利用 PSA 荧光素酶报告质粒在小鼠模型中监测前列腺癌的生长和转移。

Monitoring of prostate cancer growth and metastasis using a PSA luciferase report plasmid in a mouse model.

机构信息

Department of Urology, the First Affiliated Hospital of Zhejiang University Medical College, Hangzhou, 310003, China.

出版信息

Asian Pac J Trop Med. 2014 Nov;7(11):879-83. doi: 10.1016/S1995-7645(14)60153-X. Epub 2014 Oct 22.

DOI:10.1016/S1995-7645(14)60153-X

PMID:25441987

Abstract

OBJECTIVE

To construct a PSA luciferase report plasmid and monitor the growth and metastasis of prostate cancer after emasculation in SCID mice.

METHODS

PSA promoter sequence and luciferase gene were amplified by PCR and subsequently inserted into pZsGreen1-1 vector to construct pPSA-FL-Luc vector. LNCaP cells that were stably transfected with pPSA-FL-Luc were used to establish a SCID mouse xenograft model. Then, the growth and metastasis of prostate cancer were monitored via living imaging.

RESULTS

We successfully constructed a PSA luciferase plasmid, pPSA-FL-Luc. DHT enhanced luciferase activity in a concentration-dependent manner in 293T cells with pPSA-FL-Luc transfection. Prostate cancer SCID mouse model was established with pPSA-FL-Luc transfected LNCaP cells. In tumor bearing mice with or without emasculation, pPSA-FL-Luc plasmid was applied to monitored tumor growth and metastasis based on bioluminescence imaging.

CONCLUSIONS

We construct a pPSA-FL-Luc plasmid, which stably expresses luciferase and can be applied to monitor tumor development in a prostate SCID mouse model.

摘要

目的

构建 PSA 荧光素酶报告质粒，并监测去势后 SCID 小鼠前列腺癌的生长和转移。

方法

通过 PCR 扩增 PSA 启动子序列和荧光素酶基因，然后将其插入 pZsGreen1-1 载体中，构建 pPSA-FL-Luc 载体。用 pPSA-FL-Luc 稳定转染的 LNCaP 细胞建立 SCID 小鼠异种移植模型。然后通过活体成像监测前列腺癌的生长和转移。

结果

我们成功构建了 PSA 荧光素酶质粒 pPSA-FL-Luc。在转染了 pPSA-FL-Luc 的 293T 细胞中，DHT 以浓度依赖的方式增强了荧光素酶的活性。用 pPSA-FL-Luc 转染的 LNCaP 细胞建立了前列腺癌 SCID 小鼠模型。在去势或不去势的荷瘤小鼠中，应用 pPSA-FL-Luc 质粒通过生物发光成像监测肿瘤生长和转移。