[Effect of recombinant adenovirus carrying NLS-RARalpha gene on the proliferation of HL-60 cell and the differentiation of HL-60 cells induced by ATRA and relevant mechanism].
OBJECTIVE
To explore the effect and mechanism of recombined adenovirus carrying NLS-RARalpha gene on proliferation of HL-60 cells and the differentiation of HL-60 cells induced by ATRA.


METHODS
HL-60 cells was infected with Ad-NLS-RARalpha and control virus Ad-KZ. The efficiency of infection was detected by FCM. The mRNA and protein levels of NLS-RARalpha were assessed by Real-time PCR (RT-PCR) and Western blot, respectively. MTT assay were applied to determine proliferation of HL-60 cells. Cell surface differentiation antigen CD11b of infected HL-60 cell induced by ATRA was examined by FCM. The mRNA and protein levels of C-MYC of infected HL-60 cell induced by ATRA were determined by Real-time PCR (RT-PCR) and Western blot assay.


RESULTS
The efficiency of infection of Ad-NLS-RARalpha and Ad-KZ on HL-60 cell was 70%-80%. The mRNA and protein levels of NLS-RARalpha gene of HL-60 cells which infected with Ad-NLS-RARalpha were both obviously higher than that of the cells which infected with Ad-KZ and non-infected (P < 0.05). The proliferation ability of HL-60 cell infected with Ad-NLS-RARalpha was significantly increased (P < 0.05). The level of CD11b of HL-60 cell infected with Ad-NLS-RARalpha and induced by ATRA was clearly decreased than control groups (P < 0.05). The mRNA and protein levels of C-MYC gene of HL-60 cells infected with Ad-NLS-RARalpha and induced by ATRA were both obviously higher than that of the cells which infected with Ad-KZ and non-infected (P < 0.05).


CONCLUSIONS
The recombined adenovirus Ad-NLS-RARalpha can increase the proliferation ability of HL-60 cell, and inhibit the differentiation of HL-60 cell through reduce the expression level of C-MYC gene.
Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Chongqing 402160, China.
Xiu-Xiu
XX
Hu
Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Chongqing 402160, China.
Bei-Zhong
BZ
Liu
Key Laboratory of Laboratory of Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Liang
L
Zhong
Key Laboratory of Laboratory of Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Yuan-Mei
YM
Gao
Key Laboratory of Laboratory of Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Xi
X
Zhang
Key Laboratory of Laboratory of Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Xiu-Juan
XJ
Wu
Key Laboratory of Laboratory of Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Hui
H
Wang
Key Laboratory of Laboratory of Medical Diagnostics of Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
Xin-Yu
XY
Zhu
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000187:drug effects*; D018922:HL-60 Cells; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D018168:Receptors, Retinoic Acid / Q000235:genetics*; D000072482:Retinoic Acid Receptor alpha; D014212:Tretinoin / Q000494:pharmacology*; D028901:alpha Karyopherins / Q000235:genetics*
C000607071:RARA protein, human; D018168:Receptors, Retinoic Acid; D000072482:Retinoic Acid Receptor alpha; D028901:alpha Karyopherins; D014212:Tretinoin