CAJ | 학술논문

目的：构建稳定表达Rspo1的基因转染间充质干细胞株。方法将人Rspo1全长cDNA重组入逆转录病毒载体pEGZ-Term，通过与辅助病毒载体共转染293T细胞，包装为具有感染力的完整重组病毒载体，收集培养上清感染间充质干细胞C3H10 T1/2细胞株，筛选获得G418抗性的基因转染细胞，通过RT-PCR与流式细胞术检测感染后C3H10 T1/2细胞Rspo1分子的表达，并采用细胞计数法观察其上清对SW480结肠癌细胞增殖能力的影响。结果成功构建pEGZ-Term/Rspo1逆转录病毒表达载体，获得了稳定表达人Rspo1的基因转染细胞株C3H10/Rspo1，该细胞株上清能促进SW480结肠癌细胞增殖。结论建立了稳定表达Rspo1的基因转染间充质干细胞株，为进一步利用Rspo1和间充质干细胞靶向作用于肠道干细胞救治肠上皮损伤的研究奠定了基础。
목적：구건은정표체Rspo1적기인전염간충질간세포주。방법장인Rspo1전장cDNA중조입역전록병독재체pEGZ-Term，통과여보조병독재체공전염293T세포，포장위구유감염력적완정중조병독재체，수집배양상청감염간충질간세포C3H10 T1/2세포주，사선획득G418항성적기인전염세포，통과RT-PCR여류식세포술검측감염후C3H10 T1/2세포Rspo1분자적표체，병채용세포계수법관찰기상청대SW480결장암세포증식능력적영향。결과성공구건pEGZ-Term/Rspo1역전록병독표체재체，획득료은정표체인Rspo1적기인전염세포주C3H10/Rspo1，해세포주상청능촉진SW480결장암세포증식。결론건립료은정표체Rspo1적기인전염간충질간세포주，위진일보이용Rspo1화간충질간세포파향작용우장도간세포구치장상피손상적연구전정료기출。
Objective To construct a gene transfected mesenchymal stem cell line that stably expressed Rspo1. Methods The full length human Rspo1 cDNA was subcloned into retroviral expressing vector pEGZ-Term. The recombinant plasmid together with its helper virus vector was cotransfected into the package cell 293T. The C3H10 T1/2 cells were infected with the supernatant of the transfected 293T cells, and then were selected with G418. The G418 resistant cells were harvested for screening their C3H10/Rspo1 expression by RT-PCR and flow cytometry. The biological effect of supernatant was analyzed by cell counting. Results The pEGZ-Term/Rspo1 retrovirus expressing vector was constructed successfully and a cell line, named C3H10/Rspo1, which sta-bly expressed Rspo1 was obtained and its supernatant could promote colon cancer cell line SW480 proliferation. Conclusion In our study, a stable gene transfected mesenchymal stem cell line that expressed Rspo1 was estab-lished. It provides a valuable tool to explore the novel strategy to rescues the intestinal epithelium damage.