[Cloning and functional analysis of the tumor metastasis related gene tropomyosin2].
OBJECTIVE
To construct an eukaryotic expression vector of tumor associated gene tropomyosin 2 (TM2) tagged with green fluorescence protein and study its biological role in tumorigenesis. METHODS; Whole length of TM2 cDNA sequence was amplified from human gastric cancer cell line AGS by reverse transcript PCR. The recombinant TM2 was inserted into eukaryotic expression vector pIRES2 tagged with enhanced green fluorescence protein. pIRES2-EGFP was transfected into human hepatoma cell line QGY-7703 cells by liposome technique and detected by fluorescence microscopy and flow cytometry. TM2 protein level was determined by Western blot. The in vitro invasion and motility were further studied. Its tumorigenesis was assessed on nude mice.


RESULTS
The recombinant TM2 was shown to be expressed in QGY-7703 cells by fluorescence microscopy and flow cytometry. The protein level of TM2 was increased. The invasion ability of TM2- transfected cells was increased(45.6 +/- 9.9) than that in controls(21.6 +/- 3.3, P < 0.05), as well as mobility(41.4 +/- 11.8 vs. 16.7 +/- 3.7). The tumor volume was (241.5 +/- 95.1) mm3, significantly higher than that in blank-vector controls (100.6 +/- 85.4) mm3 and negative-controls (123.7 +/- 92.3) mm3.


CONCLUSIONS
TM2 plays a role in growth and metastasis of hepatic tumor.
Zhang
Yi
Y
Department of Surgery, Ruijin Hospital, Shanghai Jiao Tong University, Shanghai Institute of Digestive Surgery, Shanghai 200025, China.
Liu
Wen-Tao
WT
Li
Qiang
Q
Chen
Xue-Hua
XH
Liu
Bing-Ya
BY
Zhu
Zheng-Gang
ZG
D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D018076:DNA, Complementary / Q000235:genetics; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009368:Neoplasm Transplantation; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013274:Stomach Neoplasms / Q000235:genetics / Q000473:pathology; D014162:Transfection; D014335:Tropomyosin / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D047368:Tumor Burden
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D014335:Tropomyosin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins