Development of A New Mouse Model for Intrahepatic Cholangiocellular Carcinoma: Accelerating Functions of Pecam-1
Abstract
:1. Introduction
2. Results
2.1. Gross Anatomy and Microscopy of Liver in WT- and Pecam-1-KO-Mice after TAA-Administration
2.2. Immunohistological Identification of ICC in Murine Liver after TAA-Administration
2.3. Infiltration of Neutrophilic Granulocytes in Liver after TAA-Administration
2.4. Localization of Macrophages in Liver after TAA-Administration
2.5. Elevated Liver Markers in Serum of WT and Pecam-1 KO Mice after TAA Administration
2.6. Changes in Hepatic Gene Expression of Adhesion Molecules
2.7. Changes in Hepatic Gene Expression of Pro-Inflammatory Cytokines
2.8. Changes in Hepatic Gene Expression of Ccl5 and Lcn-2
2.9. Changes in Hepatic Gene Expression of c-Myc, Mmp-2, and Mmp9
2.10. Changes in Hepatic Gene Expression of EMT Markers
3. Discussion
3.1. Animal Models of Intrahepatic Cholangiocellular Carcinoma
3.2. Role of PECAM-1 in Inflammation-Induced Cancer
4. Material and Methods
4.1. Materials
4.2. Animals
4.3. Quantification of Hepatic Markers in Murine Serum
4.4. Immunofluorescence of Murine Liver Sections
4.5. RNA Isolation and Real-Time PCR Analysis
- Pecam-1: forward, AACAGAAACCCGTGGAGATG, reverse, GTCTCTGTGGCTCTCGTTCC;
- Icam-1: forward, ATTCGTTTCCGGAGAGTGTG, reverse, CAGCACCGTGAATGTGATCT;
- EpCAM: forward, GCTGTCATTGTGGTGGTGTC, reverse, TGGATCTCACCCATCTCCTT;
- Tgf-β: forward, TTGCTTCAAGCTCCACAGAGA, reverse, CAGAAGTTGGCATGGTAGCC;
- Tnf-α: forward, CAAACCACCAAGTGGAGGAG, reverse, GTGGGTGAGGAGCACGTAGT;
- Il-6: forward, CAAAGCCAGAGTCCTTCAGAG, reverse, GAGCATTGGAAATTGGGGTA;
- Ccl5/Lix: forward, GGTCCACAGTGCCCTACG, reverse, GCGAGTGCATTCCGCTTA;
- Lcn-2: forward, AAATTGCACAGGTATCCTCAG, reverse, CAGAGAAGATGATGTTGTCGT;
- C-Myc: forward, CGGACACACAACGTCTTGGAA, reverse, AGGATGTAGGCGGTGGCTTTT;
- Mmp-2: forward, GCTGTATTCCCGACCGTTG, reverse, TGGTCCGCGTAAAGTATGGG;
- Mmp-9: forward, ACGACATAGACGGCATCCAGTATC, reverse, AGGTATAGTGGCACACATAGTGGG;
- Wnt-5a: forward, AAGCAGGCCGTAGGACAGTA, reverse, CAGCACGTCTTGAGGCTACA;
- Twist-1: forward, GAGCAAGATTCAGACCCTCAA, reverse, CATCTTGGAGTCCAGCTCGT;
- B2m: forward, ATTCACCCCCACTGAGACTG, reverse, ATCCCAGTAGACGGTCTTGG;
- GAPDH: forward, AGAACATCATCCCTGCATCC, reverse, CACATTGGGGGTAGGAACAC.
4.6. Statistical Analysis
4.7. Data Availability
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Mouse | Liver Disease | Stage | Inflammation Grade |
---|---|---|---|
WT | Cirrhosis | 4 | 1–2 |
WT | Cirrhosis | 4 | 1–2 |
WT | Cirrhosis | 4 | 1–2 |
WT | Cirrhosis | 4 | 1–2 |
WT | Cirrhosis | 4 | 1–2 |
WT | Cirrhosis | 4 | 1–2 |
Pecam-1-KO | Cirrhosis | 3 | 1 |
Pecam-1-KO | Cirrhosis | 3 | 1 |
Pecam-1-KO | Cirrhosis | 3 | 1 |
Pecam-1-KO | Cirrhosis | 3 | 1 |
Pecam-1-KO | Cirrhosis | 3 | 1 |
Pecam-1-KO | Cirrhosis | 3 | 2 |
Antibody | Company | Cat. Number | IHC |
---|---|---|---|
rb-anti-ms-CK-19 | Abcam | ab 76539 | 0.180556 |
rt-anti-F4/80 (BM8) | Abcam | ab 16911 | 1:10 |
rt-anti-ms Ly-6G/ly-6C (Gr-1) | Invitrogen | 14-5931-81 | 0.180556 |
sh-anti-ms-CEACAM-1 | R &D systems | AF-6480 | 0.111111 |
ms-anti-SMA | Sigma | A-2547 | 0.180556 |
Secondary Antibodies: | |||
gt-anti-rt | |||
gt-anti-rb | Molecular Probes | --- | 0.180556 |
dk-anti-rb | |||
dk-anti-sh | Dianova | 713-225-147 | 0.180556 |
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Malik, I.A.; Malik, G.; Ströbel, P.; Wilting, J. Development of A New Mouse Model for Intrahepatic Cholangiocellular Carcinoma: Accelerating Functions of Pecam-1. Cancers 2019, 11, 1045. https://doi.org/10.3390/cancers11081045
Malik IA, Malik G, Ströbel P, Wilting J. Development of A New Mouse Model for Intrahepatic Cholangiocellular Carcinoma: Accelerating Functions of Pecam-1. Cancers. 2019; 11(8):1045. https://doi.org/10.3390/cancers11081045
Chicago/Turabian StyleMalik, Ihtzaz Ahmed, Gesa Malik, Philipp Ströbel, and Jörg Wilting. 2019. "Development of A New Mouse Model for Intrahepatic Cholangiocellular Carcinoma: Accelerating Functions of Pecam-1" Cancers 11, no. 8: 1045. https://doi.org/10.3390/cancers11081045