Deficiency of the promyelocytic leukemia protein fosters hepatitis C-associated hepatocarcinogenesis in mice.

Overwhelming lines of epidemiological evidence have indicated that persistent infection with hepatitis C virus (HCV) is a major risk for the development of hepatocellular carcinoma (HCC). We have recently shown that HCV core protein mediates functional inactivation of the promyelocytic leukemia (PML) tumor suppressor pathway. However, the role of PML in HCC development yet remains unclear. To clarify the function of PML in liver carcinogenesis and HCV-associated pathogenesis we crossed PML-deficient mice with HCV transgene (HCV-Tg) expressing mice and treated the resulting animals with DEN/Phenobarbital, an established protocol for liver carcinogenesis. Seven months after treatment, livers were examined macroscopically and histologically. Genetic depletion of the tumor suppressor PML coincided with an increase in hepatocyte proliferation, resulting in development of multiple dysplastic nodules in 100% of the PML-deficient livers and of HCCs in 53%, establishing a tumor suppressive function of PML in the liver. In animals expressing the HCV-transgene in PML-deficient background, HCC development occurred even in 73%, while only 7% of their wildtype littermates developed HCC. The neoplastic nature of the tumors was confirmed by histology and expression of the HCC marker glutamine synthetase. Several pro- and antiapoptotic factors were tested for differential expression and liver carcinogenesis was associated with impaired expression of the proapoptotic molecule TRAIL in PML-deficient mice. In conclusion, this study provides first in vivo evidence that the tumor suppressor PML acts as an important barrier in liver carcinogenesis and HCV-dependent liver pathology

Additional file 1 of Highly sensitive therapeutic drug monitoring of infliximab in serum by targeted mass spectrometry in comparison to ELISA data

Supplementary Material 1: Calibration curves of the ASQ peptide of infliximab. Curves were generated and measured by two different researchers with a time difference of 6 month. Both calibration curves show robust results in regard to their LOD and LLOQ. Each calibration point was measured 3 time with independent replicates. The LOD is marked with the purple line, the LLOQ is marked with the blue lin

PML-expression influences liver weight.

<p>(A) Body weight, (B) liver weight as well as (C) liver/body weight ration of all mouse strains, either treated or not, was determined.</p

Characterisation of PML-deficient HCV-transgenic mice.

<p>(A) Whole liver extracts derived from 4 weeks old wild-type and HCV-PML+ mice were analysed by immunoblot analysis for the protein expression of PML and HCV-core as well as tubulin (loading control). (B) Whole liver extracts from 4 weeks old PML-HCV+ mice as well as WT mice were analysed for the mRNA expression levels of PML and HCV core by quantitative RT-PCR.</p

Expression of pro- and antiapoptotic genes.

<p>(A) mRNA expression analysis of several anti-apoptotic factors (Mcl-1, Survivin, Bcl-2, XIAP, Bcl-xL) by quantitative RT-PCR in the four treated transgenic groups. Boxes represent the average expression level of control. All values are normalized to SDHA mRNA expression. Standard deviation is indicated by error bars. (B) mRNA expression analysis of TRAIL by quantitative RT-PCR, comparing the four treated mouse strains versus untreated controls. Boxes represent the average expression level of control. All values are normalized to SDHA mRNA expression. Standard deviation is indicated by error bars. (C) Whole liver extracts derived from 7 months old mice were analysed by immunoblot analysis for the protein expression of TRAIL as well as tubulin (loading control). A representative of five independent experiments is shown.</p