Influence of recombinant retroviral vector expressing antisense TGF alpha on malignant phenotype of human pancreatic carcinoma cell line.
OBJECTIVE
To observe the effects of antisense TGF alpha on the growth of human pancreatic carcinoma cell line cells.


METHODS
A recombinant retroviral vector expressing antisense TGF alpha was constructed, and transfected the ecotropic packaging cell line psi-2 with lipofectin. After the amphotropic packaging cell line PA317 was transfected with the virus supernatant of psi-2, the replication-defective, amphotropic retroviral supernatant was used to infect human pancreatic carcinoma cell line PC-7. Following puromycin selection, puromycin-resistant colonies were pooled and expanded to a cell line PC-7/AS-TGF alpha.


RESULTS
The retroviral integration in the genomes of psi-2, PA317 and transformant PC-7 cells was confirmed by Southern blot hybridization. Northern blot hybridization showed a down regulation of endogenous TGF alpha in PC-7/AS-TGF alpha cell line. The high levels of growth inhibition and reduction of 3H-TdR incorporation in PC-7/AS-TGF alpha were evident. Also, the soft agar colony-formation and tumorigenicity in nude mice were significantly suppressed by antisense TGF alpha.


CONCLUSIONS
The antisense TGF alpha expressing vector can block the target gene expression, suppress the cell growth and partially reverse the malignant phenotype of pancreatic carcinoma cells.
Xu
Y
Y
Department of Pathology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China.
Liu
T
T
Gao
J
J
D016475:3T3 Cells / Q000473:pathology; D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology*; D000818:Animals; D002455:Cell Division; D015536:Down-Regulation; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology*; D016372:RNA, Antisense; D014162:Transfection; D016211:Transforming Growth Factor alpha / Q000096:biosynthesis / Q000235:genetics*; D014407:Tumor Cells, Cultured
D016372:RNA, Antisense; D016211:Transforming Growth Factor alpha