Biological characteristics of induction-produced polyploid tumor giant cells in ovarian cancer cell line SKOV3
10.3760/cma.j.cn115355-20231017-00149
- VernacularTitle:卵巢癌细胞株SKOV3中诱导产生的多倍体瘤巨细胞生物学特性
- Author:
Aiqi QIAO
1
;
Xiaoyan YAN
;
Gang LIANG
;
Yanfeng XI
;
Lingmin LI
Author Information
1. 山西医科大学第一医院病理科，太原 030001
- Keywords:
Ovarian neoplasms;
Polyploidy;
Neoplastic stem cells;
Cell differentiation;
Drug resistance
- From:
Cancer Research and Clinic
2024;36(3):199-204
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the morphological and biological characteristics of polyploid tumor giant cells (PGCC) produced by ovarian cancer cell line SKOV3 induced by CoCl 2. Methods:Human ovarian cancer cell line SKOV3 was induced-cultured with 300 μmol/L CoCl 2 in the simulated hypoxic environment for 36 h, the live cells continued to be conventionally cultured and passaged, and the cells collected 20 days later were PGCC group; SKOV3 cell line cultured conventionally was the control group. The formation process and morphological characteristics of PGCC were observed by inverted microscope. The expression of tumor stem cell markers OCT4 and CD117 were detected by immunocytochemistry. The adipogenic differentiation and osteogenic differentiation potential of PGCC were detected by using human bone marrow mesenchymal stem cell adipogenic differentiation assay kit and human bone marrow mesenchymal stem cell osteogenic differentiation assay kit.The cell migration ability of PGCC was detected by scratch assay. PGCC group and control group SKOV3 cells were treated with 1 μmol/L paclitaxel, and the cell morphology of the two groups was observed by microscope at 0, 24 and 48 h to detect the resistance of PGCC to chemotherapy drugs. Results:A small amount of PGCC was observed in SKOV3 cell line cultured in conventional medium under the microscope. CoCl 2 can induce SKOV3 cells to form PGCC, which was nearly round in shape and lacked branching. Its volume was 3 times or more than that of SKOV3 cells, and the nuclei were usually megakaryons or multinucleates, PGCC can produce daughter cells by budding. Immunocytochemical staining showed that OCT4 was positive in some PGCC, but no CD117 was positive. Neither OCT4 nor CD117 was expressed in SKOV3 cells. When cultured with lipid-induced differentiation medium of human bone marrow mesenchymal stem cells, the formation of large vacuoles in the cytoplasm of PGCC was observed at the 3rd cycle, and orange-red, round-like lipid droplets were shown by oil red O staining. Human bone marrow mesenchymal stem cells were cultured in osteogenic induction culture medium for 20 days, and alizarin red staining showed that calcium nodules formed significantly in cells of PGCC group compared with the control group. The cell scratch assay results showed that the migration rates of PGCC cultured in serum-free medium [(59±1)%, (66±3)%] were higher than those of the control group [(11±3)%, (14±5)%] at 24 and 48 h after scratch ( t values were 32.20 and 19.55, both P < 0.001). The migration rates of PGCC cultured in 10% serum medium [(92±3)%, (100±0)%] were higher than those of the control group [(20±6)%, (59±9)%] ( t values were 16.19 and 8.00, both P < 0.001). After 1 μmol/L paclitaxel treatment for 48 h, most of the cells in the PGCC group still survived, while most of the SKOV3 cells in the control group died. Conclusions:PGCC produces daughter cells by budding. PGCC has the characteristics of tumor stem cells: it expresses tumor stem cell markers and has the potential for multidirectional differentiation and strong resistance to chemotherapy drugs.
