Differentiation of a Cell Line of Human Cervical Argyrophil Small Cell Carcinoma to Macrophage Lineage Cells

Abstract

To investigate the origin of argyrophil small cell carcinoma (ASCC) of the uterine cervix, we examined the influence of dibutyryl cyclic adenosine 3′ ′ ′ ′,5′ ′ ′ ′-monophosphate (dB-cAMP), a known differentiation inducer, on the characteristics of an ASCC cell line, TC-YIK, which has been shown to be a useful in vitro experimental model of ASCC. In TC-YIK cells after treatment with dB-cAMP, two specific antigenic markers of macrophages, CD14 and human leukocyte antigen-DR, were detected by flow cytometric analysis. In addition, interferon-γ γ γ γ mRNA was detected by reverse transcription-polymerase chain reaction and interferon-γ γ γ γ protein was detected by ELISA. Argyrophil small cell carcinoma (ASCC) of the uterine cervix, first described by Albores-Saavedra et al., 1) is characterized morphologically by the presence of argyrophil granules in the cytoplasm of the undifferentiated small cells and clinically by rapid metastasis and a poor prognosis. More than1-7) Because ASCC is a rare cervical tumor, accounting for only 1.5 to 5% of all cervical tumors, [1][2][3][4][5][6][7] most reports on ASCC have been concerned with one or a few cases. Therefore, development of in vivo and in vitro experimental systems is desirable not only for examining the biological behavior of ASCC, but also for establishing an effective clinical treatment.For this purpose we previously established and characterized an ASCC tumor line in nude mice and a cell line from the tumor, designated TC-YIK. [8][9][10] In our previous studies, we found that TC-YIK cells retained the histochemical characteristics of ASCC cells and could be used as an in vitro experimental model of ASCC, and that the TC-YIK cells were possibly of neuroendocrine origin, based on electron microscopic evidence of small, electron-dense, membrane-bound neurosecretory-type granules and immunohistochemical evidence of neuron-specific enolase, chromogranin, serotonin and gastrin. 8,9,11) In terms of the origin of ASCC, however, there have been three theories: ASCC originates from (i) amine precursor uptake and decarboxylation (APUD) cells, 2,3,12) (ii) undifferentiated epithelial cells with multi-differentiation ability, 13) or (iii) stem cells. 7)In this study, to investigate the origin of ASCC of the uterine cervix and the feasibility of treatment of the tumor with a differentiation inducer, we examined the influence of dibutyryl cyclic adenosine 3′,5′-monophosphate (dBcAMP), a known differentiation inducer, on the characteristics of an ASCC cell line, TC-YIK. MATERIALS AND METHODSCell culture TC-YIK cells were grown in RPMI-1640 medium (Nikken Biomedical Lab., Kyoto) supplemented with 10% bovine calf serum (FBS; Cam Sera, Ontario, Canada) under conditions of 5% CO 2 and saturated humidity at 37°C. Isolation and characterization of TC-YIK cells are described elsewhere.9, 10) TC-YIK cells were cloned twice after 60 cell generations from the start of culture using the limiting dilution method. The clone that showed the most stable growth was used for the current study. ...

Establishment of the cell line, HeLa-CD14, transfected with the human CD14 gene

“…Although LPS is considered as its main ligand, CD14 also recognizes other pathogen-associated molecular patterns (PAMPs). Studies have shown that dibutyryl cyclic adenosine 3' ,5'-monophosphate (dB-cAMP) promoted the differentiation of argyrophil small cell carcinoma cells (ASCC) of the uterine cervix to macrophages, with two specific antigenic markers, CD14 and human leukocyte antigen-DR (9). M2 macrophages are transformed to activated M1 macrophages by CD4 + Th1 cells under the influence of prostaglandin E2 and IL-6 in cervical carcinoma (10).…”