Peritoneal lavage with distilled water has been performed during colorectal cancer surgery. This study investigated the cytocidal effects of hypotonic shock in vitro and in vivo in colorectal cancer cells.
Three human colorectal cancer cell lines, DLD1, HT29, and CACO2, were exposed to distilled water, and morphological changes were observed under a differential interference contrast microscope connected to a high-speed digital video camera. Cell volume changes were assessed using a high-resolution flow cytometer. Re-incubation experiments were performed to investigate the cytocidal effects of distilled water. In the in vivo experiment, cancer cells after hypotonic shock were injected intraperitoneally into mice and the degree of established peritoneal metastasis was subsequently evaluated. The effects of the blockade of Cl− channels on these cells during hypotonic shock were also analyzed.
Morphological observations revealed a rapid cell swelling followed by cell rupture. Measurements of cell volume changes showed that mild hypotonic shock induced regulatory volume decrease (RVD) while severe hypotonic shock broke cells into fragments. Re-incubation experiments demonstrated the cytocidal effects of hypotonicity. In vivo experiments revealed the absence of peritoneal dissemination in mice in the distilled water group, and its presence in all mice in the control group. The blockade of Cl− channels increased cell volume by inhibiting RVD and enhanced cytocidal effects during mild hypotonic shock.
These results clearly support the efficacy of peritoneal lavage with distilled water during colorectal cancer surgery and suggest that regulating of Cl− transport may enhance the cytocidal effects of hypotonic shock.
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This work was supported by Grants-in-Aid for Young Scientists (B) (22791295, 23791557), a Grant-in-Aid for Scientific Research (C) (22591464) from the Japan Society for the Promotion of Science, and Research Grant Awards from Kyoto Preventive Medical Center to Dr. Atsushi Shiozaki.
Conflict of interest
The authors declare that no conflict of interest.
K. Takemoto and A. Shiozaki contributed equally to this work.
Electronic supplementary material
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Changes in individual CRC cells subjected to hypotonic stress with distilled water. Quarter speed dynamic images of DLD1 cells subjected to hypotonic stress with distilled water are shown. The cells continued to swell before cell rupture after being subjected to hypotonic stress with distilled water. Similar changes were also observed in HT29 and CACO2 cells (data not shown) (MPEG 6578 kb)
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Takemoto, K., Shiozaki, A., Ichikawa, D. et al. Evaluation of the efficacy of peritoneal lavage with distilled water in colorectal cancer surgery: in vitro and in vivo study. J Gastroenterol 50, 287–297 (2015) doi:10.1007/s00535-014-0971-x
- Colorectal cancer
- Distilled water
- Peritoneal lavage
- Regulatory volume decrease
- Chloride ion transport