Zuber Chantal, Knackmuss Stefan, Zemora Georgeta, Reusch Uwe, Vlasova Ekaterina, Diehl Daniela, Mick Vera, Hoffmann Karin, Nikles Daphne, Fröhlich Thomas, Arnold Georg J, Brenig Bertram, Wolf Eckhard, Lahm Harald, Little Melvyn, Weiss Stefan
Laboratorium für Molekulare Biologie-Genzentrum-Institut für Biochemie der LMU München, Feodor-Lynen-Str. 25, D-81377 München, Germany.
J Mol Biol. 2008 May 2;378(3):530-9. doi: 10.1016/j.jmb.2008.02.004. Epub 2008 Feb 12.
The 37-kDa/67-kDa laminin receptor precursor/laminin receptor (LRP/LR) acting as a receptor for prions and viruses is overexpressed in various cancer cell lines, and their metastatic potential correlates with LRP/LR levels. We analyzed the tumorigenic fibrosarcoma cell line HT1080 regarding 37-kDa/67-kDa LRP/LR levels and its invasive potential. Compared to the less invasive embryonic fibroblast cell line NIH3T3, the tumorigenic HT1080 cells display approximately 1.6-fold higher cell-surface levels of LRP/LR. We show that anti-LRP/LR tools interfere with the invasive potential of HT1080 cells. Anti-LRP/LR single-chain variable fragment antibody (scFv) iS18 generated by chain shuffling from parental scFv S18 and its full-length version immunoglobulin G1-iS18 reduced the invasive potential of HT1080 cells significantly by 37% and 38%, respectively. HT1080 cells transfected with lentiviral plasmids expressing small interfering RNAs directed against LRP mRNA showed reduced LRP levels by approximately 44%, concomitant with a significant decrease in the invasive potential by approximately 37%. The polysulfated glycans HM2602 and pentosan polysulfate (SP-54), both capable of blocking LRP/LR, reduced the invasive potential by 20% and 35%, respectively. Adhesion of HT1080 cells to laminin-1 was significantly impeded by scFv iS18 and immunoglobulin G1-iS18 by 60% and 68%, respectively, and by SP-54 and HM2602 by 80%, suggesting that the reduced invasive capacity achieved by these tools is due to the perturbation of the LRP/LR-laminin interaction on the cell surface. Our in vitro data suggest that reagents directed against LRP/LR or LRP mRNA such as antibodies, polysulfated glycans, or small interfering RNAs, previously shown to encompass an anti-prion activity by blocking or downregulating the prion receptor LRP/LR, might also be potential cancer therapeutics blocking metastasis by interfering with the LRP/LR-laminin interaction in neoplastic tissues.
作为朊病毒和病毒受体的37 kDa/67 kDa层粘连蛋白受体前体/层粘连蛋白受体（LRP/LR）在多种癌细胞系中过表达，其转移潜能与LRP/LR水平相关。我们分析了致瘤性纤维肉瘤细胞系HT1080的37 kDa/67 kDa LRP/LR水平及其侵袭潜能。与侵袭性较低的胚胎成纤维细胞系NIH3T3相比，致瘤性HT1080细胞的LRP/LR细胞表面水平高约1.6倍。我们发现抗LRP/LR工具可干扰HT1080细胞的侵袭潜能。通过从亲本单链可变片段抗体（scFv）S18进行链改组产生的抗LRP/LR单链可变片段抗体iS18及其全长版本免疫球蛋白G1-iS18分别使HT1080细胞的侵袭潜能显著降低了37%和38%。用表达针对LRP mRNA的小干扰RNA的慢病毒质粒转染的HT1080细胞，其LRP水平降低了约44%，同时侵袭潜能显著降低了约37%。两种能够阻断LRP/LR的多硫酸化聚糖HM2602和戊聚糖多硫酸盐（SP-54）分别使侵袭潜能降低了20%和35%。scFv iS18和免疫球蛋白G1-iS18分别使HT1080细胞与层粘连蛋白-1的黏附显著受阻60%和68%，SP-54和HM2602使其受阻80%，这表明这些工具实现的侵袭能力降低是由于细胞表面LRP/LR-层粘连蛋白相互作用受到干扰。我们的体外数据表明，针对LRP/LR或LRP mRNA的试剂，如抗体、多硫酸化聚糖或小干扰RNA，先前已证明通过阻断或下调朊病毒受体LRP/LR具有抗朊病毒活性，它们也可能是通过干扰肿瘤组织中LRP/LR-层粘连蛋白相互作用来阻断转移的潜在癌症治疗药物。
