Application of fibroblast activation protein (FAP) alpha to preparation of pancreatic cancer prognosis kit
A technology for activating proteins and fibroblasts, which is used in medicine, the diagnosis and treatment of pancreatic cancer, and can solve problems such as no in-depth research.
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Embodiment 1
[0044] Example 1: Correlation between fibroblast activation protein in the stroma of pancreatic cancer and the clinicopathological characteristics and prognosis of pancreatic cancer
[0045] 1. Subjects
[0046] Pathological tissue samples were obtained from Peking Union Medical College Hospital, Chinese Academy of Medical Sciences. Sample inclusion and exclusion criteria are as follows:
[0047] (1) Inclusion criteria: cases undergoing surgical resection, and postoperative pathological diagnosis of pancreatic ductal adenocarcinoma;
[0048] (2) Exclusion criteria: incomplete case data, no pathology and follow-up data; tumors were not resected, or tumor biopsy was performed only, and complete histological specimens were not obtained; no corresponding paraffin specimens were found in the pathological specimen bank; paraffin specimens Sections for immunohistochemistry could not be prepared.
[0049] In this example, 56 pancreatic cancer patients were included, including 24 ma...
Embodiment 2
[0077] Example 2 Pancreatic cancer cell culture and passage
[0078] 1. Materials
[0079] (1) The human pancreatic cancer cell lines BxPC3, Capan1 and PANC1 were donated by Professor Freiss.H of the Technical University of Munich, Germany.
[0080] The surgically resected non-tumor pancreatic tissues of pancreatic cancer patients hospitalized for basic surgery in Peking Union Medical College Hospital were collected, and primary human pancreatic stellate cells were obtained, marked with PSC1#, PSC2#, ...PSCn#.
[0081] (2) Consumables, reagents and instruments related to cell culture
[0082] Reagent / consumable name
manufacturers
Cell culture flasks, cell freezing tubes, centrifuge tubes
Corning
Disposable pipette
Costar
COIC
Electric Auxiliary Pipette D03923
Accu-JetPro
High Pressure Steam Cooker MLS3750
SANYO
SANYO
Cell counter NeucleoCounter...
Embodiment 3
[0095] Example 3. Identification of primary human pancreatic stellate cells and expression of FAPα
[0096] 1. Identification of human primary pancreatic stellate cells and the expression of FAPα by immunofluorescence
[0097] (1) Relevant consumables, reagents and instruments
[0098] Table 4. Relevant consumables, reagents and instruments required for immunofluorescence detection
[0099]
[0100] (2) method
[0101] Human primary pancreatic stellate cells were planted on Nunc Lab-Tek Ⅱ chamber coverslips, respectively, with serum-free DMEM / F12 medium, DMEM / F12 medium + TGFβ1 (10ng / ml), pancreatic cancer cell BxPC3 , Capan1 and PANC1 conditioned medium culture, pancreatic cancer cells BxPC3, Capan1 and PANC1 conditioned medium culture + TGFβ neutralizing antibody, when the pancreatic stellate cells confluence reached 80-90%, perform immunofluorescence detection; remove the culture Base, add 100 μl formalin solution to each well, fix for 30 minutes; suck out the formali...
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