FIGURE 7.

Therapeutic MNVs alter the tumor microenvironment. Livers were harvested, and serum samples were collected from orthotopic HCC tumor-bearing mice treated with control MNVs or tMNVs and adoptively transferred NK cells. (A) RNA was extracted, and miR-126-3p and U6 expression was assessed by RT-PCR. (B, C). Expression of ADAM9 in protein lysates. Representative immunoblots (B) and quantitative data (C) from 4 samples are shown. (D) Soluble MICA levels were quantitated in serum samples by ELISA. (E) VEGF level in tumor tissues was quantified by ELISA. (F) H&E and immunohistochemistry for CD56 were performed in sections from tumor and nontumor regions of the liver. The arrows indicate cells that express CD56. (G) Quantitation of CD56 positivity within cells in tumoral and non-tumoral regions. (H) Serum cytokine and chemokine levels. The values are expressed as the mean andSD (n = 4 mice per group). p < 0.05, ***p<0.001. Scale bars represent 200 µm. Abbreviations: H&E, Hematoxylin and Eosin; MICA, major histocompatibility complex I-related chain A; MNVs, milk-derived nanovesicles; NK cells, natural killer cells; tMNVs, therapeutic MNVs.