DOI: 10.5061/dryad.9p8cz8wt0
title: Data from: In vitro properties of patient serum predict clinical outcome after high dose rate brachytherapy of hepatocellular carcinoma

description: Tumor recurrence after local tumor ablation, including high-dose rate
 brachytherapy (HDR‑BT), represents a substantial challenge in
 hepatocellular carcinoma (HCC) treatment. This study aimed to investigate
 whether induced factors that appear in patient serum after HDR-BT alter
 HCC growth in vitro, and whether this correlates with outcome. In total,
 23 HCC patients [Barcelona clinic liver cancer (BCLC) stage A or B] were
 treated by HDR‑BT (1×15 Gy) and classified as responders in case of no
 progression within 6 months and no diffuse systemic progression within 2
 years (n=12), or non-responders with recurrence within 6 months and/or
 diffuse systemic tumor progression or extrahepatic disease within 2 years
 (n=11). Patient serum was obtained at baseline and 48 h post-procedure.
 Two hepatoma cell lines (HepG2, Huh7) were incubated for 72 h in the
 presence of 20% serum. BrdU incorporation was assessed for serum
 incubation at baseline and 48 h post-HDR‑BT. BrdU incorporation
 post-HDR‑BT compared to baseline was significantly elevated in
 non-responders compared to responders for both Huh7 and HepG2. Likewise,
 confirmatory Cyclin E studies revealed different induction kinetics
 between a subset of representative responders and non-responders in HepG2.
 Time to systemic progression (TTSP) in patients with increased BrdU
 incorporation was significantly shorter compared to patients with
 decreased BrdU incorporation after serum incubation. These data indicate
 that poor outcome following HDR‑BT is associated with increased measurable
 proliferation parameters of hepatoma cell lines in vitro after
 exposure to patient serum, offering insights into post-treatment tumor
 biology and a potential biomarker of clinical outcome.
description: # Data from: In vitro properties of patient serum predict clinical outcome
 after high dose rate brachytherapy of hepatocellular carcinoma
 [https://doi.org/10.5061/dryad.9p8cz8wt0](https://doi.org/10.5061/dryad.9p8cz8wt0) ## Description of the data and file structure Sheet 1 (‘BrdU incorporation HepG2, Huh7’): Shown are basic characteristics of the investigated patients as well as the BrdU incorporation of HepG2 and Huh7 in the serum incubation experiments. Briefly, serum of responders and non-responders to high dose rate brachytherapy (HDR-BT) was collected before (pre) and 48 h after (post) the procedure. Patient serum was used to incubate HepG2 and Huh7 cells in vitro for a period of 72 h. After incubation, the incorporation of BrdU (5-bromo-2'-deoxyuridine) was determined in HepG2 and Huh7 cells. The BrdU incorporation was evaluated as the BrdU positive area in relation to the DAPI positive area in the entire high-power field. For each cell line, the experiments were carried out twice (Experiment 1, Experiment 2). Each row represents one individual patient. Please find a detailed description of the column legends below. A: Patient ID in the manuscript B: Gender of the patient (male, female) C: Response of the patient to HDR-BT, please see ‘Material and Methods’ for a detailed description of the response evaluation D: Time to systemic progression (TTSP) indicating the period from study inclusion to systemic progression in days E – K represent data obtained from the experiments with HepG2. E – G represent the data from Experiment 1. H – J represent the data from Experiment 2. K represents the mean of values obtained in Experiment 1 and Experiment 2. K represents the mean of the two experiments. E: Experiment 1 with HepG2 cells: BrdU incorporation with patient serum collected before HDR-BT (pre) F: Experiment 1 with HepG2 cells: BrdU incorporation with patient serum collected after HDR-BT (post) G: Experiment 1 with HepG2 cells: post/pre ratio of the BrdU incorporation, values from column F (post) were divided by values from column E (pre) H: Experiment 2 with HepG2 cells: BrdU incorporation with patient serum collected before HDR-BT (pre) I: Experiment 2 with HepG2 cells: BrdU incorporation with patient serum collected after HDR-BT (post) J: Experiment 2 with HepG2 cells: post/pre ratio of the BrdU incorporation, values from column I (post) were divided by values from column H (pre) K: mean of the post/pre ratio from Experiment 1 (column G) and Experiment 2 (column J) L – R represent data obtained from the experiments with HepG2. L – N represent the data from Experiment 1. O – Q represent the data from Experiment 2. R represents the mean of values obtained in Experiment 1 and Experiment 2. R represents the mean of the two experiments. L: Experiment 1 with Huh7 cells: BrdU incorporation with patient serum collected before HDR-BT (pre) M: Experiment 1 with Huh7 cells: BrdU incorporation with patient serum collected after HDR-BT (post) N: Experiment 1 with Huh7 cells: post/pre ratio of the BrdU incorporation, values from column M (post) were divided by values from column L (pre) O: Experiment 2 with Huh7 cells: BrdU incorporation with patient serum collected before HDR-BT (pre) P: Experiment 2 with Huh7 cells: BrdU incorporation with patient serum collected after HDR-BT (post) Q: Experiment 2 with Huh7 cells: post/pre ratio of the BrdU incorporation, values from column P (post) were divided by values from column O (pre) R: mean of the post/pre ratio from Experiment 1 (column N) and Experiment 2 (column Q)   Sheet 2 (‘Cyclin E’): Shown are the Cyclin E expression levels in HepG2 after cell cycle induction with serum obtained before and after HDR-BT. Columns represent data of different serum incubation periods (baseline/ 0 h, 3 h, 6 h, 9 h, 11 h, 13 h). The Cyclin E expression was evaluated as the Cyclin E positive area in relation to the DAPI positive area in the entire high-power field. Each row represents one individual patient. Please find a detailed description of the column legends below. A: Patient ID of three representative responders and non-responders B: Cyclin E expression after incubation for a period of 0h with serum obtained before HDR-BT. C: Cyclin E expression after incubation for a period of 0h with serum obtained after HDR-BT. D: post/pre ratio of Cyclin E expression with an incubation period of 0h, values from column C divided by values from column B E: Cyclin E expression after incubation for a period of 3h with serum obtained before HDR-BT. F: Cyclin E expression after incubation for a period of 3h with serum obtained after HDR-BT. G: post/pre ratio of Cyclin E expression with an incubation period of 3h, values from column F divided by values from column E H: Cyclin E expression after incubation for a period of 6h with serum obtained before HDR-BT. I: Cyclin E expression after incubation for a period of 6h with serum obtained after HDR-BT. J: post/pre ratio of Cyclin E expression with an incubation period of 6h, values from column I divided by values from column H K: Cyclin E expression after incubation for a period of 9h with serum obtained before HDR-BT. L: Cyclin E expression after incubation for a period of 9h with serum obtained after HDR-BT. M: post/pre ratio of Cyclin E expression with an incubation period of 9h, values from column L divided by values from column K N: Cyclin E expression after incubation for a period of 11h with serum obtained before HDR-BT. O: Cyclin E expression after incubation for a period of 11h with serum obtained after HDR-BT. P: post/pre ratio of Cyclin E expression with an incubation period of 11h, values from column O divided by values from column N Q: Cyclin E expression after incubation for a period of 13h with serum obtained before HDR-BT. R: Cyclin E expression after incubation for a period of 13h with serum obtained after HDR-BT. S: post/pre ratio of Cyclin E expression with an incubation period of 13h, values from column R divided by values from column Q   Sheet 3 (‘FCS deprivation’): Shown is the BrdU incorporation of Huh7 and HepG2 cells after incubation in medium that was either fetal calf serum (FCS)-supplemented or FCS-deprived. The BrdU incorporation was evaluated as the BrdU positive area in relation to the DAPI positive area in the entire high-power field. Each row represents one experiment. Please find a detailed description of the column legends below. A: Number of experiment B: BrdU incorporation of Huh7 cells after incubation with FCS-supplemented medium C: BrdU incorporation of Huh7 cells after incubation with FCS-deprived medium D: BrdU incorporation of HepG2 cells after incubation with FCS-supplemented medium E: BrdU incorporation of HepG2 cells after incubation with FCS-deprived medium   Sheet 4 (‘PTN, CRTAM’): Shown are post/pre ratios of BrdU incorporation of HepG2 and Huh7 cells after incubation with patient serum for 72 h as well as post/pre ratios of plasma levels of proteins Pleiotrophin (PTN) and Cytotoxic and regulatory T Cell Molecule (CRTAM). The protein plasma levels were detected by Proximity-extension assay and are reported in arbitrary units. Each row represents one individual patient. Please find a detailed description of the column legends below. A: Patient ID in the manuscript B: post/pre ratio of BrdU incorporation of HepG2 cells C: post/pre ratio of BrdU incorporation of Huh7 cells D: post/pre ratio of plasma levels of PTN E: post/pre ratio of plasma levels of CRTAM ## Human subjects data The authors confirm that all analyses were performed on anonymized data. To further ensure de-identification, variables such as age and other potentially identifying informatino were removed from the provided dataset.