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In Vitro Dorsal Root Ganglia and Human Pancreatic Cancer Cell Line Interaction: A Co-culture Model

Author: LiuZhiSheng
Tutor: LiQiang；ZhangZuo
School: Tianjin Medical University
Course: Oncology
Keywords: Dorsal root ganglion Pancreatic Cancer Neurite Perineural infiltration Proliferation Apoptosis
CLC: R735.9
Type: Master's thesis
Year: 2008
Downloads: 62
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Abstract


Objective: pancreatic cancer (pancreatic cancer, PanCa) is a vicious one of the high degree of malignancy, early that happens lymphatics, blood vessels, liver metastases, the prognosis is poor. Perineural invasion (perineural invasion, PNI) is an important feature of pancreatic cancer, and retroperitoneal tumor recurrence is closely related with pancreatic cancer after an independent prognostic indicator of pancreatic cancer, however the PNI specific mechanism is unclear . Ayala used prostate cancer cell lines and rat dorsal root ganglia (dorsal root ganglion, DRGs) co-cultured prostate cancer cells / DRGs in vitro co-culture model, applied to the study of prostate cancer perineural infiltration, confirmed prostate cancer perineural The infiltration is related to prostate cancer cells with nerve cells or glial cells initiative interaction process. The purpose of this study is to establish a new in vitro pancreatic cancer perineural infiltration model and to observe the perineural infiltration process neurite growth condition, pancreatic cancer cell proliferation, apoptosis and migration. Methods: From 4 to 6-week-old Wistar rats neck, chest and waist take DRGs. Human pancreatic cancer cell lines (MIA PaCa-2) conventional cultivation, take the DRGs with human pancreatic cancer cell lines in Matrigel plastic culture research model. Inverted microscope observed at 24h, 48h, 72h, 96h, 120h and 144h pancreatic colony formation and of DRGs nerve cells neurite growth of pancreatic cancer cells migration and image analysis using Image-Pro Plus 5.0 image analysis software, neurite count, calculate the area occupied by neurites and pancreatic cancer cell colony. The immunocytochemistry assay of pancreatic cancer cells Ki-67 proliferation cell nuclear antigen expression, and calculate the pancreatic cancer cell proliferation rate (proliferation index, PI): the MTT assay absorbance value (A), reflect the proliferation of pancreatic cancer cells. Detection the apoptosis rate (apoptosis index.AI) of the pancreatic cancer cells by flow cytometry. Results: dorsal root ganglion neurons in the spinal cord at spinal ventral spinal nerve root, attached to the dorsal root, about the size of 1 to 2mm, pale yellow, round, smooth. Neurite outgrowth of nerve cells in the co-culture model was significantly better than the control group, after 72 hours, neurite occupies an area of ??approximately 2.57 times that of the control group (P <0.001); model MIA PaCa-2 pancreatic cancer cells co-cultured colony The growth rate is also faster than the control group, after 72 hours, the area occupied by the cell colony of approximately 1.33 times of the control group (P <0.001). Inverted microscope observed neurite obvious trend in the direction of the growth of pancreatic cancer cell colony, neurites and pancreatic cancer cell colony contact after a direction along the neurite growth of pancreatic cancer cells reverse migration, eventually reaching the dorsal root ganglia section around the PNI-like performance. Immunocytochemistry results show that the tumor cells, Ki-67 proliferating cell nuclear antigen expression were significantly higher in the supernatant culture group PI mean of the experimental group was 12.80 ± 1.84% (9.8% to 15.3%), while the control group PI average of 6.81 ± 0.73% (5.80% ~ 8.3%), the difference was statistically significant (P <0.001). MTT results showed that the absorbance of the experimental group was far higher than that in control group, the MTT and immune cell chemistry experiment results show that the pancreatic cancer cells in the pancreatic cancer cell proliferation in the experimental group than ordinary culture medium group active. Flow cytometry results showed that the supernatant culture group pancreatic cancer cell apoptosis was significantly lower than the control group and experimental group pancreatic cancer cells AI value of 2.46 ± 0.37% after 24 h incubation, while the control group AI value of 4.89 ± 0.63%, the difference was statistically significant (P <0.001). Conclusion: The human pancreatic cancer cell lines (MIA PaCa-2) / DRGs co-culture model was successfully constructed, to help pancreatic addicted to the study of neural invasion. Nerve cell - the pancreatic cell interactions play an important role in pancreatic cancer perineural infiltration process. Nerve cells and pancreatic cancer cells can promote each other's growth, and pancreatic cancer cells along the neurite growth the direction reverse support or oppose the trend of migration root ganglion direction. Many cytokines involved in this process, the molecular mechanism needs further study.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Pancreatic tumors
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