Detection of mycoplasma infection in circulating tumor cells in patients with hepatocellular carcinoma |
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Authors: | Hong Seo Choi Hyun Min Lee Won-Tae Kim Min Kyu Kim Hee Jin Chang Hye Ran Lee Jae-Won Joh Dae Shick Kim Chun Jeih Ryu |
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Affiliation: | 1. Department of Bioscience and Biotechnology, Institute of Bioscience, Sejong University, Seoul, Republic of Korea;2. Department of Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea;3. Department of Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea;4. Center for Colorectal Cancer, Research Institute and Hospital of National Cancer Center, Goyang-si, Republic of Korea;5. Department of Internal Medicine, Inje University Ilsan Paik Hospital, Goyang-si, Republic of Korea |
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Abstract: | Many studies have shown that persistent infections of bacteria promote carcinogenesis and metastasis. Infectious agents and their products can modulate cancer progression through the induction of host inflammatory and immune responses. The presence of circulating tumor cells (CTCs) is considered as an important indicator in the metastatic cascade. We unintentionally produced a monoclonal antibody (MAb) CA27 against the mycoplasmal p37 protein in mycoplasma-infected cancer cells during the searching process of novel surface markers of CTCs. Mycoplasma-infected cells were enriched by CA27-conjugated magnetic beads in the peripheral blood mononuclear cells in patients with hepatocellular carcinoma (HCC) and analyzed by confocal microscopy with anti-CD45 and CA27 antibodies. CD45-negative and CA27-positive cells were readily detected in three out of seven patients (range 12–30/8.5 ml blood), indicating that they are mycoplasma-infected circulating epithelial cells. CA27-positive cells had larger size than CD45-positive hematological lineage cells, high nuclear to cytoplasmic ratios and irregular nuclear morphology, which identified them as CTCs. The results show for the first time the existence of mycoplasma-infected CTCs in patients with HCC and suggest a possible correlation between mycoplasma infection and the development of cancer metastasis. |
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Keywords: | CTC, circulating tumor cell MAb, monoclonal antibody HCC, hepatocellular carcinoma EMT, epithelial to mesenchymal transition PBMC, peripheral blood mononuclear cell DAPI, 4,6-diamidino-2-phenylindole FBS, fetal bovine serum FITC, fluorescein isothiocyanate PI, propidium iodide RT, room temperature HRP, horse radish peroxidase |
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