lipofectamine and Pancreatic-Neoplasms

To explore the expression and biological function of protocadherin 10 (PCDH10) in pancreatic cancer cells.. Reverse transcription PCR (RT-PCR) was used to detect the expression of PCDH10 in CAPAN-1, PANC-1, ASPC-1, BXPC-3 pancreatic cancer cells and the HPDE6-C7 normal human pancreatic ductal epithelial cells. Recombinant plasmid pcDNA3.1-PCDH10 and empty vector pcDNA3.1 were transfected into BXPC-3 pancreatic cancer cells via Lipofectamine(TM)2000. After transfection, the mRNA expression of PCDH10 was examined by RT-PCR, and the protein level was detected by Western blotting. CCK-8 and colony formation assays were used to analyze the cell proliferation. Cell apoptosis was tested by flow cytometry combined with annexin V-FITC/PI staining. Transwell(TM) and wound healing assays were performed to measure the invasion and migration ability of the cells.. Compared with the normal pancreatic ductal epithelial cells, the expression of PCDH10 was obviously lower in the CAPAN-1, PANC-1, BXPC-3 cells. RT-PCR and Western blotting revealed that PCDH10 expression significantly increased in BXPC-3 cells transfected with plasmid pcDNA3.1-PCDH10 compared with the ones with empty vector pcDNA3.1. CCK-8 and colony formation assays showed that the PCDH10-transfected cells grew more slowly than the empty vector-transfected cells. Annexin V-FITC/PI staining combined with flow cytometry proved that the apoptosis in the PCDH10-transfected cells remarkably increased compared with that in the control group. A reduction of the invasion and migration ability was found obviously in the PCDH10-transfected cells by Transwell(TM) assay. The wound healing assay also showed that the PCDH10-transfected cells spread the more slowly than the empty vector-transfected cells.. The expression of PCDH10 was down-regulated in the pancreatic cancer cells. PCDH10 over-expression could significantly induce cell apoptosis, and restrain proliferation, invasion and migration ability of BXPC-3 pancreatic cancer cells.

Topics: Apoptosis; Blotting, Western; Cadherins; Cell Line; Cell Line, Tumor; Cell Movement; Cell Proliferation; Down-Regulation; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Lipids; Neoplasm Invasiveness; Pancreatic Neoplasms; Protocadherins; Reverse Transcriptase Polymerase Chain Reaction; Transfection

To investigate the toxicity of cationic liposome Lipofectamine 2000 (Lipo) in human pancreatic cancer Capan-2 cells.. Capan-2 cells were cultured in the presence of Lipo at toxic concentrations, and the cell growth, apoptosis and cell cycle changes were evaluated by cell counting and flow cytometry.. The concentrations of both Lipo and siRNA affected the transfection efficiency. In a transfection volume of 2 ml, the presence of 5 microl Lipo resulted in slowed growth of Capan-2 cells, which was especially obvious after 3 days (P<0.001). Prolonged culture of the transfected cells caused significant increases in early apoptotic cells (P<0.05) and in the damaged or necrotic cells (P<0.001), and resulted in reduced viable cells (P<0.01); these changes became obvious after a 48-hour culture, which also increased the ratio of G(0)/G(1) phase cells (P<0.05) and decreased those of G(2)/M phase cells (P<0.01), S phase cells (P<0.01), and the late apoptotic cells (P<0.05).. Toxic concentrations of Lipo can affect the growth, apoptosis and cell cycles of Capan-2 cells in vitro, and this urges careful concentration selection when using Lipo for gene transfer into different cells.

Topics: Apoptosis; Cations; Cell Cycle; Cell Line, Tumor; Humans; Lipids; Liposomes; Pancreatic Neoplasms; RNA, Small Interfering; Transfection