Antisense oligonucleotide targeting endostatin enhances hematopoiesis reconstitution in BMT mice.
- Author:
Ning WU
1
;
Jie-lin QI
;
De-rong HU
;
Xi-qin ZHANG
;
Bing BU
;
Zhi-fang LIU
;
Han-ying SUN
;
Wen-li LIU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Bone Marrow Cells; drug effects; metabolism; Bone Marrow Transplantation; Dose-Response Relationship, Drug; Endostatins; biosynthesis; genetics; Female; Hematopoiesis; Intercellular Adhesion Molecule-1; biosynthesis; genetics; Male; Mice; Mice, Inbred BALB C; Oligonucleotides, Antisense; pharmacology; RNA, Messenger; biosynthesis; Reverse Transcriptase Polymerase Chain Reaction; Transfection
- From: Chinese Journal of Hematology 2006;27(8):534-537
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effect of antisense oligonucleotide targeting endostatin (endostatin-ASON) transfecting bone marrow stromal cells ( BMSC) on hematopoiesis reconstitution in BMT mice.
METHODSInhibition of endostatin / VCAM-1 protein and mRNA expression was investigated by transfection of antisense oligonucleotide targeting endostatin with confocal microscopy, Western blot and RT-PCR. Bone marrow stromal cells were cultured and divided into 4 groups: group (1) without any treatment; group (2) BMT only; group (3) BMT + endostatin-ASON transfection; group (4) BMT + endostatin scrambled sequence transfection.
RESULTS(1) Endostatin-ASON was successfully introduced into BMSC in vitro, and the transfecting rate was 86% ;(2) After Endostatin-ASON transfected into BMSC, the expression of Endostatin mRNA and its protein on the BMSC was signficantly inhibited at different time point after BMT [the grey value of Endostatin was (0.09 +/- 0.03) - (1.44 +/- 1.19) and (0.02 + 0.02) - (0.14 +/- 0.05), respectively] (P < 0.01 and P < 0.05); (3) Transfecting with Endostatin-ASON effectively promoted the expression of VCAM-1 mRNA and its protein on the BMSC [the gray value of VCAM-1 was (1.60 +/- 0. 92) - (8.05 +/- 0.87) and (0.07 +/- 0.02) - (0.67 +/- 0.09) , respectively] (P <0.01 and P <0.05) ; (4) There was no effects of transfecting Endostatin scrambled sequence on the expression of Endostatin and VCAM-1 on the BMSC (P > 0.05).
CONCLUSIONEndostatin-ASON could inhibit Endostatin expression and enhance VCAM-1 expression in BMSC after syngeneic-BMT in mice, which might be one of the mechanisms underlying the endostatin-ASON accelerating hematopoiesis reconstitution after allogeneic-BMT.
