CAJ | 학술논문

目的：构建血小板源性生长因子B（PDGF-B）慢病毒载体，建立PDGF-B稳定过表达的 SGC7901胃癌细胞株并观察PDGF-B对 SGC7901胃癌细胞侵袭能力的影响。方法采用四质粒系统构建 PDGF-B慢病毒载体，转染293T细胞包装获得高滴度病毒颗粒；以病毒颗粒转染SGC7901细胞，通过荧光显微镜观察转染效果，并用免疫印迹法检测PDGF-B的蛋白表达；用Tr-answell试验观察 SGC7901细胞侵袭能力的变化。结果通过PCR及酶切鉴定成功构建 PDGF-B载体；包装并得到高滴度的病毒颗粒；成功转染 SGC7901细胞后，免疫印迹法检测到 PDGF-B蛋白表达明显升高；Transwell试验证明 PDGF-B过表达增强SGC7901细胞侵袭能力。结论成功构建并包装获得PDGF-B慢病毒颗粒，建立 PDGF-B稳定过表达 SGC7901胃癌细胞株，证明PDGF-B过表达可增强胃癌细胞侵袭转移能力。
목적：구건혈소판원성생장인자B（PDGF-B）만병독재체，건립PDGF-B은정과표체적 SGC7901위암세포주병관찰PDGF-B대 SGC7901위암세포침습능력적영향。방법채용사질립계통구건 PDGF-B만병독재체，전염293T세포포장획득고적도병독과립；이병독과립전염SGC7901세포，통과형광현미경관찰전염효과，병용면역인적법검측PDGF-B적단백표체；용Tr-answell시험관찰 SGC7901세포침습능력적변화。결과통과PCR급매절감정성공구건 PDGF-B재체；포장병득도고적도적병독과립；성공전염 SGC7901세포후，면역인적법검측도 PDGF-B단백표체명현승고；Transwell시험증명 PDGF-B과표체증강SGC7901세포침습능력。결론성공구건병포장획득PDGF-B만병독과립，건립 PDGF-B은정과표체 SGC7901위암세포주，증명PDGF-B과표체가증강위암세포침습전이능력。
Objective To construct a lentiviral vector that stably express platelet-derived growth factor B (PDGF-B)and to in-vestigate effect of PDGF-B overexpression on invasion of SGC7901 gastric carcinoma cell.Methods The lentiviral vector pLeno-DCE-PDGF-B was constructed and transfected into 293T cells.The supernatant containing the lentivirus particles was harvested to determine the virus titer and high titer lentivirus particles was gathered.Then PDGF-B lentiviral vector was transfected into SGC7901 cells for construction of stable PDGF-B overexpression SGC7901 cells.Western-blot and immunofluorescence were used for evaluation of the construction of stable PDGF-B overexpression SGC7901 cells.Transwell test was used for detection the inva-sion of SGC7901 cells.Results The lentiviral vector was correctly constructed and verified by sequencing.High titer PDGF-B lenti-viral particles were acquired successfully.After transfection,SGC7901 cells were green color by detection of fluorescence micro-scope;and the expression of PDGF-B protein in transfected SGC7901 cells were much higher than that in normal SGC7901 cells.Al-so,PDGF-B overexpression increased the invasion ability of SGC7901 cells.Conclusion The PDGF-B lentiviral particles are ob-tained after successful construction and package.The stable PDGF-B overexpression SGC7901 gastric carcinoma cell line is estab-lished,which proves that the PDGF-B overexpression might increase the invasion ability of gastric carcinoma cells.