CAJ | 학술논문

目的：研究Numb基因对人肾癌细胞侵袭和迁移能力的影响及其相关机制。方法选取人肾癌细胞786-O为研究对象，采用Numb-ORF表达质粒转染细胞为实验组，设置阴性对照组及空白对照组。采用荧光定量聚合酶链反应（PCR）和蛋白质印迹技术检测各组中Numb和基质金属蛋白酶9（MMP-9）的表达情况，采用细胞划痕实验和Transwell小室法分别检测细胞的迁移和侵袭能力。结果实验组中Numb的相对表达水平为36.13±8.33，阴性对照组为2.51±0.27，空白对照组为2.87±0.21，差异有统计学意义（P＝0.00）；实验组MMP-9相对表达水平为2.36±0.29，阴性对照组为8.73±0.91，空白对照组为8.99±0.78，差异有统计学意义（P＝0.00）。划痕实验中，12 h划痕修复比为：实验组0.53±0.06，阴性对照组0.73±0.09，空白对照组0.75±0.08，差异有统计学意义（P＝0.02）。侵袭实验中，实验组的穿膜细胞数为（31.40±5.96）个，少于阴性对照组的（126.93±13.61）个和空白对照组的（131.87±15.42）个，差异有统计学意义（P＝0.00）。结论上调Numb基因可以降低人肾癌细胞786-O的侵袭和迁移能力，该作用可能是通过Numb负性调节MMP-9实现的。
목적：연구Numb기인대인신암세포침습화천이능력적영향급기상관궤제。방법선취인신암세포786-O위연구대상，채용Numb-ORF표체질립전염세포위실험조，설치음성대조조급공백대조조。채용형광정량취합매련반응（PCR）화단백질인적기술검측각조중Numb화기질금속단백매9（MMP-9）적표체정황，채용세포화흔실험화Transwell소실법분별검측세포적천이화침습능력。결과실험조중Numb적상대표체수평위36.13±8.33，음성대조조위2.51±0.27，공백대조조위2.87±0.21，차이유통계학의의（P＝0.00）；실험조MMP-9상대표체수평위2.36±0.29，음성대조조위8.73±0.91，공백대조조위8.99±0.78，차이유통계학의의（P＝0.00）。화흔실험중，12 h화흔수복비위：실험조0.53±0.06，음성대조조0.73±0.09，공백대조조0.75±0.08，차이유통계학의의（P＝0.02）。침습실험중，실험조적천막세포수위（31.40±5.96）개，소우음성대조조적（126.93±13.61）개화공백대조조적（131.87±15.42）개，차이유통계학의의（P＝0.00）。결론상조Numb기인가이강저인신암세포786-O적침습화천이능력，해작용가능시통과Numb부성조절MMP-9실현적。
Objective To study the effect of Numb gene overexpression on invasion and migration in human renal cell carcinoma cells and its related mechanism. Methods The Numb-ORF plasmid was transfected into renal cell carcinoma cells 786-O, set the negative control and blank control. The expression of Numb and matrix metalloproteinase-9 (MMP-9) was detected by real-time PCR and western blot. Then abilities of cells invasion and migration were measured respectively by transwell assay and scratch-wound-healing assay. Results Compared to negative control (2.51±0.27) and blank control (2.87±0.21), Numb expression in Numb-ORF group (36.13 ±8.33) was significantly higher (P= 0.00). Meanwhile, MMP-9 level in Numb-ORF group was decreased: Numb-ORF group was 2.36±0.29, negative control was 8.73±0.91, blank control was 8.99±0.78 (P=0.00). In scratch-wound-healing assay,the repair ratio of 12 h was as follow: Numb-ORF group was 0.53 ±0.06, negative control was 0.73 ±0.09, blank control was 0.75 ±0.08 (P= 0.02). In transwell assay, the number of invasion cells in Numb-ORF group was 31.40±5.96, less than negative control (126.93±13.61) and blank control (131.87 ±15.42) (P= 0.00). Conclusion Overexpression of Numb significantly decreases abilities of invasion and migration in 786-O cells, and the suppressive effect may be due to the down-regulation of MMP-9 expression in human renal cell carcinoma.