Effect of <i>MIC-1/GDF15</i> gene modification on metastases.

<p>(A) Survival data for TRAMP^MIC+/+ (n = 63), TRAMP^MIC-/- (n = 63) and TRAMP^fmsmic-1 (n = 63) mice is presented as a Kaplan-Meier plot and the log-rank statistic for median survival time is shown. (B) Comparison between number of TRAMP^MIC+/+ (n = 63), TRAMP^MIC-/- (n = 63) and TRAMP^fmsmic-1 (n = 63) mice having distant organ metastases at the time of death has been analyzed using the Chi-squared test.</p

TRAMP^MIC-/- mice have comparatively larger prostate tumor than TRAMP^MIC+/+ mice.

<p>The corrected tumor weights of (A) GU and (B) prostates were compared in TRAMP^MIC+/+ and TRAMP^MIC-/- mice (n = 22/group/time point) sacrificed at 8, 17, 25 and 33 weeks of age. Results were analyzed using a 2 way ANOVA and are presented as mean ± SEM. p values are shown as *, p<i><</i> 0.05.</p

TRAMP^MIC-/- mice have shorter survival and larger prostate tumors than TRAMP^MIC+/+ mice.

<p>(A) Survival data for TRAMP^MIC+/+ and TRAMP^MIC-/- mice (n = 35). Overall survival of individual mice from birth to death was plotted using the Kaplan-Meier method. The log-rank statistic for median survival time is shown. (B) The genitourinary complex (GU) and prostate tumor weights (C), in TRAMP^MIC+/+ and TRAMP^MIC-/- mice, at the necropsy, are corrected for body weight and presented as mean ± SEM. Differences are analyzed using an unpaired 2-tailed t test. (D) The number of TRAMP^MIC+/+ and TRAMP^MIC-/- mice having large prostate tumor (corrected tumor wt>10mg/g), was compared using a Chi-square test. p values are shown as *, p<i><</i> 0.05; **, p<i><</i> 0.01.</p

TRAMP^fmsmic-1 mice survive longer and have smaller prostate tumors than TRAMP mice.

<p>Overall survival of individual mice from birth to death is shown (panel A). The survival data for TRAMP (—▴—) and TRAMP^fmsmic-1 (—Δ—) mice were plotted using the Kaplan-Meier method. The log-rank statistic for median survival time is shown. The genitourinary (GU), seminal vesicle (SV) and prostate tumor sizes in TRAMP^fmsmic-1 and TRAMP mice (n = 15/group) sacrificed at 8, 17, 25 and 33 weeks of age are shown in panel B, C and D. The GU (panel B), SV (panel C) and prostate weights (panel D) are presented as mean mg weight/g body weight ± SEM. p values are shown as *p<i><</i>0.05; **p<i><</i>0.01; ***p<i><</i>0.001.</p

TRAMP^fmsmic-1 prostate tumors have lower histological grades.

<p>Prostates of 15 TRAMP and 15 TRAMP^fmsmic-1 mice were excised at week 8 (panel A), 17 (panel B), 25 (panel C), and 33 (panel D). Serial sections were graded and scores for each grade were averaged for all the mice in the group. The graph represents the mean proportion of the prostate from all the mice in the group having each different pathologic grade, scored between 1 and 6± SEM. Numbers on the bars show number of mice in the group having a particular grade. p values are shown as *p<i><</i>0.05; **p<i><</i>0.01; ***p<i><</i>0.001.</p

MIC-1/GDF15 increases metastasis.

<p>(A) Comparison between proportion of TRAMP (n = 30) and TRAMP^fmsmic-1 (n = 30) mice having distant organ metastasis. The data is analyzed by Chi-square test. (B) Comparison between proportion of TRAMP (n = 59) and TRAMP^fmsmic-1 (n = 33) mice with metastasis at 18–40 weeks, a period where TRAMP and TRAMP^fmsmic-1 mice reach ethical end points at the same rate. The data is analyzed by Chi-square test. (C) Comparison between number of lung tumor colonies in MIC-1^fms mice and control C57BL/6 mice injected intravenously with TC1-T5 cells. The data is analyzed by Unpaired <i>t</i> test. The p values for Chi-square test and Unpaired <i>t</i> test are shown as ***p<i><</i>0.001.</p

<i>MIC-1/GDF15</i> expression in TRAMP and TRAMP^fmsmic-1 prostate and TC1-T5 cell line.

<p>(A) <i>MIC-1/GDF15</i> expression was quantified by qRT-PCR in the prostate of TRAMP and TRAMP^fmsmic-1 mice at 8, 17 and 25 weeks and normalized to <i>B-actin</i> expression as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0043833#s2" target="_blank">material and methods</a>. (B) Relative <i>MIC-1/GDF15</i> expression was quantified by qRT-PCR in the TC1-T5 cell line (n = 3) and compared with TRAMP (n = 3) and TRAMP^fmsmic-1 (n = 3) prostate tumor after normalizing with <i>TBP</i> expression. Values are presented as mean normalized <i>MIC-1/GDF15</i> expression ±SEM. p values for the two-tailed unpaired <i>t</i> test are shown as ***p<i><</i>0.001.</p