突触核蛋白-γ对乳腺癌雌激素受体表达的影响

目的探讨突触核蛋白-7（synuclein-7,SNCG）表达对乳腺癌雌激素受体（estrogen receptor,ER）表达的影响。方法构建慢病毒SNCG过表达及干扰栽体,分别转染人乳腺癌T47D（SNCG＋／ER＋）细胞株。在SNCG过表达和SNCG干扰细胞组中,RT-PCR方法栓泖lSNCG和ER的rflRNA水平表达,Western blot法检测SNCG和ER的蛋白水平表达,并做Pearson法相关统计分析。结果在mRNA水平,SNCG过表达组中ER基因的mRNA表达量显著增高,两者呈正相关（P=0．03,R=0．81）;SNCG干扰组中ER基因的mRNA表达量显著降低,两者呈正相关（P=0．01,R=0．82）。在蛋白表达水平,SNCG过表达组ER表达显著增强,两者呈正相关（P=0．03,R-0．85）;SNCG干扰组ER表达显著降低,两者呈正相关（P-0．04,R=0．83）。结论SNCG对ER表达有正向调节作用,为进一步研究SNCG与ER的相互作用机制及SNCG与乳腺癌内分泌治疗之间的关系奠定了实验基础。     

ADM与5Fu可抑制MDA-MB231乳癌细胞的SNCG表达

目的 研究乳癌临床常用化疗药物顺铂(ciaplalin or DDP),阿霉素(adriamycin,ADM),氟尿嘧啶(fluorouradl,5Fu)对MDA-MB231乳癌细胞SNCG表达的干扰效应.方法 通过RT-PCR及免疫组织化学法检测上述药物处理组和阴性对照组MDA-MB231细胞的SNCG表达状况,用Quantity One软件及北航真彩色医学图像处理系统(CM-20008)分别对各组SNCG mR-NA和蛋白相对表达水平进行分析.结果 ADM和5Fu处理组与阴性对照组比较,MDA-MB231细胞的SNCG mRNA及蛋白表达水平差异均有统计学意义(P值均小于0.05),而DDP处理组表达水平与对照组无差别.结论 ADM和5Fu可抑制MDA-MB231细胞的SNCG表达.     

Expression of SNCG,MAP2, SDF-1 and CXCR4 in gastric adenocarcinoma and their clinical significance

Objectives: The purpose of the study was to detect the expression of SNCG, MAP2, SDF-1 and CXCR4 in gastric adenocarcinoma, and to evaluate their roles in the carcinogenesis of gastric adenocarcinoma, development, invasion and metastasis as well as their clinical significance. Methods: The expression of SNCG, MAP2, SDF-1 and CXCR4 was detected by SP immunohistochemical method in 225 cases of gastric adenocarcinoma and 105 cases of nonneoplastic adjacent gastric tissue. The expression of SNCG, MAP2, SDF-1 and CXCR4 mRNA was also detected by RT-PCR method in 50 cases of gastric adenocarcinoma and 30 cases of nonneoplastic adjacent gastric tissue. Results: The expression of SNCG, MAP2, SDF-1 and CXCR4 in the gastric adenocarcinoma was remarkably higher than those in the nonneoplastic adjacent gastric tissue (P < 0.01); The positive expression of SNCG and MAP2 was correlated with the depth of tumor invasion and the metastasis of lymph nodes (P < 0.05), and that of SDF-1 and CXCR4 was correlated with the metastasis of lymph nodes (P < 0.05). Conclusions: SNCG, MAP2, SDF-1 and CXCR4 may play an important role in the carcinogenesis, progression, invasion and metastasis of gastric adenocarcinoma. However, it still needs more exploration whether they can serve as promising therapeutic targets of gastric adenocarcinoma.     

Alternative splicing of synuclein gamma in endometrial cancer: identification of a novel isoform

Synuclein gamma (SNCG) is under consideration as a potential biomarker in cancer biology. Up to date four different SNCG variants are described. Due to growing evidence suggesting correlations between aberrant alternative splicing processes and cancer progression, we investigated the effects of peritumoural conditions on expression pattern of SNCG in endometrial cancer (EC) in vitro. Compared to breast cancer cell lines, mRNA expression levels of all known SNCG isoforms 1–4 are significantly reduced in EC cell lines. We identified a novel alternatively spliced variant of isoform 2 (isoform 2 short) which is found highly expressed in EC cell lines. Hypoxia and acidosis trigger an up-regulation of isoform 2 short. EC cell lines are characterized by low SNCG protein levels under control conditions, but exhibit a significant increase triggered by hypoxia and acidosis. In addition we analysed the potential association between SNCG protein expression and clinico-pathological parameters in human EC samples. Our findings indicate a grade-dependent induction of SNCG protein expression in endometrial cancer.We identified for the first time a novel isoform of SNCG that is found specifically expressed in EC. Our results also strongly indicate the existence of a corresponding protein of isoform 2 short that potentially plays a critical role in EC cancer progression.     

SNCG在肝癌高/低淋巴道转移细胞株中的差异性表达

目的:研究SNCG蛋白在小鼠肝癌高/低淋巴道转移潜能细胞株中的定位和表达差异。方法:应用免疫细胞化学、Western blot方法检测SNCG蛋白在小鼠肝癌高/低淋巴道转移潜能细胞株Hca-F/Hca-P的定位情况和表达差异。结果:SNCG蛋白在Hca-F和Hca-P细胞株中均主要定位于细胞浆,细胞核区域也可见少量蛋白表达。SNCG在Hca-F中的表达［染色强度为(+++)］强于在Hca-P中的表达［染色强度为(++)］。检测到一条SNCG特异性蛋白条带,在Hca-F细胞中表达量为Hca-P细胞中的1.9倍。结论:SNCG的高表达可能与高淋巴道转移潜能有关。     

SNCG、 nm23在胃癌中的表达及临床意义

目的:研究胃癌中SNCG,nm23的表达及其临床意义,并进一步探讨它们在胃癌中表达的相关性.方法: 采用免疫组化法观察60例胃癌及20例胃正常黏膜石蜡标本中SNCG、 nm23的表达情况,实验数据用SPSS12.0软件包进行统计.结果: 在胃癌中SNCG蛋白阳性率65%,高于周围正常组织(P<0.05).nm23在胃癌组织中表达阳性率为55%,低于周围正常组织(P<0.01).SNCG、 nm23的表达与肿瘤浸润深度、淋巴结转移及肿瘤临床分期均相关(P<0.05).SNCG与nm23在胃癌中的表达呈负相关(P<0.05)).结论: SNCG、 nm23蛋白的表达与胃癌发生、发展和浸润转移相关.胃癌组织中SNCG的表达与nm23的表达呈负相关,二者的检测有助于胃癌浸润转移的预测,对判断患者预后有重要参考价值.     

siRNA-SNCG增加乳腺癌细胞放射敏感性的实验研究

目的:研究siRNA-SNCG(Synuclein gamma)是否具有增加乳腺癌MDA-MB231细胞株放射敏感性的作用,并探讨可能卷入的分子机制.方法:siRNA特异性敲低MDA-MB231乳腺癌细胞中SNCG表达,使用CCK8细胞毒性实验、7AAD-A/PE-A凋亡实验和Western blot实验检测2 Gy放射作用下对细胞的生物学行为的影响.结果:siRNA特异性敲低SNCG蛋白表达显著增加2 Gy放射对MDA-MB231的细胞毒性和细胞凋亡.在siRNA-SNCG阳性转染组中,Bcl-2蛋白表达受到显著抑制,尤其是在联合放射的情况下,抑制作用更加显著(P<0.001).结论:siRNA-SNCG通过降低Bcl-2表达来增加放射对MDA-MB231的细胞毒性和细胞凋亡,表明抑制SNCG表达具有增加乳腺癌肿瘤细胞放射敏感性的作用.     

siRNA-Mediated Suppression of Synuclein γ Inhibits MDA-MB-231 Cell Migration and Proliferation by Downregulating the Phosphorylation of AKT and ERK

Purpose

Synuclein-γ (SNCG), which was initially identified as breast cancer specific gene 1, is highly expressed in advanced breast cancers, but not in normal or benign breast tissue. This study aimed to evaluate the effects of SNCG siRNA-treatment on breast cancer cells and elucidate the associated mechanisms.

Methods

Vectors containing SNCG and negative control (NC) siRNAs were transfected into MDA-MB-231 cells; mRNA levels were determined by real-time polymerase chain reaction. Cell proliferation was evaluated using the MTT assay, cell migration was assessed by the Transwell assay, apoptosis and cell cycle analyses were conducted with the flow cytometer, and Western blot analysis was performed to determine the relative levels of AKT, ERK, p-AKT, and p-ERK expression.

Results

SNCG mRNA levels were significantly reduced in MDA-MB-231 cells transfected with SNCG siRNA. Our results indicate that in SNCG siRNA-treated cells, cell migration and proliferation decreased significantly, apoptosis was induced, and the cell cycle was arrested. Western blot analysis indicated that the protein levels of p-AKT and p-ERK were much lower in the SNCG siRNA-treated groups, than in the control and NC groups.

Conclusion

SNCG siRNA could decrease the migration and proliferation of breast cancer cells by downregulating the phosphorylation of AKT and ERK.     

γ-神经突触核蛋白在电针改善大脑中动脉闭塞模型大鼠认知功能中的作用

目的:观察电针神庭、百会穴对大脑中动脉闭塞模型(MCAO)大鼠神经功能及海马中γ-神经突触核蛋白(SNCG) mRNA表达的影响,探讨SNCG与记忆认知能力之间的关系。方法:将18只SD大鼠平均随机分为电针组、模型组、假手术组。采用Longa改良线栓法制作MCAO大鼠模型,造模后24h后电针组每日电针神庭、百会穴20min,连续7d,模型组、假手术组予同等条件抓取。通过平衡木实验判断大鼠神经功能缺损状况;实时定量荧光PCR(q-PCR)检测脑组织SNCG mRNA表达。结果:电针组神经功能缺损评分大于假手术组,小于模型组;电针组海马中SNCG mRNA的表达大于假手术组,模型组海马中SNCG mRNA的表达小于假手术组,差异均有统计学意义。结论:电针能改善MCAO大鼠的神经功能状况,SNCG可能与认知功能神经系统之间存在相互作用。