Mesenchymal stem cells are efficiently transduced with adenoviruses bearing type 35-derived fibers and the transduced cells with the IL-28A gene produces cytotoxicity to lung carcinoma cells co-cultured.
Mesenchymal stem cells are efficiently transduced with adenoviruses bearing type 35-derived fibers and the transduced cells with the IL-28A gene produces cytotoxicity to lung carcinoma cells co-cultured.
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DOI：
10.1186/1471-2407-14-713
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发表时间：
2014-09-25
期刊：
影响因子：
3.8
通讯作者：
Tagawa M
中科院分区：
文献类型：
--
作者：
Suzuki T;Kawamura K;Li Q;Okamoto S;Tada Y;Tatsumi K;Shimada H;Hiroshima K;Yamaguchi N;Tagawa M
5型腺病毒（Ad5）转染人间充质干细胞（MSCs）的效果有限，因为柯萨奇腺病毒受体（CAR）分子的表达水平较低。我们研究了用35型Ad取代5型Ad的纤维区可能改善Ad介导的基因转移。用流式细胞术检测5型和35型Ad的主要受体CAR和CD46分子的表达水平。我们构建了Ad5或Ad5修饰的35型纤维区（AdF35）绿色荧光蛋白基因表达载体，并用流式细胞术检测其对MSCs的感染性。我们用比色法研究了白细胞介素-28A基因转导的MSCs对人肺癌细胞的抗肿瘤作用。用聚合酶链反应检测IL-28A受体的表达。荧光素酶法检测MSCs转录调控区启动子活性，裸鼠共注射人肿瘤细胞检测MSCs促肿瘤生长能力。MSCs表达CD46分子，但很少表达CAR分子，随后被AdF35转导，而不被Ad5转导。转染IL-28A基因的间充质干细胞的生长与未转染的细胞相同，因为间充质干细胞对IL-28A受体呈阴性。然而，il - 28a转导的MSCs抑制了共培养的肺癌细胞的生长，而表达β-半乳糖苷酶基因的AdF35转导的MSCs则没有抑制作用。环合酶-2基因的一个调控区域具有比其他肿瘤启动子更高的转录活性，但比巨细胞病毒启动子低，并且MSCs本身在体内不支持肿瘤生长。AdF35是一种适合转导对ad5介导的基因转移具有抗性的间充质干细胞的载体。通过巨细胞病毒启动子激活外源基因的AdF35感染的间充质干细胞可以作为载体将基因产物传递到靶细胞。
Transduction of human mesenchymal stem cells (MSCs) with type 5 adenoviruses (Ad5) is limited in the efficacy because of the poor expression level of the coxsackie adenovirus receptor (CAR) molecules. We examined a possible improvement of Ad-mediated gene transfer in MSCs by substituting the fiber region of type 5 Ad with that of type 35 Ad. Expression levels of CAR and CD46 molecules, which are the major receptors for type 5 and type 35 Ad, respectively, were assayed with flow cytometry. We constructed vectors expressing the green fluorescent protein gene with Ad5 or modified Ad5 bearing the type 35 fiber region (AdF35), and examined the infectivity to MSCs with flow cytometry. We investigated anti-tumor effects of MSCs transduced with interleukin (IL)-28A gene on human lung carcinoma cells with a colorimetric assay. Expression of IL-28A receptors was tested with the polymerase chain reaction. A promoter activity of transcriptional regulatory regions in MSCs was determined with a luciferase assay and a tumor growth-promoting ability of MSCs was tested with co-injection of human tumor cells in nude mice. MSCs expressed CD46 but scarcely CAR molecules, and subsequently were transduced with AdF35 but not with Ad5. Growth of MSCs transduced with the IL-28A gene remained the same as that of untransduced cells since MSCs were negative for the IL-28A receptors. The IL-28A-transduced MSCs however suppressed growth of lung carcinoma cells co-cultured, whereas MSCs transduced with AdF35 expressing the β-galactosidase gene did not. A regulatory region of the cyclooygenase-2 gene possessed transcriptional activities greater than other tumor promoters but less than the cytomegalovirus promoter, and MSCs themselves did not support tumor growth in vivo. AdF35 is a suitable vector to transduce MSCs that are resistant to Ad5-mediated gene transfer. MSCs infected with AdF35 that activate an exogenous gene by the cytomegalovirus promoter can be a vehicle to deliver the gene product to targeted cells.