Evaluation of Phase I tumor volume values.

<p>“Measured data” is the MRI-measured tumor volume—tumor mass pairs on the 23rd day of Phase III/3 (case and control group). For this dataset, linear curve fitting was carried out (“fitted linear curve”) to find the mathematical relationship between MRI-measured tumor volume and tumor mass. Substituting tumor mass values which were measured on the 24th day of Phase I to the equation of the resulted curve, the corresponding tumor volume values can be evaluated (“evaluated data”).</p

Experimental settings for small animal MRI measurement.

<p>In the first four measurement times 30 slices, in the last two measurement times 40 slices were done from each mouse. Abbreviations: T1WI: T1 weighted image, FSE: Fast Spin Echo, FOV: Field of View [<i>mm</i>²], TR: Repetition Time, ESP: Echo Spacing.</p><p>Experimental settings for small animal MRI measurement.</p

Illustration for tumor with irregular structure (berry-shaped).

<p>a) berry-shaped tumor; b) <i>x</i>-diameter of the tumor; c) <i>y</i>-diameter of the tumor; d) <i>z</i>-diameter of the tumor; e) berry-shaped tumor with ellipsoidal estimation. Even though all the three diameters can be measured, the estimation of the volume has quite a large error.</p

Stained slices in the case of <i>n1</i> mouse (Phase I, 24th day of the experiment).

<p>a) Haematoxylin Eosin (H&E) staining was applied to investigate tumor morphology. b) Fluorescence picture was created using CD31 antibody immunohistochemistry staining to calculate vascularization area.</p

Experimental data of human colorectal adenocarcinoma (HT-29) xenografts (tumor mass).

<p>Experimental data of human colorectal adenocarcinoma (HT-29) xenografts (tumor mass).</p

Experimental data of C38 mouse colon adenocarcinoma (tumor length, tumor width, tumor mass and tumor volume).

<p>Data was measured at the final day of Phase I (24th day) and Phase III/3 (23rd day).</p><p>^a Directly measured data (tumor length, tumor width, tumor mass)</p><p>^b Estimated data (tumor volume measured by caliper, calculated according to Xenograft tumor model protocol or two-dimensional mathematical model)</p><p>^c MRI-measured data (tumor volume calculated with flood fill algorithm)</p><p>^d Evaluated data (“MRI” tumor volume calculated from linear curve fit (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0142190#pone.0142190.e008" target="_blank">Eq 8</a>))</p><p>Experimental data of C38 mouse colon adenocarcinoma (tumor length, tumor width, tumor mass and tumor volume).</p

Overview of Phase I and Phase III/3.

<p>In Phase I, tumor growth was investigated without antiangiogenic therapy for 24 days (mice were sacrificed when the tumor reached a lethal size). In Phase III/3, control group members received one 200 <i>μg</i> bevacizumab dose for an 18-day therapy (on the 3rd and 21st days); case group members received 1.11 <i>μg</i> bevacizumab every day for 20 days.</p

Validation of caliper-measured data.

<p>The figure shows the results of a mouse (<i>C4</i>) from control group (first row), and a mouse (<i>E9</i>) from case group (second row). The first column shows the tumor values which were calculated using the two-dimensional mathematical model; the second column represents the protocol-based tumor volumes. In each case the reference value is the MRI-measured tumor volume. One can see that the two-dimensional mathematical model fits to the MRI-measured values, while the protocol-based values present totally different curve.</p

Overview of Phase IV.

<p>In Phase IV, control group members received no bevacizumab. Case1 group members received one 200 <i>μg</i> bevacizumab dose on the 8th day, case2 group members received 1.11 <i>μg</i> bevacizumab from the 8th day every day for 15 days.</p

Additional file 2: Table S2. of EZH2 is a sensitive marker of malignancy in salivary gland tumors

Benign tumors. (DOCX 13 kb