Perindopril sensitizes hepatocellular carcinoma to chemotherapy: A possible role of leptin / Wnt/ β-catenin axis with subsequent inhibition of liver cancer stem cells

Figures

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  Fig. 1. Experimental design. Sixty male mice were divided into six groups and treated with (DENA, DENA + Perindopril, DENA + sorafenib, DENA + Perindopril + sorafenib) as explained in the figure. More details in the text.

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  Fig. 2. Hepatoprotective effect of sorafenib/ Perindopril regimen: effect of drugs on ALT (Fig. 2 A) and AST (Fig. 2 B). Data are expressed as means ± SD. * Value significantly (p ≤ 0.001) different between groups. n = 10.

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  Fig. 3. Impact of sorafenib/ Perindopril regimen on metastasis, angiogenesis, and cell cycle progression. Fig. 3 illustrates the effect of sorafenib, Perindopril, and combination of them on the metastatic marker (MMP-9) (Fig. 3 A), angiogenic marker (VEGF) (Fig. 3B), and marker of cell cycle progression (cyclin D1) (Fig. 3 C) Data are expressed as means ± SD. * Value significantly (p ≤ 0.001) different between groups. n = 10.

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  Fig. 4. leptin / Wnt / β-catenin axis as a molecular target of sorafenib/ Perindopril regimen in DENA induced HCC.Fig. 4 illustrates the effect of sorafenib, Perindopril, and combination of them on Leptin (Fig. 4A), β catenin (Fig. 4B) and wnt expression (Fig. 4 C). Data are expressed as means ± SD. * Value significantly (p ≤ 0.001) different between groups. n = 10.

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  Fig. 5. Perindopril downregulates EpCAM expression.Fig. 5 illustrates the effect of sorafenib, Perindopril, and combination of them on EpCAM (Fig. 5 A) and ALDH (Fig. 5B). Data are expressed as means ± SD. * Value significantly (p ≤ 0.001) different between groups. n = 10.

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  Fig. 6. Gross pathological examination of livers: Livers from mice in each group were examined macroscopically. liver from control mice (Fig. 6A). Livers from DENA treated mice (Fig. 6B&C). Livers from mice treated with perindopril, sorafenib, or combination of both (Fig. 6D, E&F).

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  Fig. 7. Histopathological examination of liver tissues: Liver tissues were processed and stained with hematoxylin and eosin. Control as well as sorafenib and perindopril treated mice (without DENA) showed normal hepatic architecture (Fig. 6A-B). On the other hand, liver sections from DENA treated mice displayed multiple nodules in liver tissue. Each nodule showed pleomorphic hepatocytes arranged in trabeculae (more than two cells thick. Focal spindling could be seen. (Fig. 6C-D). The effect of sorafenib on liver tissue is represented in (Fig. 6E) where liver sections showed macrovesicular steatosis (mild steatosis) with a focal small area of hepatocyte degeneration. Preventative liver section from perindopril treated mice showed preserved normal liver architecture with moderate infiltration by lymphocytes in some portal tracts and nuclear dysplasia in the form of hyperchromatic nuclei (Fig. 6F). Liver sections from mice treated with combination therapy Sorafenib and Perindopril showed normal liver architecture with some lymphocytes infiltration in portal tract and interface hepatitis but no dysplastic changes were seen (Fig. 6G-H). The images shown are representative photomicrographs.…

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  Fig. 8. Schematic diagram of molecular targets of sorafenib and perindopril.sorafenib and perindopril inhibit Wnt/ β-catenin pathway whereas perindopril potentiates this inhibition through dampening leptin level. Leptin is associated with Akt phosphorylation and inactivation of GSK 3β (more details in text).