摘要
目的探讨胰岛素样生长因子结合蛋白7（IGFBP7）基因表达下调对急性髓系白血病细胞系U937细胞增殖及侵袭能力的影响。方法利用肝癌细胞株SMMC7721细胞，筛选最有效的IGFBP7干扰序列，瞬时转染处于对数生长期U937细胞，用Westernblot法检测转染后U937细胞IGFBP7蛋白表达。观察转染IGFBP7基因干扰序列后U937细胞的增殖、黏附、穿膜和浸润能力。结果IGFBP7基因干扰序列转染24h后U937细胞增殖能力明显下降，明显低于对照1组（未转染siRNA组）和对照2组（阴性干扰序列组）（A值分别为0．580±0．159、1．049±0．274、0．946±0．195）（P〈0．01）。IGFBP7基因下调后的U937细胞黏附于人脐静脉内皮细胞系ECV304细胞的数量明显低于对照1组和对照2组（A值分别为0．247±0．031对0．406±0．023和0．395±0．011）（P〈0．01）；穿越ECV304细胞包裹的细胞培养小室到底层的细胞明显减少[（0．387±0．021）×10^5对（1．017±0．031）×10^5、（0．908±0．027）×10^5]；实验组黏附于Matrigel基质膜的细胞数明显低于对照组[（0．197±0．098）×10^5对（0．493±0．067）×10^5和（0．469±0．083）×10^5]，差异有统计学意义（P值均〈0．01）。结论IGFBP7基因通过影响U937细胞增殖以及与基质内皮细胞的黏附、穿膜、侵袭能力而参与急性髓系白血病的发生、发展过程。
Objective To explore the effect of down-regulation of insulin-like growth factor binding protein 7 （ IGFBP7 ） on the proliferation and invasiveness of leukemia cell line U937 cells. Methods Three pairs of double-strand siRNA targeting IGFBP7 gene were transfected into SMMC7721 cells to select the most efficient one for U937 cells, qRT-PCR and Western blot were used to detect the expression of IGFBP7 in U937 cells after transiently transfected with siRNA of IGFBPT. Cell proliferation, adhesion, trans-endothelial migration and invasion were performed in transfected cells and control groups. Results After transfected with siRNA of IGFBP7 in U937 cells, the ability of cell proliferation was significantly decreased at 24 h （0. 580 ±0. 159） compared to that of parental cells and scramble negative control（ 1. 049 ±0.274, 0.946 ±0. 195, respectively） （P 〈0. 01 ）. Adhesion of U937 cells transfected with IGFBP7 gene specific siRNA to ECV304 cells was significantly lower than that of the control groups （ 0. 247 ± 0.031 vs 0. 406 ± 0. 023 and 0. 395 ±0. 011 ） （P 〈 0.01 ）. Transendothelial membrane of U937 cells into the bottom of the 24-well plate for experi- mental group were less than those in the control groups [ （0. 387 ±0. 021 ） ×10^5 vs （ 1. 017 ±0.031 ） ×10^5 and （0. 908 ±0. 027） ×10^5]. Cells adherent to the matrigel for experimental group were less than those in the control groups [ （0- 197 ± 0. 098） ×10^5 vs （0. 493 ±0. 067） ×10^5 and （0. 469 ＋ 0. 083 ） ×10^5 1. The difference was significant （ P 〈 0. 01 ）. Conclusion IGFBP7 gene plays a contributing role in leukemogenesis involving in leukemic cells＇ proliferation and interaction with endothelial cells through adhesion, invasion and migration.
出处
《中华血液学杂志》
CAS
CSCD
北大核心
2012年第4期307-310,共4页
Chinese Journal of Hematology
基金
江苏省自然科学基金（BK2009127）
江苏省卫生厅项目（H200921）
江苏省333高层次人才培养工程依托项目（H200921）
江苏省135开放课题项目（KF-20945）
江苏省高校自然基金（06KJB320101）
关键词
基因
IGFBP7
细胞系
U937
RNA干扰
Gene, insulin-like growth factor binding protein 7 （IGFBP7）
Cells line, U937
RNA interference
