| Pancreatic adenocarcinoma is one of the most common malignant with poor prognosis and early metastasis. When diagnosed, 80% patients with pancreatic carcinoma have metastasized. And in the past 20 years the incidence of pancreatic cancer has increased one to fivefold and the overall 5-year survival rate of the patients is less than 1-5%. Although recently the operation is still an effective treatment for pancreatic carcinoma, mean survival time is 17 to 20 months in post-opration in pancreatic cancer. Therefore the striving with development, growthing machnism and signal conduction is always the focus of researchs. The study of related genediagnosis and therapy may provide further opportunities for innovative therapeutic intervention of pancreatic cancer.DPC4 inactivateing in nearly one-half of the pancreatic carcinomas is found a newly suppression gene. It has been reported to be involved in and played a key role in the TGF- β signal pathway. Its inactivateing interrupted the TGF- 3 signal pathway and resulted in the cells proliferation abnormally. Therefore studying the function of DPC4 gene will reveal the genesis and development mechanism of pancreatic carcinoma.In order to discusse the mechanism of inhibitory effect of DPC4 on pancreatic carcinoma growth and provide the experimental foundation for gene therapy of pancreatic carcinoma. we have investigated the role of DPC4 in inhibiting the growth and proliferation in pancreatic cancer line, as well as the relationship of DPC4, downstream gene P21 and Bcl-2, Bax gene related to apoptosis.The full length coding sequence of DPC4 gene was transfected into human pancreatic adenocarcinoma cell line JF305 using lipofectamine transfecting technique. Stable transfectants were obtained through G418(400ug/ml) selective culture for five weeks.The cells no- transfected and pBK-CMV transfected cell were used as controls. The expression of DPC4 and p21 were studied by flow cytometry and western blot. The expression of Bcl-2 and Bax were detected by flow cytometry. The cell growth rate was estimated by MTT method.Only JF305 cells transfected with pBK-CMV-DPC4 plasmid and pBK-CMV plasmid still survived after selected by culturing with G418. The... |
