[PMC free article] [PubMed] [Google Scholar] 22
June 3, 2021
[PMC free article] [PubMed] [Google Scholar] 22. the malignant ascites of ovarian cancer and promote cancer growth. iPS\ML, macrophage\like myelomonocytic cells generated from human induced pluripotent stem (iPS) cells, made close contacts Rabbit Polyclonal to PRKAG2 with ovarian cancer cells in?vitro. We hypothesized that, if we MitoTam iodide, hydriodide inoculate iPS\ML\producing IFN\ (iPS\ML/IFN\) into the peritoneal cavity of patients with ovarian cancer, IFN\ produced by the iPS\ML/IFN\ would efficiently act around the cancer cells to suppress cancer growth. To evaluate this hypothesis, we injected iPS\ML/IFN\ into SCID mice bearing peritoneally disseminated human ovarian cancer cells, SKOV3. Immunohistochemical analysis of the intraperitoneal tumors detected iPS\ML/IFN\ infiltrating into the MitoTam iodide, hydriodide cancer tissues. Therapy with iPS\ML/IFN\ significantly suppressed tumor progression. In addition, dramatic reduction of cancer\related ascites was observed. Collectively, it is suggested that iPS\ML/IFN\ therapy offers a new approach for the treatment of patients with advanced ovarian cancer. Jcl female mice were purchased from CLEA Japan. Mice were intraperitoneally (i.p.) injected with SKOV3 cells (2??107?cells/mouse) expressing luciferase. After 3 or 4 4?days, the mice were anesthetized by i.p. injection of medetomidine, midazolam, and butorphanol. The mice underwent bioluminescence imaging to examine the extent of ovarian cancer metastasis (NightOWL II; Berthold Technologies, Bad Wildbad, Germany). After confirmation of the engraftment of the cancer, mice were divided into control and treatment groups. The mice in the treatment group were injected twice each week with iPS\ML (without production of IFN\, 1??107?cells/mouse) or iPS\ML/IFN\ (1??107?cells/mouse). All mice underwent luminescence image analysis to evaluate the effects of the treatment. The experimental data were analyzed using Indigo analysis software. 2.8. Statistical analysis Statistical analyses were carried out using Student’s test (SPSS version 24, IBM; Armonk, NY, USA). A test) The effect of coculture with iPS\ML/IFN\ on the number of live SKOV3 and ES2 cells was also examined. We cocultured iPS\ML/IFN\ and the cancer cells expressing firefly luciferase. The number of live SKOV3 and ES2 cells was measured based on the luciferase activity after a 3\day culture. iPS\ML/IFN\ reduced the number of live SKOV3 and ES2 cells in a dose\dependent way (Physique?2). Coculture of ordinary\type iPS\ML (without production of IFN\) did not affect the number of SKOV3 cells in the absence or MitoTam iodide, hydriodide presence of recombinant IFN\ (Physique?S1). According to these findings, it is confirmed that iPS\ML had no direct anticancer effect. Open in a separate window Physique 2 Sensitivity MitoTam iodide, hydriodide of ovarian cancer cell lines to induced pluripotent stem\cell\derived myelomonocytic cells (iPS\ML)/interferon (IFN)\ in?vitro. Luciferase\expressing A, SKOV3 or B, ES2 cells were cultured in a 96\well culture plate (1??103?cells/well) with or without iPS\ML/IFN\. Number of live cancer cells was measured by luciferase activity after 3?days. The difference from the control was statistically significant (*test. RLU, relative luminescent models) 3.2. Cognate conversation of tumor cells and macrophages Direct conversation between macrophages and cancer cells plays a pivotal role in tumor progression. We previously reported the presence of abundant numbers of macrophages (106?cells/mL on average) in the ascites of patients with advanced stages of ovarian cancer, and the promotion of ovarian cancer cell growth by the conversation between macrophages and cancer cells. 5 A similar phenomenon was observed in this study, and most of the cells formed aggregates in the ascites of patients with ovarian cancer (Physique?3A). In addition to cancer cells, sediments of the ascites included a large number of CD68+?CD163+ macrophages (Physique?3B). We cocultured SKOV3 cancer cells with iPS\ML/IFN\, and the cells were fixed and stained with anti\CD68 antibody to distinguish iPS\ML/IFN\ from the malignancy cells. As shown in Physique?3C, iPS\ML/IFN\ were in close contact with SKOV3 cancer cells. Open in a separate window Physique 3 Conversation of macrophages with ovarian cancer cells. A, Spheres present in the ascites of serous carcinoma of the ovary (400). B,C, Presence of CD68\ and CD163\positive cells in precipitates of ascites of ovarian carcinoma was examined (200). D, Induced pluripotent stem\cell\derived myelomonocytic cells (iPS\ML)/interferon (IFN)\ were evaluated immunohistochemically using an anti\CD68 antibody (400). Distinct staining for CD68 showed that iPS\ML/IFN\ associated with MitoTam iodide, hydriodide SKOV3 cells From these data, we hypothesized that inoculation of iPS\ML into the peritoneal cavity of.