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NPM1 在耐药性膀胱癌侵袭和迁移中的作用。

The Role of NPM1 in the Invasion and Migration of Drug Resistant Bladder Cancer.

机构信息

Clinical Laboratory Medicine, Peking University Ninth School of Clinical Medicine, Beijing, 100038, China.

Clinical Laboratory Medicine, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, China .

出版信息

Urol J. 2021 Jul 6;18(4):452-459. doi: 10.22037/uj.v18i.6087.

Abstract

PURPOSE

To study the difference of tumor progression caused by differential expression of NPM1 in drug-resistant bladder cancer.

MATERIALS AND METHODS

The expression of NPM1 was analyzed by PCR and Western blot. NPM1 silencing bladder cancer cells (T24/DDP Lv-NPM1, PUMC-91/DDP Lv-NPM1) and overexpressing bladder cancer cells (T24/ DDP Lv5-NPM1, PUMC-91/DDP Lv5-NPM1) were established by lentivirus and limited dilution method. The efficiency of gene interference was detected by fluorescence microscopy and Western blot. The migration ability and invasion ability of tumor in vitro were analyzed by wound healing assay and transwell cell invasion test, and the tumorigenic ability in vivo was judged by nude mouse tumorigenicity assay.

RESULTS

Compared with the corresponding negative control group, both NPM1 silencing cell lines T24/DDP Lv-NPM1 and PUMC-91/DDP Lv-NPM1 showed strong migration ability and high invasive ability. At the same time, there was no significant difference in migration ability and the invasive cells proportion between NPM1 overexpressing cell line and related negative control group. NPM1 silencing bladder cancer cells had obvious tumorigenicity in vivo.

CONCLUSION

NPM1 silencing cells had significant migration and invasion ability. The silencing of NPM1 will accelerate tumorigenicity of drug resistant bladder cancer. Differential expression of NPM1 is of great value in monitoring the progression of drug-resistant bladder cancer.

摘要

目的

研究 NPM1 差异表达导致耐药膀胱癌肿瘤进展的差异。

材料与方法

采用 PCR 和 Western blot 分析 NPM1 的表达。通过慢病毒和有限稀释法建立 NPM1 沉默膀胱癌细胞(T24/DDP Lv-NPM1、PUMC-91/DDP Lv-NPM1)和过表达膀胱癌细胞(T24/ DDP Lv5-NPM1、PUMC-91/DDP Lv5-NPM1)。荧光显微镜和 Western blot 检测基因干扰效率。划痕愈合试验和 Transwell 细胞侵袭试验分析肿瘤体外迁移和侵袭能力,裸鼠成瘤试验判断体内致瘤能力。

结果

与相应的阴性对照组相比,NPM1 沉默细胞系 T24/DDP Lv-NPM1 和 PUMC-91/DDP Lv-NPM1 均表现出较强的迁移能力和高侵袭能力。同时,NPM1 过表达细胞系与相关阴性对照组之间的迁移能力和侵袭细胞比例无显著差异。NPM1 沉默膀胱癌细胞在体内具有明显的致瘤性。

结论

NPM1 沉默细胞具有明显的迁移和侵袭能力。NPM1 的沉默会加速耐药膀胱癌的肿瘤发生。NPM1 的差异表达对监测耐药膀胱癌的进展具有重要价值。

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