CAJ | 학술논문

目的：探讨小干扰RNA（ siRNA）技术沉默PI3Kp85α基因表达对人胃癌细胞株AGS增殖、迁移及侵袭能力的影响。方法设计3条靶向抑制PI3Kp85α基因的siRNA片段（ siRNA1、siRNA2、siRNA3），分别转染人胃癌AGS细胞，并设阴性对照组及空白组。 Western blotting检测siRNA序列对PI3Kp85α蛋白表达的影响。利用MTT实验、划痕迁移实验和Transwell侵袭实验检测下调 PI3Kp85α水平对 AGS 细胞增殖、迁移和侵袭能力的影响。结果转染 siRNA1、siRNA2和siRNA3的AGS细胞中PI3Kp85α蛋白水平均低于阴性对照组及空白组（ P＜0?05）。转染siRNA3的AGS细胞PI3Kp85α蛋白表达降低最明显，抑制率达80％，故选择siRNA3进行后续实验。转染24h后，PI3Kp85α siRNA转染组细胞增殖能力明显低于对照组细胞（P＜0?05）。24h体外划痕实验显示，PI3Kp85α siRNA转染组的划痕愈合能力明显低于对照组（P＜0?05）。Transwell实验显示，PI3Kp85α siRNA转染组细胞的细胞穿膜数明显低于对照组（ P＜0?05）。结论通过siRNA能明显下调PI3Kp85α蛋白在胃癌细胞AGS中的表达，并抑制肿瘤细胞的增殖、迁移和侵袭。
목적：탐토소간우RNA（ siRNA）기술침묵PI3Kp85α기인표체대인위암세포주AGS증식、천이급침습능력적영향。방법설계3조파향억제PI3Kp85α기인적siRNA편단（ siRNA1、siRNA2、siRNA3），분별전염인위암AGS세포，병설음성대조조급공백조。 Western blotting검측siRNA서렬대PI3Kp85α단백표체적영향。이용MTT실험、화흔천이실험화Transwell침습실험검측하조 PI3Kp85α수평대 AGS 세포증식、천이화침습능력적영향。결과전염 siRNA1、siRNA2화siRNA3적AGS세포중PI3Kp85α단백수평균저우음성대조조급공백조（ P＜0?05）。전염siRNA3적AGS세포PI3Kp85α단백표체강저최명현，억제솔체80％，고선택siRNA3진행후속실험。전염24h후，PI3Kp85α siRNA전염조세포증식능력명현저우대조조세포（P＜0?05）。24h체외화흔실험현시，PI3Kp85α siRNA전염조적화흔유합능력명현저우대조조（P＜0?05）。Transwell실험현시，PI3Kp85α siRNA전염조세포적세포천막수명현저우대조조（ P＜0?05）。결론통과siRNA능명현하조PI3Kp85α단백재위암세포AGS중적표체，병억제종류세포적증식、천이화침습。
Objective To investigate the influence of small interfering RNA( siRNA) targeting PI3Kp85α on proliferation, migration and invasion of gastric carcinoma cell line AGS. Methods Three siRNA with different sequences ( siRNA-1,siRNA-2,siR-NA-3) were desinged and transfected into AGS cells. The AGS cells were divided into siRNA groups, universal scrambled negative siRNA groups and blank groups. After transfection, the protein expression of PI3Kp85αwas detected by western blotting. MTT assay, wound healing and Transwell assays were performed to detect the effect of PI3Kp85α siRNA on AGS cell proliferation, migration and invasion. Results The expressions of PI3Kp85α protein significantly decreased in the siRNA-2 and siRNA-3 transfected cells. Western blotting analysis showed that cells transfected with siRNA-3 had the strongest inhibition of PI3Kp85αprotein, with the inhibi-tion rate being 80%, and siRNA-3 was chosen in the subsequence experiments for gene knockdown. MTT assay and wound healing as-say showed that the proliferative and healing ability of PI3Kp85α-knockdown cells was significantly lower than that of the negative con-trol cells since 24 hours after transfection, respectively(P<0?05). Transwell assay showed that the cells passing the polycarbonate membranes was significantly less in PI3Kp85α-knockdown cells than in the negative control cells (P<0?05). Conclusion siRNA down-regulating PI3Kp85α gene in human gastric carcinoma cell line AGS can inhibit cell proliferation,migration and invasion.