[Construction and tumorigenic study on a novel fusion gene AML1-MTG16].
OBJECTIVE
To test whether splicing overlapping extension(SOE) method can be a tool for obtaining rare fusion gene's transcripts and to study the tumorigenic capacity of a novel fusion gene AML1-MTG16.


METHODS
SOE method was used to obtain AML1-   MTG16 fusion gene's transcripts. MTG16, AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 segment were inserted into pEGFP-   C1,pDsRed-N1 vector respectively,then transfected NIH3T3 cell line by lipofection. Forty-eight hours later, the transfected cells were examined by laser-scanning confocal microscopy. Stable transfected cells were obtained by G418 500ug/ul selection for one month. Growth curve, soft agar colonies formation  tumorigenesis in nude mice were done to compare the difference between stable transfected cells.


RESULTS
Recombined AML1-MTG16 by SOE contained its CDS. NIH3T3 expressing AML1-MTG16 had a faster proliferation in medium, colony growth in soft agar. AML1-MTG16 expression cells also induced tumors formation following injection into nude mouse. MTG16,AML1-MTG16 and AML1-MTG16 without III,VI conserved domains of MTG16 were colocalized in the nucleus of cotransfected NIH3T3 cells under the examination of laser-scanning confocal microscope.


CONCLUSIONS
SOE is an effective method to get rare fusion gene's transcripts. AML1-MTG16 plays an important role in leukemogenesis. MTG16 may also have a carcinogenic property within the AML1-MTG16 fusion gene. Carcinogenic property of AML1-MTG16 is restricted to its localization in the nuclear matrix. N terminal of MTG16 may play an important part in the carcinogenic activity of AML1-MTG16.
Wang
Yang
Y
Shanghai Institute of Hematology, Ruijin Hospital of Shanghai Second Medical University, Shanghai,200025 P.R. China. zhugangw@shsmu.edu.cn
Lu
Shunyuan
S
Kong
Hui
H
Wang
Long
L
Yuan
Wentao
W
Sun
Yueping
Y
Jing
Yue'e
Y
Lu
Zhenyu
Z
Wang
Zhenyi
Z
Wang
Zhugang
Z
D016475:3T3 Cells / Q000637:transplantation; D000818:Animals; D002455:Cell Division / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D017690:Cell Transplantation; D050676:Core Binding Factor Alpha 2 Subunit; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D018613:Microscopy, Confocal; D009374:Neoplasms, Experimental / Q000235:genetics / Q000473:pathology; D015514:Oncogene Proteins, Fusion / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics*; D014162:Transfection
C112643:AML1-MTG16 protein, human; D050676:Core Binding Factor Alpha 2 Subunit; D008164:Luminescent Proteins; D015514:Oncogene Proteins, Fusion; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins