P＜0.01.绘制生存曲线表明,癌组织中β-tubulin-Ⅲ低表达患者的OS和DFS均高于β-tubulin-Ⅲ高表达者,P＜0.05;Stathmin低表达患者的OS和DFS与高表达患者相比,差异均无统计学意义,P＞0.05.结论:Ⅱ期NSCLC患者癌组织中,β-tubulin-Ⅲ表达水平可以作为预测患者预后及化疗敏感性的指标,可作为选择合适化疗药物的参考.     

Stathmin 基因在非小细胞肺癌中的表达

目的：检测 Stathmin 基因在非小细胞肺癌组织中的表达,探讨其与临床病理因素的关系。方法：用免疫组织化学 SP 法检测135例非小细胞肺癌及20例癌旁肺组织中 Stathmin 蛋白的表达,分析其与患者年龄、性别、肿瘤分化程度等临床病理特征之间的相关性。结果：小细胞肺癌组织中 Stathmin 基因蛋白的表达水平明显高于癌旁肺组织（P ＜0．05）,Stathmin 的表达与淋巴结转移有关（P ＜0．05）。结论：Stathmin 基因在非小细胞肺癌组织中的表达显著上调;Stathmin 基因的表达与淋巴结转移有关,而与年龄、性别、肿瘤大小、TNM分期、组织类型无关。     

微管不稳定蛋白Stathmin在食管鳞癌组织中的表达及意义

目的 探讨Stathmin蛋白在食管鳞癌组织中的表达及意义.方法 收集新乡医学院2006年4月至9月期间食管鳞癌切除标本及其相应癌旁组织31例,以免疫组织化学法检测Stathmin蛋白在所取标本中的表达情况.结果 在31例食管鳞癌标本中,Stathmin蛋白表达阳性的标本为23例,阳性率为74.2%,阳性颗粒可见于细胞质;分别按年龄、性别、组织类型、分化程度、淋巴结转移、肿瘤浸润深度及TNM分期将31例食管癌标本分组,Stathmin蛋白表达差异有统计学意义的因素有分化程度、淋巴结转移、肿瘤浸润深度及TNM分期(P＜0.05).结论 食管鳞癌组织中Stathmin蛋白高表达,其表达与食管鳞癌的分化程度、淋巴结转移、肿瘤浸润深度及TNM分期显著相关,Stathmin蛋白可能为人食管癌的生物治疗提供一个新靶点.     

Stathmin蛋白:一个潜在的肿瘤标志物

人stathmin也称原癌基因蛋白18(oncogene protein 18, Op18), 是一种广泛存在于细胞质的蛋白质,其相对分子质量约为19×10~3,可与微管蛋白结合,参与微管和纺锤体的组装;与细胞的增殖、分化、再生和运动均有关,并具有信号活性调节的功能.近年来有研究发现,stathmin在多种肿瘤中高表达,并可通过调节微管的解聚,促进肿瘤细胞的运动及侵袭.Stathmin翻译后修饰状态的改变,可影响与p53蛋白的相互作用,参与肿瘤的发生发展.目前单独或者联合化疗药物使用的抗stathmin效应剂已用于某些肿瘤的治疗.虽然stathmin与肿瘤病因学的内在联系尚不十分清楚,但其作为一个潜在的肿瘤标志物或药物靶点值得进一步研究探讨.     

Stathmin在胃癌中的表达及其与多西他赛疗效的关系

摘 要：［目的］ 探讨Stathmin在胃癌中的表达及与多西他赛化疗疗效关系。［方法］ 应用免疫组化法分析Stathmin在胃癌及癌旁组织中的表达情况,研究其与胃癌临床病理特征的关系;研究Stathmin表达与术后辅助DC（多西他赛+顺铂）方案化疗有效率、临床获益率、1年生存率、3年生存率之间的关系。［结果］ 胃癌组织Stathmin表达较癌旁组织增高(P＜0.05),与胃癌组织分化程度、浸润深度、有无淋巴结转移及临床分期均相关(P＜0.05),而与患者的年龄、性别无关;Stathmin过表达较低表达患者化疗敏感性下降(P＜0.05),1年生存率、3年生存率尚无统计学意义(P＞0.05)。［结论］ Stathmin表达与胃癌组织的生物学行为相关;胃癌组织中Stathmin蛋白的表达与患者对含多西他赛化疗方案的敏感性有关,但对生存时间的影响证据尚不足。     

Stathmin蛋白在大肠癌中的表达及临床意义

目的:探讨Stathmin蛋白在大肠癌中的表达及其与肿瘤病理特征的关系.方法:采用免疫组织化学SP法检测56例大肠癌组织Stathmin蛋白的表达,同时选取16例大肠腺瘤组织作对照.结果:Stathmin蛋白在大肠癌中的阳性表达率为60.7%(34/56),明显高于大肠腺瘤组织的31.3%(5/16),P <0.05.Stathmin蛋白表达与肿瘤部位、大小、分化程度、Duke's临床分期及淋巴结转移均无明显相关,P >0.05.结论:Stathmin蛋白过表达与大肠癌的发生密切相关,但它不能作为判断大肠癌恶性程度的指标.     

Silencing stathmin gene expression by survivin promoter-driven siRNA vector to reverse malignant phenotype of tumor cells

Stathmin gene overexpression has been shown to play an important role in maintenance of malignant phenotype in tumor cells, and the blocking efficacy and tumor specificity of this target has been concerned in clinical trails. In this report, we designed survivin promoter-driven siRNA eukaryotic expression vector that expressed the small interfering RNA targeting stathmin gene to selectively knock down the stathmin gene expression in two different kinds of tumor cell lines while sparing normal cell lines. The therapeutic potential of this recombinant vector was tested in human cervical cancer Hela cells and osteosarcoma SSOP-9607 cells, and in human umbilical vein endothelial cell line ECV304 cells as control. The siRNA vector- transfected Hela cells and SSOP-9607 cells revealed marked inhibition of stathmin expression and a dramatic growth inhibition comparing with ECV304 cells, parental-vector transfected cells and untransfected cells. Cell cycle analysis of siRNA vector transfected tumor cells by Flow Cytometry showed G(2)/M phase block, while morphologic analysis by TURNEL staining method showed marked increase of apoptosis. Our study indicates that survivin gene promoter-driven stathmin siRNA expression vector may have potential use in tumor gene therapy with targeted tumor gene silencing effect.     

stathmin基因SiRNA对人宫颈癌细胞的生长抑制作用

目的探讨S iRNA封闭stathm in基因表达对人宫颈癌细胞生长抑制作用。方法用Amb ion公司pS ilencer4.1CMV构建针对stathm in基因的S iRNA真核表达载体,以高表达stathm in基因的人宫颈癌Hale细胞系为靶细胞,脂质体法转染S iRNA表达载体,RT-PCR分析转染细胞stathm in基因表达;MTT试验观察stathm in S iRNA对Hale的生长作用影响;流式细胞仪分析细胞增殖周期的改变。结果stathm in S iRNA可有效封闭stathm in基因表达且明显抑制了宫颈癌Hale细胞的生长;Hale细胞在stathm in S iRNA作用下,细胞分裂阻滞在分裂期的中期,并诱导细胞发生凋亡。结论stathm in基因对宫颈癌细胞的生长可能起十分重要的作用,它有望成为宫颈癌治疗的新靶点。     

Effect of stathmin on the sensitivity to antimicrotubule drugs in human breast cancer

Stathmin is a p53-regulated protein known to influence microtubule dynamics. Because several chemotherapeutic agents used to treat breast cancer alter the dynamic equilibrium of tubulin polymerization, stathmin may play an important role in determining the sensitivity to these drugs. Therefore, we evaluated the effect of stathmin expression on the action of taxanes and Vinca alkaloids using a panel of human breast cancer cell lines. Cell lines harboring mutant p53 expressed high levels of stathmin. Two cell lines with different levels of endogenous stathmin expression and isogenic-paired cell lines transfected to overexpress stathmin were used to determine whether or not stathmin modulated the sensitivity to drugs. Overexpression of stathmin decreased polymerization of microtubules, markedly decreased binding of paclitaxel, and increased binding of vinblastine. Stathmin overexpression decreased sensitivity to paclitaxel and, to a lesser extent, to vinblastine. In contrast, stathmin content had no significant effect on the sensitivity to chemotherapeutic drugs that do not target microtubules. Cell lines overexpressing stathmin were more likely to enter G(2) but less likely to enter mitosis as determined by fluorescence-activated cell sorting and mitotic index. This effect was magnified when stathmin-overexpressing cells were treated with vinblastine as measured by the detection of proteins phosphorylated in early mitosis. These data suggest that the action of antimicrotubule drugs can be affected by stathmin in at least two ways: (a) altered drug binding; and (b) growth arrest at the G(2) to M boundary. Mutant p53 breast cancers exhibiting high levels of stathmin may be resistant to antimicrotubule agents.     

The phosphoprotein Op18/stathmin is differentially expressed in ovarian cancer

To identify potential prognostic indicators of ovarian cancer and identify targets for therapeutic strategies, mRNA differential display was used to analyze gene expression differences in normal, benign, and cancerous ovarian tissue. One cDNA isolated by this technique, Op18/stathmin, is a highly conserved gene that is reported to have many different functions within a cell, including signal transduction, control of the cell cycle, and the regulation of microtubules. Quantitative Northern blot analysis of 12 malignant ovarian samples, 8 benign ovarian tumors, and 10 normal ovarian tissue samples demonstrated overexpresaion of Op18/stathmin mRNA in the malignant cancers. Immunohistochemistry showed an apparent overex-pression of Op18/stathmin protein level and an association with proliferating cells     

Stathmin基因在人成骨肉瘤细胞中的表达及意义

stathmin作为细胞信号转导分子在细胞分化及恶性肿瘤发生、发展上发挥重要作用。本研究旨在探讨Stathmin基因在人成骨肉瘤细胞中的表达,并探讨阻断其表达对该肿瘤细胞的生物学行为的影响。方法：RT-PCR及原位杂交法检测2个人成骨肉瘤细胞系及45例人成骨肉瘤组织中Stathmin基因表达情况;以高表达Stathmin基因的人成骨肉瘤细胞系SOSP-9607为靶细胞、反义Stathmin(ASODN)为阻断剂,通过MTT实验观察ASODN对该细胞系的生长抑制作用,并用流式细胞仪分析其对细胞增殖周期的影响。     

Gene expression profiling in the mammary gland of rats treated with 7,12-dimethylbenz[a]anthracene

Identification of molecular markers of early-stage breast cancer development is important for the diagnosis and prevention of the disease. In the present study, we used microarray analysis to examine the differential expression of genes in the rat mammary gland soon after treatment with a known chemical carcinogen, 7,12-dimethylbenz[a]anthracene (DMBA), and prior to tumor development. Six weeks after DMBA, differential expression of multiple genes involved in cell growth, differentiation and microtubule dynamics were observed. Gene expression changes were further validated by a combination of techniques, including real-time PCR, RT-PCR, Western blotting and immunohistochemistry. An inhibition of differentiation in this early stage was suggested by the lower expression of beta-casein and transferrin and higher expression of hsp27 in glands from DMBA-treated rats. Possible cell cycle deregulation was indicated by an increased expression of cyclin D1 and hsp86, a heat shock protein associated with cyclin D1. Prior to tumor development, DMBA increased cellular proliferation as detected by Ki-67 and stathmin immunostaining in histologically normal mammary gland. Genes regulating microtubule function, including stathmin, Ran, alpha-tubulin and hsp27, were all overexpressed in the mammary gland of DMBA-treated rats, raising the possibility that disruption of microtubule dynamics and abnormal mitosis may be critical events preceding breast cancer development. Several of the altered proteins, including hsp27, hsp86 and stathmin, may ultimately serve as markers of early breast cancer development.     

靶向stathmin和mdr1基因逆转卵巢癌细胞 紫杉醇耐药的研究

 目的探讨以RNA干扰（RNA interference,RNAi）技术靶向stathmin和mdr1基因逆转卵巢癌细胞紫 杉醇耐药的可行性。方法分别构建靶向stathmin和mdr1基因的质粒：pGU6-GFP-neo-STMN1和pGU6-GFP- neo-MDR1;将质粒转染到卵巢癌紫杉醇耐药细胞株SKOV3/TAX。 Real-time RT-PCR检测stathminm和mdr1 的mRNA变化;Western blot检测其蛋白表达变化;荧光显微镜检测细胞凋亡情况;CCK-8法分析细胞对紫 杉醇的敏感度。结果Real-time RT-PCR及Western blot显示pGU6-GFP-neo-MDR1质粒对mdr1基因, pGU6- GFP-STMN1-294shRNA质粒对stathmin基因在mRNA水平和蛋白水平均有明显抑制（P<0.05）, 荧光显微镜 下共转染组细胞凋亡增多,CCK-8示共转染组逆转紫杉醇耐药效果最明显。结论体外RNAi可有效沉默卵巢 癌紫杉醇耐药株SKOV3/TAX细胞内stathmin基因和mdr1基因,逆转其紫杉醇耐药。