Department of Biotechnology, The Catholic University of Korea, Bucheon 420-743, Republic of Korea.
Int J Oncol. 2011 Oct;39(4):853-61. doi: 10.3892/ijo.2011.1095. Epub 2011 Jun 22.
Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world. However, little is known regarding the molecular mechanism of HCC development and progression and effective therapeutic methods. Recently, the granulin-epithelin precursor (GEP) was reported as a novel growth factor that can control HCC cell proliferation. Using the CAPSID program, we designed three small interfering RNAs (siRNAs) targeting the GEP gene (GEP-siRNA1, 2 and 3) and examined their tumor regression and suppression effects on cell proliferation. GEP-siRNA1 exhibited the strongest anti-proliferative effect among the GEP-siRNAs, in a time-dependent manner. To increase the biostability of the siRNA, we also constructed a short hairpin RNA (shRNA) using an H1/TO promoter with the same sequence of GEP-siRNA1 (GEP-shRNA). GEP-shRNA decreased the expression levels of GEP and tumor cell growth via cell cycle arrest at the G2/M stage and down-regulation of the cell proliferation proteins cyclin D1 and α-tubulin. Furthermore, GEP-shRNA inhibited tumor growth significantly after intratumoral injection into tumor-bearing Balb/C nude mice. Taken together, these results represent the first therapeutic application of RNA interference to GEP, which is a promising target molecule for HCC treatment, as an approach for the suppression of HCC cell proliferation.
肝细胞癌（HCC）是世界上第五种最常见的癌症。然而，人们对 HCC 的发展和进展的分子机制以及有效的治疗方法知之甚少。最近，颗粒蛋白-上皮素前体（GEP）被报道为一种新的生长因子，可以控制 HCC 细胞增殖。使用 CAPSID 程序，我们设计了针对 GEP 基因的三种小干扰 RNA（siRNA）（GEP-siRNA1、2 和 3），并检查了它们对肿瘤消退和细胞增殖抑制的作用。在时间依赖性方式下，GEP-siRNA1 在 GEP-siRNA 中表现出最强的抗增殖作用。为了提高 siRNA 的生物稳定性，我们还使用具有与 GEP-siRNA1 相同序列的 H1/TO 启动子构建了短发夹 RNA（shRNA）（GEP-shRNA）。GEP-shRNA 通过细胞周期阻滞在 G2/M 期和下调细胞增殖蛋白 cyclin D1 和α-微管蛋白来降低 GEP 和肿瘤细胞生长的表达水平。此外，GEP-shRNA 经肿瘤内注射到荷瘤 Balb/C 裸鼠后显著抑制肿瘤生长。总之，这些结果代表了 RNA 干扰在 GEP 治疗中的首次应用，作为抑制 HCC 细胞增殖的方法，GEP 是 HCC 治疗的有前途的靶分子。
