Advanced colorectal cancer (CRC) is highly metastatic and often results in peritoneal dissemination. The extracellular vesicles (EVs) released by cancer cells in the microenvironment are important mediators of tumor metastasis. We investigated the contribution of EV-mediated interaction between peritoneal mesothelial cells (MCs) and CRC cells in generating a pro-metastatic environment in the peritoneal cavity. Peritoneal MCs isolated from peritoneal lavage fluids displayed high CD44 expression, substantial mesothelial-to-mesenchymal transition (MMT) and released EVs that both directed tumor invasion and caused reprogramming of secretory profiles by increasing TGF-β1 and uPA/uPAR expression and MMP-2/9 activation in tumor cells. Notably, the EVs released by tumor cells induced apoptosis by activating caspase-3, peritoneal MC senescence, and MMT, thereby augmenting the tumor-promoting potential of these cells in the peritoneal cavity. By using pantoprazole, we reduced the biogenesis of EVs and their pro-tumor functions. In conclusion, our findings provided evidence of underlying mechanisms of CRC dissemination driven by the interaction of peritoneal MCs and tumor cells via the EVs released in the peritoneal cavity, which may have important implications for the clinical management of patients.

The Interaction between Reactive Peritoneal Mesothelial Cells and Tumor Cells via Extracellular Vesicles Facilitates Colorectal Cancer Dissemination.

The invention discloses a constructed human primary colorectal cancer liver metastases cell strain HCS1220. In vitro culture of the human primary colorectal cancer liver metastases cell strain HCS1220in a culture medium can be realized; in vitro culture growth is rapid and stable; continuous subculture can be realized; it is convenient for construction of a metastatic colon carcinoma generation mechanism model; the immunodeficient mice subcutaneous tumorigenicity is excellent; growth of tumor with time passing is rapid, and the wettability is high; the excellent tumorigenicity is capable of providing importance guarantee for drug in vivo tests; the human primary colorectal cancer liver metastases cell strain HCS1220 can be taken as a cell material of colorectal cancer liver metastases pathogenesis and individual treatment in vitro study, and providing base for preparation, screening, and evaluation of antitumor drugs.

Human primary colorectal cancer liver metastases cell strain HCS1220

To establish one primary cell line of human liver metastasis of colon cancer. HCS1220 cell line was derived from one liver metastasis of colon cancer patient’s resected tumor sample. The characterization of the cell line was defined by karyotype analysis, short tandem repeat (STR) analysis and mycoplasma contamination. Subcutaneous injection 1 × 106 cells to four BALB/c nude mice, the viable tumors were developed and diagnosed (H&E staining). The expression of biomarkers CK20 and CDX2 for colon cancer were determined by immunocytochemistry assay. HCS1220 cell line can grow stably and continuously passage. During the grow process, the contact loss in the growth process and superimposed growth, which could be defined as proliferation of malignant tumor. Chromosome analysis revealed the cells derived from human female. The cells were not contaminated by mycoplasma. By immunohistochemistry, the cell line was proven to express the biomarkers of colon cancer CK20 and CDX2, while a-fetoprotein, hep-1 and glypican-3 were stained negative, which demonstrated that the HCS1220 cell line originating from the intestinal tissue. HCS1220 cell line has the characteristics of primary human liver metastasis of colon cancer. The results of STR have genetically showed that cell line is original, which can provided cell materials for research in vitro and can also help for establishing the mechanism model of liver metastasis of colon cancer and preparing, screening and evaluating anti-tumor drugs.

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Yi Chai

Papers

Establishment and characterization of a cell line HCS1220 from human liver metastasis of colon cancer