The Research and Development of Gene Therapy for Breast Cancer by Src Signal Tiansduction Pathways

背景：SRC Homology 2结构域蛋白酪氨酸磷酸酶底物-1(SHPS-1 TS，又称P84、BIT、SIRP)是一种参与受体酪氨酸激酶信号转导的异嗜性粘附膜蛋白，广泛表达于哺乳动物的中枢神经系统、免疫系统等组织中。SHPS-1的功能是控制细胞的迁移和扩散。我们研究了SHPS-1的细胞定位及其与包括预后在内的临床病理特征的关系。设计：采用免疫组织化学和Western blotting方法检测5个细胞系和63例乳腺癌标本中SHPS-1的表达和细胞定位。所用的单抗为SHPS-1(兔多克隆抗体，Neomarkers)。研究的临床病理因素包括年龄、分期、肿瘤类型、组织学分级和核分裂指数(MI)。用Kaplan-Meier方法评估总生存率和无病生存率。结果：通过Western blotting分析，在组织样本和细胞系中检测到100 kDa的蛋白。在正常粘膜上皮细胞中有膜性表达。乳腺癌中胞浆阳性(c+)22例(35%)，阴性4例(65%)，c(+)与c(-)在年龄、分期、类型等方面无明显差异。SHPS-1的表达与组织学分级有显著相关性：c+21例(95%)为G2-G3级(P&lt；0.05)。C(+)组MI(1.86±1.6)显著高于C(-)组(0.71±0.56)。C(+)患者的总体生存率和无病生存率均低于c(-)患者(P=0.044和P=0.015)。结论：SHPS-1胞浆表达可能与基因突变有关。SHPS-1可能是基因治疗的合理候选者。

Background : Src homology 2 domain-containing protein tyrosine phosphatase(SHP) substrate-1(SHPS-1 TS also known as p84,BIT, and SIRP) is a heterophilic adhesive membrane protein involved in receptor tyrosine kinase signaling that is expressed widely in the mammalian central nervous system, immune system and other tissues. The function of SHPS-1 is to control cell migration and spreading.We studied the SHPS-1 cellular localization and its significance in relation to clinicopathological features including prognosis.Design : Five cell lines and 63 specimens of breast carcinomas were examined to evaluate SHPS-1 expression and cellular localization by immunohistochemistry and western blotting. The monoclonal antibodies used was SHPS-1 (rabbit polyclonal antibody, Neomarkers). The clinicopathological factors studied were age, staging, tumor type, histological grade and mitotic index(MI). Overall survival and diseases-free survival were evaluated using the Kaplan-Meier method.Result : We detected 100-kDa proteins in tissue samples and cell lines by Western blotting analysis. In normal epithelium, there was a membranous expression. In breast carcinomas, 22 cases (35%) showed cytoplasmic positivity (c+), whereas 4 leases (65%) were negative (c-), There was no difference in age, staging, type between c(+) and c(-). There was a statistically significant correlation between SHPS-1 expression and histological grade : 21cases (95%) of c(+) were G2-G3 tumors (P<0.05). The MI for c(+) cases(1.86±1.6) was significantly higher than that for C(-)(0.71±0.56). The cases with c(+) had poor overall and diseases-free survival rates than c(-)(P=0.044 and P=0.015,respectively).Conclusion : It was suggested that cytoplasmic expression of SHPS-1 could be associated with gene mutation. SHPS-1 could be the logical candidate for gene therapy.