[The effect of recombinant adenovirus encoding antisense MAT1 on cell cycle of human pancreatic cancer cells BxPC-3].
OBJECTIVE
To investigate the regulatory effect of the human MAT1 gene on the cell cycle G(1)/S transition of human pancreatic cancer BxPC-3 cells.


METHODS
To construct the replication deficient recombinant adenovirus of antisense MAT1 gene using homologous recombination by AdEasy system. Cell growth assay was carried out by counting alive cells after trypan blue exclusion. The protein expressions of MAT1, cyclin D1, cyclin E, and Rb were detected by western blotting. The cell cycle status was determined by flow cytometry.


RESULTS
The recombinant adenovirus encoding antisense MAT1 fragment Ad-MAT1AS was obtained with the titer of expression was 5 x 10(10) pfu/ml. The expression of MAT1 of BxPC-3 was significantly reduced after Ad-MAT1AS infection. In this case BxPC-3 cell cycle was arrested in G(1) phase. The estimated proportion of G(0)/G(1) phase cells in the control for blank and vector cultures ranged from 40% to 44%. In contrast, 79% of the Ad-MAT1AS cells were in G(0)/G(1) phase. Cyclin E and pRb gene expression changes were observed in the infected cells.


CONCLUSIONS
The results suggest that MAT1 gene may play an important role in the regulation of cell cycle G(1)-->S transition of BxPC-3 cells.
Zhang
Shi-neng
SN
Department of Gastroenterology, the Second Affiliated Hospital of Sun Yat-sen University, Guangzhou 510120, China.
Xu
Feng-qin
FQ
Huang
Zhi-qing
ZQ
Zeng
Zhi-yong
ZY
Yuan
Shi-zhen
SZ
D000256:Adenoviridae / Q000235:genetics; D002453:Cell Cycle; D018844:Cyclin-Dependent Kinases / Q000235:genetics* / Q000378:metabolism*; D016213:Cyclins / Q000378:metabolism*; D006801:Humans; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured; D000095043:Cyclin-Dependent Kinase-Activating Kinase
D016213:Cyclins; D020319:Oligodeoxyribonucleotides, Antisense; D011994:Recombinant Proteins; D018844:Cyclin-Dependent Kinases; D000095043:Cyclin-Dependent Kinase-Activating Kinase