Functional roles of Src and Fgr in ovarian carcinoma.
Functional roles of Src and Fgr in ovarian carcinoma.
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DOI：
10.1158/1078-0432.ccr-10-2081
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发表时间：
2011-04-01
期刊：
影响因子：
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通讯作者：
Sood AK
中科院分区：
文献类型：
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作者：
Kim HS;Han HD;Armaiz-Pena GN;Stone RL;Nam EJ;Lee JW;Shahzad MM;Nick AM;Lee SJ;Roh JW;Nishimura M;Mangala LS;Bottsford-Miller J;Gallick GE;Lopez-Berestein G;Sood AK
Src是一个有吸引力的靶点，因为它在许多恶性肿瘤中过表达，包括卵巢癌。然而，Src沉默对其他Src家族激酶（SFK）的影响尚不清楚。我们假设其他SFK成员可以弥补Src活性的缺乏。在卵巢癌细胞系中，通过MTT测定来检查Src或Fgr沉默后的细胞活力。实时荧光定量RT-PCR检测Src基因沉默后卵巢癌细胞中SFKs的表达。使用掺入到壳聚糖纳米颗粒中的siRNA（siRNA/CH-NP）检查体内Src和/或Fgr沉默的治疗效果。采用免疫组化法检测卵巢肿瘤组织中的微血管密度（MVD）、细胞增殖和凋亡标志物。Src沉默增强多西他赛在SKOV 3 ip 1和HeyA 8细胞中的细胞毒性。此外，与对照siRNA/CH-NP相比，使用siRNA/CH-NP与多西他赛组合的Src沉默导致肿瘤生长的显著抑制（SKOV 3 ip 1中减少81.8%，P = 0.017; HeyA 8中减少84.3%，P < 0.005）。这些作用通过降低肿瘤细胞增殖和血管生成以及增加肿瘤细胞凋亡来介导。接下来，我们评估了Src沉默对卵巢癌细胞系中其他SFK成员的影响。Src沉默导致Fgr水平显著增加。双Src和Fgr沉默在体外导致细胞凋亡的增加，介导的半胱天冬酶和AKT活性增加。此外，在HeyA 8模型中，与单独沉默Src或Fgr相比，使用siRNA/CH-NP在体内双重沉默Src和Fgr导致肿瘤生长的最大降低（68.8%，P < 0.05）。这项研究表明，除了Src，Fgr在卵巢癌生长中起着重要的生物学作用，可能是一个重要的靶点。
Src is an attractive target because it is overexpressed in a number of malignancies, including ovarian cancer. However, the effect of Src silencing on other Src family kinases (SFKs) is not known. We hypothesized that other SFK members could compensate for the lack of Src activity. Cell viability following either Src or Fgr silencing was examined in ovarian cancer cell lines by MTT assay. Expression of SFKs after Src silencing in ovarian cancer cells was examined by real-time RT-PCR. Therapeutic effect of in vivo Src and/or Fgr silencing was examined using siRNA incorporated into chitosan nanoparticles (siRNA/CH-NP). Microvessel density (MVD), cell proliferation, and apoptosis markers were determined by immunohistochemical staining in ovarian tumor tissues. Src silencing enhanced cytotoxicity of docetaxel in both SKOV3ip1 and HeyA8 cells. In addition, Src silencing using siRNA/CH-NP in combination with docetaxel resulted in significant inhibition of tumor growth compared to control siRNA/CH-NP (81.8% reduction in SKOV3ip1, P = 0.017; 84.3% reduction in HeyA8, P < 0.005). These effects were mediated by decreased tumor cell proliferation and angiogenesis, and increased tumor cell apoptosis. Next, we assessed the effects of Src silencing on other SFK members in ovarian cancer cell lines. Src silencing resulted in significantly increased Fgr levels. Dual Src and Fgr silencing in vitro resulted in increased apoptosis that was mediated by increased caspase and AKT activity. In addition, dual silencing of Src and Fgr in vivo using siRNA/CH-NP resulted in the greatest reduction in tumor growth compared to silencing of either Src or Fgr alone in the HeyA8 model (68.8%, P < 0.05). This study demonstrates that, in addition to Src, Fgr plays a biologically significant role in ovarian cancer growth and might represent an important target.