pp-GalNAc-T13通过产生三聚体Tn抗原诱导鼠Lewis肺癌的高转移潜力。,Biochemical and Biophysical Research Communications

当前位置： X-MOL 学术 › Biochem. Biophys. Res. Commun. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

pp-GalNAc-T13通过产生三聚体Tn抗原诱导鼠Lewis肺癌的高转移潜力。
Biochemical and Biophysical Research Communications ( IF 2.5 ) Pub Date : 2012-02-07 , DOI: 10.1016/j.bbrc.2012.01.086
Yasuyuki Matsumoto ₁ , Qing Zhang , Kaoru Akita , Hiroshi Nakada , Kazunori Hamamura , Noriyo Tokuda , Akiko Tsuchida , Takeshi Matsubara , Tomoko Hori , Tetsuya Okajima , Keiko Furukawa , Takeshi Urano , Koichi Furukawa

Affiliation

为了分析癌症转移的机制，通过反复向小鼠Lewis肺癌子系H7和C4中注射获得高转移性亚系（H7-A，H7-Lu，H7-O，C4-sc和C4-ly） C57BL / 6小鼠。这些亚系在体外表现出增加的增殖和侵袭活性。神经节苷脂谱显示高转移亚系中的GM1表达低于亲本系。然后，我们通过稳定沉默H7细胞中的GM1合酶建立了GM1-Si-1和GM1-Si-2。这些GM1击倒克隆表现出增加的增殖和入侵。然后，我们探索了通过DNA微阵列在C4对C4 ly或H7对H7（GM1-Si）的组合中表达水平显着改变的基因。因此，pp-GalNAc-T13基因被鉴定为高转移亚系中的上调基因。pp-GalNAc-T13 cDNA稳定转染到C4（T13-TF）中导致入侵和运动能力增加。然后，使用各种抗体和凝集素进行免疫印迹和流式细胞仪。仅抗三聚体Tn抗体（mAb MLS128）在T13-TF克隆中显示三聚体Tn抗原的表达水平增加。此外，通过mAb MLS128进行了免疫沉淀/免疫印迹，从而鉴定了带有三聚体Tn抗原，即Syndecan-1的80 kDa条带。C4-sc（T13-KD）中内源性pp-GalNAc-T13的稳定沉默显示，皮下注射T13-KD克隆产生的原发性肿瘤与筋膜和腹膜的结合降低，并且肺转移明显低于对照克隆。这些数据表明pp-GalNAc-T13基因的高表达在Syndecan-1上产生了三聚体Tn抗原，

"点击查看英文标题和摘要"

pp-GalNAc-T13 induces high metastatic potential of murine Lewis lung cancer by generating trimeric Tn antigen.

In order to analyze the mechanisms for cancer metastasis, high metastatic sublines (H7-A, H7-Lu, H7-O, C4-sc, and C4-ly) were obtained by repeated injection of mouse Lewis lung cancer sublines H7 and C4 into C57BL/6 mice. These sublines exhibited increased proliferation and invasion activity in vitro. Ganglioside profiles exhibited lower expression of GM1 in high metastatic sublines than the parent lines. Then, we established GM1-Si-1 and GM1-Si-2 by stable silencing of GM1 synthase in H7 cells. These GM1-knockdown clones exhibited increased proliferation and invasion. Then, we explored genes that markedly altered in the expression levels by DNA microarray in the combination of C4 vs. C4-ly or H7 vs. H7 (GM1-Si). Consequently, pp-GalNAc-T13 gene was identified as up-regulated genes in the high metastatic sublines. Stable transfection of pp-GalNAc-T13 cDNA into C4 (T13-TF) resulted in increased invasion and motility. Then, immunoblotting and flow cytometry using various antibodies and lectins were performed. Only anti-trimeric Tn antibody (mAb MLS128), showed increased expression levels of trimeric Tn antigen in T13-TF clones. Moreover, immunoprecipitation/immunoblotting was performed by mAb MLS128, leading to the identification of an 80 kDa band carrying trimeric Tn antigen, i.e. Syndecan-1. Stable silencing of endogenous pp-GalNAc-T13 in C4-sc (T13-KD) revealed that primary tumors generated by subcutaneous injection of T13-KD clones showed lower coalescence to fascia and peritoneum, and significantly reduced lung metastasis than control clones. These data suggested that high expression of pp-GalNAc-T13 gene generated trimeric Tn antigen on Syndecan-1, leading to the enhanced metastasis.

更新日期：2019-11-01

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南