Fig 2: miR-6745 suppresses gastric tumor growth in vivo. Subcutaneous xenografts of GC cells infected with miR-6745 overexpressing lentivirus (Lv-miR-6745) or control lentivirus (Lv-miR-NC). (A) Images of the tumors at autopsy from nude mice are presented. (B) Tumor volumes and average weight of xenografted tumors were measured. (C) Immunohistochemical (IHC) staining of TIMP1 and Ki67 in xenografted tumors from Lv-miR-6745 cells or control cells. Data represent the means ± SEM. **P < 0.01.

Fig 3: SIRT6 protein expression is high in ovarian cancer, which regulates the metastatic phenotype (a) Immunohistochemical staining shows high SIRT6 levels in high-grade carcinoma patient tissue sections with corresponding bar diagram for expression levels. Scale bar = 10 µm. n = 3. Paired two-tailed t-test was done to calculate the P-value. (B) Human Protein Atlas data show high SIRT6 expression in ovarian cancer tissue. (C) qPCR data show elevated SIRT6 levels in PA1 cell lines compared with IOSE-364. n = 3, Paired two-tailed t-test was done to calculate the P-value. (D) Endogenous SIRT6 expression in IOSE-364 vs. PA1 and PA1 vs. SKOV3 cell lines. (E) Ki-67 staining in control (untransfected), EV-transfected, SIRT6-overexpressed, and SIRT6 siRNA transfected PA1 cells and relative fluorescence intensity have been depicted as bar graphs. Scale bar = 10 µm, n = 3 (F) qPCR data showed non-significant change in MMP2 and an increase in MMP9 in EV-transfected vs. pcDNA 3.1-SIRT6-transfected PA1 cells. n = 3, Paired two-tailed t-test was done to calculate the P-value. (G) Matrigel invasion studies show the number of invaded cells in IOSE-364, untransfected PA1 (control), EV-transfected, SIRT6-overexpressed, scrambled siRNA, and SIRT6-silenced PA1 cells. Corresponding bar diagram to show the number of invaded cells per field. Scale bar = 200 µm. n = 3, ANOVA was done to calculate the P-value. Error bars represent standard error of mean (SEM) from three independent experiments. ns—non-significant, *P-value < 0.05, **P-value < 0.01, ***P-value < 0.001. [Colour figure can be viewed at wileyonlinelibrary.com]

Fig 4: Recombinant scyreprocin (rScyreprocin) suppressed tumor growth in vivo. (A) Relative tumor volume (RTV) in different treatment groups. Cisplatin (CDDP, 3 mg kg-1), rScyreprocin (1.8 mg kg-1), and PBS (control) were given by intratumor injections; tumor volumes were measured every 3 days (n = 6). (B) Body weight of mice in different treatment groups. The body weights of the mice were measured every 3 days (n = 6). (C) Tumor tissues dissected from mice with different treatments. (D) Tumor weight of each group (n = 6). (E) H&E, TUNEL, Ki-67, and CD31 staining of tumor tissue sections obtained from mice with different treatments (scale bar = 50 µm). (F) Quantification of percent of Ki-67+ and CD31+ areas of each group (n = 5). Data are presented as means ± standard deviation (SD). In (A), data were normalized to the initial tumor volume measured at day 0. In (B), data were normalized to the control group and analyzed by one-way ANOVA with Tukey post-tests (*** p < 0.0001). In (F), data were analyzed by two-way ANOVA with Bonferroni post-test (*** p < 0.0001).

Fig 5: Tumor model viability over a 15-day culture period represented by ovarian cancer cell line xenografts (TOV21G). (a) Representative images of tumor models after 10 days in culture stained for nuclei (Dapi), human tumor cells (CK8/18+mito), cell proliferation (Ki67), and cell apoptosis (CC3). (b) IF scoring of tumor models showing stable expression of tumor-cell-specific marker (CK8/18+mito) over the 15-day period. (c–e) IF scoring of tumor models showing higher proliferation (c), larger nucleated area (d), and lower apoptotic cell death (e) in MDTs compared to tissue slices. Scale bars = 100 µm. Error bars = ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.0001; **** p < 0.00001.