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Combined Conditional Knockdown and Adapted Sphere Formation Assay to Study a Stemness-Associated Gene of Patient-derived Gastric Cancer Stem Cells
Published on: May 9, 2020
Establishment and characterization of GCSR1, a multi-drug resistant signet ring cell gastric cancer cell line
Xin Xu1, Li-Juan Qian2, Xing-Yun Su1
1Department of Surgical Oncology, The 1st Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310003, P.R. China.
Abstract:
Signet ring cell gastric cancer (SRCGC) has very poor prognosis worldwide, and studying its molecular characteristics is urgent for improving the outcome. However, few well-characterized SRCGC cell lines are available for research. Therefore, we established a novel cell line GCSR1, from a Chinese male SRCGC patient. Cell morphology of GCSR1 in culture, maintained in vitro for over 90 passages, is similar to the cells from the patient. GCSR1 cells proliferated in vitro with a doubling time of 67.65 h. Karyotyping showed they were aneuploid. Missense mutation occurred in codon 193 of P53 and deletion occurred in exons 1 and 3 of P16. Results of CCK8 assay revealed that GCSR1 was more resistant to 5-fluorouracil (5-FU) and mitomycin (MMC) than other gastric cancer cell lines. Stem cell marker assay by flow cytometry showed that GCSR1 had high proportion of CD44+ and/or CD133+ cells. It formed colonies easily in soft agar and generated xenograft tumors in nude mice. In conclusion, GCSR1 is a well-established, well-characterized multi-drug resistant cell line with abundant cancer stem cells.
Insights
Researchers developed GCSR1, a new cell line for signet ring cell gastric cancer (SRCGC). This well-characterized model exhibits multi-drug resistance and cancer stem cell properties, aiding SRCGC research.
Area of Science:
- Oncology
- Cell Biology
- Cancer Research
Background:
- Signet ring cell gastric cancer (SRCGC) presents a poor prognosis globally.
- Limited availability of well-characterized SRCGC cell lines hinders research progress.
Purpose of the Study:
- Establish and characterize a novel cell line for SRCGC research.
- Provide a valuable tool for investigating SRCGC molecular mechanisms and treatment resistance.
Main Methods:
- Established a new cell line (GCSR1) from a Chinese male SRCGC patient.
- Performed cell morphology, proliferation, karyotyping, and drug resistance assays (CCK8).
- Assessed stem cell markers (CD44, CD133) via flow cytometry and evaluated tumorigenicity in vivo.
Main Results:
- GCSR1 cells maintained patient-like morphology over 90 passages and exhibited aneuploidy.
- Identified P53 and P16 gene mutations (missense and deletion, respectively).
- Demonstrated significant resistance to 5-fluorouracil (5-FU) and mitomycin (MMC), high CD44+/CD133+ cell populations, and xenograft formation.
Conclusions:
- GCSR1 is a well-established, multi-drug resistant cell line with abundant cancer stem cells.
- This novel cell line serves as a robust model for studying SRCGC.
- GCSR1 facilitates research into SRCGC pathogenesis and the development of targeted therapies.
