[Effects of inhibiting SDF-1 expression by RNA interference on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells].
OBJECTIVE
To observe the effects of inhibiting stromal cell derived factor-1 (SDF-1) expression by RNA interference (RNAi) on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells.


METHODS
SDF-1 specific short hairpin RNA (shRNA) expressing plasmid was transferred into cultured human acute leukemic bone marrow stromal cells, positive clones were isolated by screening G418 resistance (Group A) , SDF-1 protein level in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The adhesion rates to bone marrow stromal cells layer and the drug sensitivity to doxorubicin of co-cultured Jurkat cells were detected by cell counting and MTT assay, respectively. The un-transfected bone marrow stromal cells of acute leukemia patient (Group B) or normal subject (Group C) were taken as control.


RESULTS
The level of secreted SDF-1 protein (pg/10(5) cells/week) in the supernatants of Group A, B and C were 1920 +/- 205, 12,370 +/- 1355 and 6620 +/- 770, respectively. Of co-cultured Jurkat cells in Group A, B and C, the adhesion rates after 24 h co-culturing were (28.8 +/- 2.6)%, (57.4 +/- 3.8)% and (45.2 +/- 4.0)%, respectively, and the IC50 values of doxorubicin were 585, 6162 and 1758 nmol/L, respectively.


CONCLUSIONS
Down-regulating SDF-1 expression of bone marrow stromal cells by RNAi reduces adhesion rates and enhances drug sensitivity to doxorubicin of their co-cultured Jurkat cells.
Yang
Wen-Bo
WB
Department of Hematology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
Kong
Pei-Yan
PY
Chang
Cheng
C
Wei
Li
L
Zeng
Dong-Feng
DF
Peng
Xian-Gui
XG
Shi
Zhan-Zhong
ZZ
Liu
Hong
H
Liu
Lin
L
Chen
Xing-Hua
XH
Wang
Qing-Yu
QY
D001854:Bone Marrow Cells / Q000378:metabolism*; D002448:Cell Adhesion; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000235:genetics* / Q000378:metabolism; D018920:Coculture Techniques; D019008:Drug Resistance, Neoplasm; D015870:Gene Expression; D006801:Humans; D019169:Jurkat Cells; D034622:RNA Interference; D017154:Stromal Cells / Q000378:metabolism*
D054377:Chemokine CXCL12