| Objective: Observe the expression of PD-L1 molecule in the tissues of patients with intrahepatic cholangiocarcinoma.Observe and analyze the expression of PD-L1,IFN-?,TNF-? and VEGF in ICC tissue.Observe the therapeutic effect of PD-L1 antibody on mouse ICC in situ model.Methods: The cancer tissues of 60 patients with ICC,who were selected and treated in the First Affiliated Hospital of Bengbu Medical College from January 2014 to December 2017 and 40 of them were selected as controls.And the expression of PD-L1 molecules in tissues adjacent to cancer,and analyze the relationship between PD-L1 expression and clinicopathological factors.Put p T3-myr-AKT-HA,p T3-EF1a-NICD1 and C-luc-SB13 dissolved in 2m L of 0.9% Na Cl solution,and the0.9% Na Cl solution containing these three plasmids is injected into mice within 7seconds by using hydrodynamic methods to establish an in-situ model of mouse ICC.The animal imaging system and animal B-ultrasound was used to observe the results of the tumor model construction,and the ICC tumor tissues of tumor-bearing mice were stained with HE to prove their modeling status.Normal liver tissues and ICC tumor tissues of tumor-bearing mice were taken to detect concentration of PD-L1 and tumor microenvironment-related factors IFN-?,TNF-?,VEGF,using Pearson correlation coefficient method to analyze the correlation between PD-L1 and IFN-?,TNF-?,VEGF.ICC mice were given PD-L1 antibody treatment,through animals Imager and B-ultrasound of animals to observe the therapeutic effect of PD-L1 antibody.Results: The positive rate of PD-L1 molecules in ICC tissues is 51.7%,and the positive rate in adjacent tissues is 5%.The expression of PD-L1 molecules in ICC is significantly higher than that in adjacent tissues,the difference is statistically significant(51.7% vs 5%,P<0.05).PD-L1 expression has nothing to do with the patient's gender,age,tumor diameter and degree of differentiation,and the difference is not statistically significant(P>0.05).PD-L1 expression is related to lymph node metastasis and TNM stage in ICC patients and the difference is statistically significant(P<0.05).After 4 weeks,all mice had tumors,with a tumor formation rate of 100%.The liver of the tumor-bearing mice showed strong bioluminescence when observed by the small animal imager.Observed by B-ultrasound of small animals,liquid dark areas and tumor echo can be seen in the liver of tumor-bearing mice.HE staining results showed that the tumor cells in the tumor tissue section of the tumor-bearing mice were arranged like a bile duct.ELISA results showed that PD-L1 molecule was highly expressed in ICC mice,and its expression had a linear relationship with tumor microenvironment-related factors IFN-?,TNF-? and VEGF.Detected the tumor-bearing mice that were treated with PD-L1 antibody by using animal imaging system,and the bioluminescence signal in the liver was weakened.The observation of the small animal B-ultrasound,after the tumor-bearing mice were treated with PD-L1 antibody,the liquid dark areas in the liver area shrank.Conclusion: The PD-L1 molecule is highly expressed in ICC tissues,but is low or not expressed in adjacent tissues.It is expression is not related to the patient's gender,age,tumor diameter and degree of differentiation,and is related to the lymph node metastasis and TNM stage of ICC patients.The high expression of PD-L1 molecule is related to ICC tumor microenvironment cytokines.PD-L1 antibody has a therapeutic effect on the mouse ICC in situ model. |
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