Kakinuma Hideaki, Bergert Elizabeth R, Spitzweg Christine, Cheville John C, Lieber Michael M, Morris John C
Department of Endocrinology, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA.
Cancer Res. 2003 Nov 15;63(22):7840-4.
Prostate cancer is one of the most promising candidates for sodium iodide symporter (NIS)-mediated gene therapy. Adenovirus-mediated expression of NIS that is driven by prostate-specific promoters induces generous radioiodine accumulation in prostate cancer cells that may be used for therapy with (131)I. We have recently developed a replication-deficient adenovirus carrying the human NIS cDNA linked to a composite probasin promoter, ARR(2)PB, aiming toward specific expression of the human NIS gene (h-NIS) in prostate tissue for targeted radioactive iodide therapy of prostate cancer (Ad-ARR(2)PB/hNIS). The ability of Ad-ARR(2)PB/hNIS to cause NIS expression in tumor cells was characterized by iodide uptake assay and compared with Ad-CMV/hNIS in which the h-NIS expression is driven by the cytomegalovirus (CMV) promoter. Androgen-dependent prostate cancer cell lines (LNCaP) and non-prostate origin tumor cell lines (SNU449, MCF-7, HCT116, OVCAR-3, and Panc-1) were infected with the viral constructs, and perchlorate-sensitive (125)I uptake and NIS protein expression were measured. Ad-ARR(2)PB/hNIS-infected LNCaP cells showed androgen-dependent and perchlorate-sensitive iodide uptake. Iodide accumulation in LNCaP cells infected with Ad-ARR(2)PB/hNIS, followed by incubation with synthetic androgen, was 5.3-fold increased compared with those coincubated with perchlorate (15,184 +/- 1,173 cpm versus 2,837 +/- 187 cpm). Ad-ARR(2)PB/hNIS-infected LNCaP cells revealed a 3.2-fold increase of iodide accumulation compared with those infected with Ad-CMV/hNIS (multiplicity of infection = 30). Iodide uptake in a panel of non-prostate tumor cell lines infected with Ad-ARR(2)PB/hNIS was no more than 2,500 cpm, demonstrating the tissue specificity of this construct. These results indicate that Ad-ARR(2)PB/hNIS can be used to achieve high-magnitude and tissue-specific expression of h-NIS in prostate tissue and is a promising candidate for cancer gene therapy of prostate cancer.
前列腺癌是碘化钠同向转运体（NIS）介导的基因治疗最有前景的候选对象之一。由前列腺特异性启动子驱动的腺病毒介导的NIS表达可诱导前列腺癌细胞大量积累放射性碘，这可用于（131）I治疗。我们最近开发了一种复制缺陷型腺病毒，其携带与人NIS cDNA相连的复合前列腺素启动子ARR(2)PB，旨在使人NIS基因（h-NIS）在前列腺组织中特异性表达，用于前列腺癌的靶向放射性碘治疗（Ad-ARR(2)PB/hNIS）。通过碘摄取试验对Ad-ARR(2)PB/hNIS在肿瘤细胞中引起NIS表达的能力进行了表征，并与h-NIS表达由巨细胞病毒（CMV）启动子驱动的Ad-CMV/hNIS进行了比较。用病毒构建体感染雄激素依赖性前列腺癌细胞系（LNCaP）和非前列腺来源的肿瘤细胞系（SNU449、MCF-7、HCT116、OVCAR-3和Panc-1），并测量高氯酸盐敏感性（125）I摄取和NIS蛋白表达。Ad-ARR(2)PB/hNIS感染的LNCaP细胞表现出雄激素依赖性和高氯酸盐敏感性碘摄取。与用高氯酸盐共同孵育的细胞相比，Ad-ARR(2)PB/hNIS感染的LNCaP细胞在用合成雄激素孵育后碘积累增加了5.3倍（15,184 +/- 1,173 cpm对2,837 +/- 187 cpm）。与Ad-CMV/hNIS感染的细胞相比（感染复数=30），Ad-ARR(2)PB/hNIS感染的LNCaP细胞的碘积累增加了3.2倍。用Ad-ARR(2)PB/hNIS感染的一组非前列腺肿瘤细胞系中的碘摄取不超过2,500 cpm，证明了该构建体的组织特异性。这些结果表明，Ad-ARR(2)PB/hNIS可用于在前列腺组织中实现h-NIS的高表达和组织特异性表达，是前列腺癌基因治疗的一个有前景的候选对象。
