Shin Seung-Shick, Namkoong Jin, Wall Brian A, Gleason Ryan, Lee Hwa Jin, Chen Suzie
Department of Chemical Biology, Ernest Mario School of Pharmacy, Susan Lehman Cullman Laboratory for Cancer Research, Rutgers University, Piscataway, NJ, USA.
Pigment Cell Melanoma Res. 2008 Jun;21(3):368-78. doi: 10.1111/j.1755-148X.2008.00452.x. Epub 2008 Apr 23.
Previously, we reported a transgenic mouse line, TG-3, that develops spontaneous melanoma with 100% penetrance. We demonstrated that ectopic expression of Grm1 in melanocytes was sufficient to induce melanoma in vivo. In this present study, the transforming properties of Grm1 in two cultured immortalized melanocytes were investigated. We showed that, in contrast to parental melanocytes, these Grm1-clones have lost their requirement of TPA supplement for proliferation and have acquired the ability to form colonies in semi-solid medium. Xenografts of these cells formed robust tumors in both immunodeficient nude and syngeneic mice with a short latency (3-5 days). The malignancy of these cells was demonstrated by angiogenesis and invasion to the muscle and the intestine. The requirement of Grm1 expression for the maintenance of transformation was demonstrated by an inducible siRNA system. Induction of expression of siRNA for Grm1 reduced the number of proliferating/viable cells in vitro and suppressed in vivo xenografted tumor growth in comparison with control. Taken together, these results showed that expression of exogeneously introduced Grm1 is sufficient to induce full transformation of immortalized melanocytes.
此前,我们报道了一种转基因小鼠品系TG-3,其会100%自发发生黑色素瘤。我们证明,黑素细胞中Grm1的异位表达足以在体内诱导黑色素瘤。在本研究中,我们研究了Grm1在两种永生化黑素细胞系中的转化特性。我们发现,与亲代黑素细胞相比,这些Grm1克隆在增殖时不再需要添加佛波酯(TPA),并且获得了在半固体培养基中形成集落的能力。这些细胞的异种移植在免疫缺陷的裸鼠和同基因小鼠中均能迅速形成肿瘤,潜伏期较短(3-5天)。这些细胞的恶性特征表现为血管生成以及侵袭肌肉和肠道。通过诱导型siRNA系统证明了维持转化需要Grm1的表达。与对照相比,诱导表达针对Grm1的siRNA可减少体外增殖/存活细胞的数量,并抑制体内异种移植肿瘤的生长。综上所述,这些结果表明,外源导入的Grm1的表达足以诱导永生化黑素细胞的完全转化。