Takis srl, Rome, Italy.
J Cell Physiol. 2013 Jan;228(1):58-64. doi: 10.1002/jcp.24103.
The kinesin spindle protein (KSP), a member of the kinesin superfamily of microtubule-based motors, plays a critical role in mitosis as it mediates centrosome separation and bipolar spindle assembly and maintenance. Inhibition of KSP function leads to cell cycle arrest at mitosis with the formation of monoastral microtubule arrays, and ultimately, to cell death. Several KSP inhibitors are currently being studied in clinical trials and provide new opportunities for the development of novel anticancer therapeutics. RNA interference (RNAi) may represent a powerful strategy to interfere with key molecular pathways involved in cancer. In this study, we have established an efficient method for intratumoral delivery of siRNA. We evaluated short interfering RNA (siRNA) duplexes targeting luciferase as surrogate marker or KSP sequence. To examine the potential feasibility of RNAi therapy, the siRNA was transfected into pre-established lesions by means of intratumor electro-transfer of RNA therapeutics (IERT). This technology allowed cell permeation of the nucleic acids and to efficiently knock down gene expression, albeit transiently. The KSP-specific siRNA drastically reduced outgrowth of subcutaneous melanoma and ovarian cancer lesions. Our results show that intratumoral electro-transfer of siRNA is feasible and KSP-specific siRNA may provide a novel strategy for therapeutic intervention.
驱动蛋白纺锤体蛋白（KSP）是微管动力蛋白超家族的成员，在有丝分裂中起着关键作用，因为它介导中心体分离和双极纺锤体的组装和维持。KSP 功能的抑制会导致有丝分裂细胞周期停滞，形成单星微管阵列，并最终导致细胞死亡。目前正在临床试验中研究几种 KSP 抑制剂，为开发新型抗癌治疗方法提供了新的机会。RNA 干扰（RNAi）可能代表了一种干扰癌症相关关键分子途径的有效策略。在这项研究中，我们建立了一种有效的肿瘤内递送 siRNA 的方法。我们评估了针对荧光素酶的短干扰 RNA（siRNA）双链作为替代标志物或 KSP 序列。为了检查 RNAi 治疗的潜在可行性，通过肿瘤内 RNA 治疗电转移（IERT）将 siRNA 转染到预先建立的病变中。该技术允许核酸细胞渗透，并有效地敲低基因表达，尽管是短暂的。KSP 特异性 siRNA 大大减少了皮下黑素瘤和卵巢癌病变的生长。我们的结果表明，肿瘤内电转移 siRNA 是可行的，并且 KSP 特异性 siRNA 可能为治疗干预提供一种新策略。
