SignalSilence? RCAS1 siRNA I

• 产品编号：CST-6463S      品牌：CST       原厂货号：6463S
• 产品分类：DNA和RNA纯化 > RNA干扰 > RNAi Oligos > siRNA
• 应用分类：

描述：

Cell Signaling Technology (CST)公司的SignalSilence® RCAS1 siRNA I允许研究者通过RNA干扰去特异性抑制RCAS1蛋白表达，这种方法可以通过将双链RNA分子传递到细胞内从而使基因表达有选择的沉默。来自CST的所有的SignalSilence^® siRNA产品都是经过内部严格检测的，并且通过Western blot 分析证明确实能够减少目的蛋白的表达。通过三苯甲基分析每个碱基以监测寡核苷酸的合成，确保合适的配对效率。随后寡核苷酸通过亲和固相萃取法纯化。退火的RNA双链通过质谱分析来证实其精确的组成。每一批产品都通过质谱分析与前面的产品进行比较，来保证不同批次之间的最大一致性。CST公司推荐在细胞裂解前48到72小时转染100 nM RCAS1 siRNA I。转染步骤按照转染试剂说明书提供的步骤进行。遇到任何使用方面的问题，请随时联系CST。SiSo细胞表达的结合受体的癌症抗原(RCAS1)也称雌激素受体结合片段相关基因9(EBAG9)起初被鉴定是一个雌性激素诱导基因(1)，最近研究发现通过负调节细胞毒性T淋巴细胞（CTLs）的杀伤活性，其在适应性免疫应答中扮演一个新的角色(2)。RCAS1基因在系统进化中具有保守性，在许多人源组织和细胞中广泛表达 (3,4)。有证据表明在不同的恶性肿瘤中RCAS1基因的组织表达水平增高，包括胃肠道癌、肝癌、肺癌、乳腺癌、卵巢癌、子宫内膜癌和宫颈癌。此外，RCAS1基因组织表达水平与上述恶性肿瘤患者的预后呈负相关 (4)。同样值得注意RCAS1蛋白升高的水平在癌症患者的血清中已经被检测(4)。初步研究显示RCAS1蛋白是癌细胞分泌的，同时作为T淋巴细胞和B淋巴细胞以及NK细胞上推测的受体的配体，而该配体可诱导这些细胞的凋亡，因此RCAS1可使癌细胞能逃避免疫监视(5,6)。随后研究已经证实RCAS1蛋白作为一个Ⅲ型跨膜高尔基体蛋白，具有调节滤泡的形成、分泌和蛋白质糖基化的功能 (2,7-9)。的确，研究已经证明通过从高尔基体反面膜囊到分泌溶酶体的蛋白运输的负性调节，RCAS1 蛋白的过表达从而负调节CTLs细胞的杀伤功能(2)。此外，RCAS1蛋白过表达可延迟囊泡从内质网到高尔基体的转运，并且使内质网质量控制的组分的和糖基化机器错误定位。因此，RCAS1蛋白可诱导细胞表面上肿瘤相关糖链抗原的沉积，这被认为通过介导黏附、侵袭和转移的而参与肿瘤发生 (8,9)。

1. Watanabe, T. et al. (1998) Mol Cell Biol 18, 442-9.
2. Rüder, C. et al. (2009) J Clin Invest 119, 2184-203.
3. Tsuchiya, F. et al. (2001) Biochem Biophys Res Commun 284, 2-10.
4. Giaginis, C. et al. (2009) Histol Histopathol 24, 761-76.
5. Matsushima, T. et al. (2001) Blood 98, 313-21.
6. Nakashima, M. et al. (1999) Nat Med 5, 938-42.
7. Reimer, T.A. et al. (2005) BMC Cancer 5, 47.
8. Wolf, J. et al. (2010) FASEB J 24, 4000-19.
9. Engelsberg, A. et al. (2003) J Biol Chem 278, 22998-3007.

原厂资料：

Description

SignalSilence® RCAS1 siRNA I from Cell Signaling Technology (CST) allows the researcher to specifically inhibit RCAS1 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis.

Quality Control

Oligonucleotide synthesis is monitored base by base through trityl analysis to ensure appropriate coupling efficiency. The oligo is subsequently purified by affinity-solid phase extraction. The annealed RNA duplex is further analyzed by mass spectrometry to verify the exact composition of the duplex. Each lot is compared to the previous lot by mass spectrometry to ensure maximum lot-to-lot consistency.

Directions for Use

CST recommends transfection with 100 nM RCAS1 siRNA I 48 to 72 hours prior to cell lysis. For transfection procedure, follow protocol provided by the transfection reagent manufacturer. Please feel free to contact CST with any questions on use.

Background

Receptor binding cancer antigen expressed on SiSo cells (RCAS1), also called estrogen receptor-binding fragment-associated gene 9 (EBAG9), originally identified as an estrogen-inducible gene (1) was recently found to play a novel role in the adaptive immune response by negatively regulating the cytolytic activity of cytotoxic T-lymphocytes (CTLs) (2). RCAS1 is conserved in phylogeny and is ubiquitously expressed in most human tissues and cells (3,4). There is evidence that tissue expression of RCAS1 is increased in a variety of malignancies, including cancers of the gastro-intestinal tract, liver, lung, breast, ovary, endometrium, and cervix. Moreover, levels of RCAS1 tissue expression are negatively correlated with the prognosis of patients harboring the aforementioned malignancies (4). It is also noteworthy that elevated levels of RCAS1 have been detected in the sera of cancer patients (4). Initial studies indicated that RCAS1 was secreted from cancer cells and functioned as a ligand for a putative receptor expressed on NK cells as well as T- and B-lymphocytes, inducing their apoptosis, which enabled cancer cells to evade immune surveillance (5,6). Subsequent studies have identified RCAS1 as a type-III transmembrane Golgi protein with the ability to regulate vesicle formation, secretion, and protein glycosylation (2,7-9). Indeed, it has been shown that RCAS1 overexpression negatively regulates the cytolytic function of CTLs by negatively regulating protein trafficking from the trans-Golgi to secretory lysosomes (2). Furthermore, RCAS1 overexpression delays vesicle transport from the ER to Golgi and causes components of the ER quality control and glycosylation machinery to mislocalize. As a consequence, RCAS1 induces the deposition of tumor-associated glycan antigens on the cell surface, which are thought to contribute to tumor pathogenesis through the mediation of adhesion, invasion, and metastasis (8,9).

1. Watanabe, T. et al. (1998) Mol Cell Biol 18, 442-9.
2. Rüder, C. et al. (2009) J Clin Invest 119, 2184-203.
3. Tsuchiya, F. et al. (2001) Biochem Biophys Res Commun 284, 2-10.
4. Giaginis, C. et al. (2009) Histol Histopathol 24, 761-76.
5. Matsushima, T. et al. (2001) Blood 98, 313-21.
6. Nakashima, M. et al. (1999) Nat Med 5, 938-42.
7. Reimer, T.A. et al. (2005) BMC Cancer 5, 47.
8. Wolf, J. et al. (2010) FASEB J 24, 4000-19.
9. Engelsberg, A. et al. (2003) J Biol Chem 278, 22998-3007.

注意事项：

Storage: RCAS1 siRNA I is supplied in RNAse-free water.
Aliquot and store at -20ºC.