Zhao Wen-hong, Cheng Jian-xin, Shi Peng-fei, Huang Jun-yan
Department of Gynecology and Obstetrics, Fourth Hospital of Hebei Medical University, Shijiazhuang, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2011 May;31(5):903-7.
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.
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.
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).
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.
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。
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。
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)。
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。