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腺病毒介导白细胞介素12基因转导的人脐带间充质干细胞在体内外均抑制卵巢癌细胞生长

[Human umbilical cord mesenchymal stem cells with adenovirus-mediated interleukin 12 gene transduction inhibits the growth of ovarian carcinoma cells both in vitro and in vivo].

作者信息

Zhao Wen-hong, Cheng Jian-xin, Shi Peng-fei, Huang Jun-yan

机构信息

Department of Gynecology and Obstetrics, Fourth Hospital of Hebei Medical University, Shijiazhuang, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2011 May;31(5):903-7.

PMID:21602155
Abstract

OBJECTIVE

To study the inhibitory effect of human umbilical cord mesenchymal stem cells (UC-MSCs) infected by a adenoviral vector containing interleukin 12 (IL-12) gene on the proliferation of ovarian carcinoma SKOV3 in vitro and the growth of tumor explants in nude mice.

METHODS

Cultured human UC-MSCs were infected with the recombinant adenovirus vector harboring IL-12 gene to establish the IL-12-expressing cell line AdIL-12-MSCs. Western blotting and RT-PCR were used to detect IL-12 expressions in AdIL-12-MSCs at the protein and mRNA levels, respectively. ELISA were used to detect IL-12 content in the supernatant of AdIL-12-MSCs, whose effect on the proliferation and apoptosis of ovarian carcinoma SKOV3 cells was evaluated with MTT assay and flow cytometry, respectively. In a nude mouse model bearing subcutaneous SKOV3 tumor explants, AdIL-12-MSCs were infused via the tail vein and the inhibitory effect on the tumor growth was observed.

RESULTS

The exogenous IL-12 gene was successfully transduced into UC-MSCs by the recombinant adenovirus vector, resulting in efficient IL-12 expression in the cell at both the protein and mRNA levels. The supernatant of AdIL-12-MSCs significantly inhibited the proliferation of SKOV3 cells and induced cellular apoptosis in vitro as compared with UC-MSC supernatant. In the tumor-bearing nude mouse model, the transplantation of AdIL-12-MSCs significantly inhibited the growth of SKOV3 tumor explants (P<0.05).

CONCLUSION

Human UC-MSCs with IL-12 gene transduction, which express IL-12 at protein and mRNA levels, can inhibit the proliferation and induce apoptosis of ovarian carcinoma SKOV3 cells in vitro, and suppress the growth of ovarian cancer explants in nude mice.

摘要

目的

研究携带白细胞介素12(IL-12)基因的腺病毒载体感染人脐带间充质干细胞(UC-MSCs)后对卵巢癌SKOV3细胞体外增殖及裸鼠体内肿瘤组织生长的抑制作用。

方法

用携带IL-12基因的重组腺病毒载体感染培养的人UC-MSCs,建立表达IL-12的细胞系AdIL-12-MSCs。分别采用蛋白质印迹法和逆转录-聚合酶链反应(RT-PCR)从蛋白质和mRNA水平检测AdIL-12-MSCs中IL-12的表达。采用酶联免疫吸附测定(ELISA)检测AdIL-12-MSCs培养上清中IL-12的含量,分别用噻唑蓝(MTT)比色法和流式细胞术评估其对卵巢癌SKOV3细胞增殖和凋亡的影响。在皮下接种SKOV3肿瘤组织的裸鼠模型中,经尾静脉注入AdIL-12-MSCs,观察其对肿瘤生长的抑制作用。

结果

重组腺病毒载体成功将外源性IL-12基因转导入UC-MSCs,使细胞在蛋白质和mRNA水平均高效表达IL-12。与UC-MSCs培养上清相比,AdIL-12-MSCs培养上清在体外显著抑制SKOV3细胞增殖并诱导细胞凋亡。在荷瘤裸鼠模型中,移植AdIL-12-MSCs显著抑制SKOV3肿瘤组织生长(P<0.05)。

结论

转导IL-12基因的人UC-MSCs在蛋白质和mRNA水平均表达IL-12,可在体外抑制卵巢癌SKOV3细胞增殖并诱导其凋亡,在裸鼠体内可抑制卵巢癌组织生长。

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