[携带小鼠β-防御素2的间充质干细胞对小鼠恶性腹水的抑制作用]

[Inhibitory effect of mesenchymal stem cells carrying murine beta defensin 2 on malignant ascites in mice].

作者信息

Wang Guo-Qing, Xu Jian-Rong, Wang Rui, Li Hong-Xia, Xu Ning, Du Xiao-Bo, Wei Yu-Quan

机构信息

State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan, 610041, P. R. China.

出版信息

Ai Zheng. 2006 Jun;25(6):657-62.

PMID:16764757

Abstract

BACKGROUND & OBJECTIVE: Murine beta defensin 2 (MBD2) is a small antimicrobial peptide of the innate immune system. It provides a critical link between the innate immune system and the adaptive immune response. This study was to develop a set of MBD2-lentivirus system, and observe its inhibitory effect on malignant ascites in mice.

METHODS

MBD2 RNA was extracted from kidneys of BALB/c mice, and the fragment of MBD2 was amplified by reverse transcription-polymerase chain reaction (RT-PCR). MBD2 plasmid was constructed, and the lentivirus system expressing MBD2 was produced in 293FT cells. Mesenchymal stem cells (MSCs) were then infected with MBD2-Lentivirus, and stably infected cells (MBD2-MSCs) were selected. The expression of MBD2 was detected by RT-PCR. The biological function of MBD2 was evaluated by dendritic cell (DC) migration experiment. BALB/c mice bearing intraperitoneal MethA tumors were injected intraperitoneally with MBD2-MSCs at the 2nd, 4th, 6th, and 8th day after inoculation. Ascites status and survival status of the mice were observed. Long-term survivors were inoculated intraperitoneally with MethA tumor cells again to observe their survival status.

RESULTS

The expression of MBD2 in MBD2-MSCs was verified by RT-PCR. Numbers of migrated DCs were significantly increased in MBD2-MSC group than in normal saline (NS) group, MSC group, and null lentivirus (Null) group (43+/-8 vs. 8+/-1, 14+/-2, and 12+/-3, P<0.01). The volume of tumor ascites was significantly less in MBD2-MSC group than in NS group, MSC group, and Null group [(3.0+/-1.0) ml vs. (10.8+/-1.0) ml, (10.2+/-1.3) ml, and (9.8+/-1.6) ml, P<0.05]. Some mice in MBD2 group were still alive 40 days after tumor inoculation, while none in control groups survived up to 20 days after tumor inoculation (P<0.05). After inoculated with MethA tumor cells again, the mice in MBD2-MSC group still survived longer than those in control groups.

CONCLUSION

MBD2-lentivirus gene therapy system is constructed successfully, and can inhibit the formation of malignant ascites.

摘要

背景与目的

小鼠β-防御素2（MBD2）是一种先天性免疫系统的小抗菌肽。它在先天性免疫系统和适应性免疫反应之间提供了关键联系。本研究旨在构建一套MBD2慢病毒系统，并观察其对小鼠恶性腹水的抑制作用。

方法

从BALB/c小鼠肾脏中提取MBD2 RNA，通过逆转录-聚合酶链反应（RT-PCR）扩增MBD2片段。构建MBD2质粒，并在293FT细胞中生产表达MBD2的慢病毒系统。然后用MBD2慢病毒感染间充质干细胞（MSC），筛选出稳定感染的细胞（MBD2-MSC）。通过RT-PCR检测MBD2的表达。通过树突状细胞（DC）迁移实验评估MBD2的生物学功能。接种腹腔内MethA肿瘤的BALB/c小鼠在接种后第2、4、6和8天腹腔注射MBD2-MSC。观察小鼠的腹水状态和生存状态。对长期存活者再次腹腔接种MethA肿瘤细胞，观察其生存状态。

结果

通过RT-PCR验证了MBD2-MSC中MBD2的表达。MBD2-MSC组迁移的DC数量显著多于生理盐水（NS）组、MSC组和空慢病毒（Null）组（43±8对8±1、14±2和12±3，P<0.01）。MBD2-MSC组的肿瘤腹水体积显著小于NS组、MSC组和Null组[（3.0±1.0）ml对（10.8±1.0）ml、（10.2±1.3）ml和（9.8±1.6）ml，P<0.05]。MBD2组的一些小鼠在肿瘤接种后40天仍存活，而对照组中没有一只小鼠在肿瘤接种后存活至20天（P<0.05）。再次接种MethA肿瘤细胞后，MBD2-MSC组的小鼠存活时间仍长于对照组。