[Effects of in vitro PTEN transfection on proliferation of human pancreatic cancer ASPC-1 cells].
Hongyu Li, Jie‐Ping Yu, Xiaozhong Guo, Jianjun Li, Jiajun Zhao, Di Wang, Ran Huo, Xia Yu-ting
Abstract
Hongyu Li, Jie‐Ping Yu, Xiaozhong Guo, Jianjun Li, Jiajun Zhao, Di Wang, Ran Huo, Xia Yu-ting
Abstract
OBJECTIVE: To investigate the effect of phosphatase and ensinhomology deleted on chromosome ten (PTEN) on the human pancreatic cancer cell line without or low PTEN expression. METHODS: We selected the lowest PTEN gene expressive pancreatic cancer cell line among the four pancreatic cancer cell lines (Miapaca I, Miapaca II, JF305 and ASPC-1) through RT-PCR assay method, and transfected plasmid (Peak8) inserting PTEN or not in vitro into it by lipofectin. The effect of PTEN transfection on the lowest PTEN gene expressive pancreatic cancer cell line was carried out by flow cytometry, immunohistochemical staining, cloning survival assay, as well as tumorigenicity in nude mice. RESULTS: The lowest PTEN gene expression was found in ASPC-1 cells. PTEN mRNA expression and cloning plating efficiency in the ASPC-1, A-pE and A-pE-P cells were 20.3%, 15.0%, 56.8% and 33.3%, 31.7%, 24.0% respectively. We found that ASPC-1 cells transfected with PTEN exhibited significantly more protein, less vascular endothelial growth factor protein, and non-changed epidermal growth factor receptor protein comparing with vector-transfected cells. The tumor volumes were 202.7 mm(3) and 142.4 mm(3), pre-and post-transfection with significance (P < 0.01) within 5 weeks. CONCLUSIONS: Our findings suggested that PTEN mRNA exhibited the lowest expression in ASPC-1 cell line, exogenous PTEN dramatically inhibits the growth of ASPC-1 cells transfected with PTEN gene in nude mice.
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OBJECTIVE: To investigate the effect of phosphatase and ensinhomology deleted on chromosome ten (PTEN) on the human pancreatic cancer cell line without or low PTEN expression. METHODS: We selected the lowest PTEN gene expressive pancreatic cancer cell line among the four pancreatic cancer cell lines (Miapaca I, Miapaca II, JF305 and ASPC-1) through RT-PCR assay method, and transfected plasmid (Peak8) inserting PTEN or not in vitro into it by lipofectin. The effect of PTEN transfection on the lowest PTEN gene expressive pancreatic cancer cell line was carried out by flow cytometry, immunohistochemical staining, cloning survival assay, as well as tumorigenicity in nude mice. RESULTS: The lowest PTEN gene expression was found in ASPC-1 cells. PTEN mRNA expression and cloning plating efficiency in the ASPC-1, A-pE and A-pE-P cells were 20.3%, 15.0%, 56.8% and 33.3%, 31.7%, 24.0% respectively. We found that ASPC-1 cells transfected with PTEN exhibited significantly more protein, less vascular endothelial growth factor protein, and non-changed epidermal growth factor receptor protein comparing with vector-transfected cells. The tumor volumes were 202.7 mm(3) and 142.4 mm(3), pre-and post-transfection with significance (P < 0.01) within 5 weeks. CONCLUSIONS: Our findings suggested that PTEN mRNA exhibited the lowest expression in ASPC-1 cell line, exogenous PTEN dramatically inhibits the growth of ASPC-1 cells transfected with PTEN gene in nude mice.
Key concepts: PTEN, Transfection, Cancer research, Pancreatic cancer, Cell culture, Molecular biology, Medicine, Cell growth