摘要
Abstract Objective To observe the effects of antisense TGFα on the growth of human pancreatic carcinoma cell line cells. Methods A recombinant retroviral vector expressing antisense TGFα was constructed, and transfected the ecotropic packaging cell line ψ 2 with lipofectin. After the amphotropic packaging cell line PA317 was transfected with the virus supernatant of ψ 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 TGFα. Results The retroviral integration in the genomes of ψ 2, PA317 and transformant PC 7 cells was confirmed by Southern blot hybridization. Northern blot hybridization showed a down regulation of endogenous TGFα in PC 7/AS TGFα cell line. The high levels of growth inhibition and reduction of 3 H TdR incorporation in PC 7/AS TGFα were evident. Also, the soft agar colony formation and tumorigenicity in nude mice were significantly suppressed by antisense TGFα. Conclusion The antisense TGFα expressing vector can block the target gene expression, suppress the cell growth and partially reverse the malignant phenotype of pancreatic carcinoma cells.
Abstract Objective To observe the effects of antisense TGFα on the growth of human pancreatic carcinoma cell line cells. Methods A recombinant retroviral vector expressing antisense TGFα was constructed, and transfected the ecotropic packaging cell line ψ 2 with lipofectin. After the amphotropic packaging cell line PA317 was transfected with the virus supernatant of ψ 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 TGFα. Results The retroviral integration in the genomes of ψ 2, PA317 and transformant PC 7 cells was confirmed by Southern blot hybridization. Northern blot hybridization showed a down regulation of endogenous TGFα in PC 7/AS TGFα cell line. The high levels of growth inhibition and reduction of 3 H TdR incorporation in PC 7/AS TGFα were evident. Also, the soft agar colony formation and tumorigenicity in nude mice were significantly suppressed by antisense TGFα. Conclusion The antisense TGFα expressing vector can block the target gene expression, suppress the cell growth and partially reverse the malignant phenotype of pancreatic carcinoma cells.
