Active Forms of Chemerin Are Elevated in Human and Mouse Ovarian Carcinoma

Figures

• 

  Figure 1. Changes in levels of chemerin forms in plasma and serum from patients with BMI < 25 and patients with BMI > 40. Blood was obtained, and either plasma or serum was used prior to measurement of chemerin by ELISA. (a) levels of different chemerin forms in plasma and serum from volunteers < 25 BMI (N = 22) (b) levels of different chemerin forms in plasma and serum from people with obesity with BMI > 40 (N = 22). P: plasma, S: serum.

• 

  Figure 2. Changes in the levels of chemerin forms in the plasma and serum from two cohorts, BMI < 25 and BMI > 40 (each group N = 22), as determined by ELISA. Blood was obtained and either plasma or serum was prepared before measuring levels of chemerin by ELISA. P: plasma, S: serum. (a) levels of different chemerin forms in plasma and serum from volunteers with BMI < 25. Total chemerin, chem163S, and cleaved chemerin plotted on the left Y axis, chem158K, checm157S, chem156F, chem155A, and degraded chemerin plotted on the right Y axis. (b) levels of different chemerin forms in plasma and serum from people with obesity (BMI > 40). Total chemerin, chem163S, and cleaved chemerin plotted on left Y axis, chem158K, checm157S, chem156F, chem155A, and degraded chemerin plotted on right Y axis. (c) Average % of different chemerin forms in plasma and serum from volunteers and people with obesity. Data was analyzed by two-tailed paired Student t test between plasma and serum samples. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

• 

  Figure 3. Inclusion of protease inhibitors does not significantly affect levels of chemerin forms in ascitic fluid samples. Samples from ascitic fluid of three ovarian carcinoma patients were processed with (+) or without (−) the addition of protease inhibitors before measuring levels of chemerin by ELISA.

• 

  Figure 4. Levels of total chemerin (total chem) in ascites from patients with ovarian carcinoma determined by ELISA (N = 19) (a) total chemerin/mL (b) total chemerin normalized to protein content of ascitic fluid. Samples marked as Serous and then by site of origin, Fallopian (Fallopian tube serous carcinoma), Ovarian (Serous ovarian carcinoma), Peritoneal (Primary peritoneal serous carcinoma) and other marked as C (clear cell ovarian carcinoma), D (de-differentiated ovarian carcinoma), and M (mucinous borderline ovarian carcinoma). Also included are four ascites samples from three patients with other cancers, G (gastric signet adenocarcinoma), S (cervical squamous adenocarcinoma), and U (uterine carcinosarcoma), and one benign, B (benign serous cyst, para-ovarian).

• 

  Figure 5. Levels of different chemerin forms in ascites from patients with ovarian carcinoma (a) chem163S (N = 16) (b) chem158K (N = 19) (c) chem157S (N = 19) (d) chem156F (N = 19) (e) chem155A (N = 16). Samples marked as Serous and then by site of origin, Fallopian (Fallopian tube serous carcinoma), Ovarian (Serous ovarian carcinoma), Peritoneal (Primary peritoneal serous carcinoma) and other marked as C (clear cell ovarian carcinoma), D (de-differentiated ovarian carcinoma), and M (mucinous borderline ovarian carcinoma). Also included are three ascites samples from patients with other cancers, G (gastric signet adenocarcinoma), S (cervical squamous adenocarcinoma), and U (uterine carcinosarcoma).

• 

  Figure 6. Levels of cleaved and degraded chemerin forms in ascites from patients with ovarian carcinoma (N = 16) (a) cleaved chemerin calculated as total chemerin–chem163S (b) degraded chemerin calculated as total chemerin–(chem163S + chem158K + chem157S + chem156F + chem155A). Samples marked as Serous and then by site of origin, Fallopian (Fallopian tube serous carcinoma), Ovarian (Serous ovarian carcinoma), Peritoneal (Primary peritoneal serous carcinoma), and one other marked as M (mucinous borderline ovarian carcinoma). Also included are three ascites samples from patients with other cancers, G (gastric signet adenocarcinoma), S (cervical squamous adenocarcinoma), and U (uterine carcinosarcoma).

• 

  Figure 7. Levels of different chemerin forms in ascites from patients with ovarian carcinoma determined by ELISA for total chemerin and specific forms. (a) Values of different forms for overall cohort. Data shown in a violin plot with thick dotted line representing the median and the thin dotted lines quartiles. N = 16 except for total chemerin where N = 22 and chem158K, chem157S, and chem156F where N = 19. (b) The % distribution for each specific chemerin form in patients for which complete ELISA data is available (N = 16) is displayed based on data in Figure 4a and Figure 5. The average % distribution is shown in the right-most column. Samples marked as Serous and then by site of origin, Fallopian (Fallopian tube serous carcinoma), Ovarian (Serous ovarian carcinoma), Peritoneal (Primary peritoneal serous carcinoma), and one other marked as M (mucinous borderline ovarian carcinoma). Also included are three ascites samples from patients with other cancers, G (gastric signet adenocarcinoma), S (cervical squamous adenocarcinoma), and U (uterine carcinosarcoma).

• 

  Figure 8. (a) Average levels of different forms of mouse chemerin in the ascites harvested from mice (n = 11) with ID8 tumors sampled at sacrifice. Total chemerin and the different chemerin forms were determined by ELISA. Levels of cleaved chemerin were calculated as total chemerin–chem163S, and degraded chemerin calculated as total chemerin–(mProchem + mchem157R + mchem156S + mchem155F + mchem154A). Data shown in a violin plot with thick dotted line representing the median and the thin dotted lines quartiles. (b) Levels of different specific chemerin forms in individual mice; shown as % based on data shown in Figure 7a. The average % distribution is shown in the right-most column.

• 

  Figure 9. WT and chemerin KO mice were monitored by weight after inoculation with ID8 tumor cells and control mice: WT con and chemerin KO con were injected with PBS. Mice were sacrificed after reaching the allowable maximum weight gain or on Day 84. (a) % Weight gain during the survival study. The black arrow represents the endpoint at which mice began to be sacrificed. WT and chemerin KO n = 8 (b) Kaplan–Meier curve of the survival of WT (n = 24), chemerin KO (chem KO; n = 21) mice, WT con (n = 8), and chemerin KO con (n = 8) mice. (c) Area under the curve (AUC) of weight gain shown as mean ± SEM from the data in (b) as described in Materials and Methods, Section 2.6. Data was analyzed by ANOVA followed by post hoc Tukey-Kramer test. * p < 0.05, ** p < 0.01, *** p < 0.001.