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Abstract
The availability of adequate cancer stem cells or cancer stem-like cell (CSC) is important in cancer study. From ovarian cancer cell lines, SKOV3 and OVCAR3, we induced peritoneal ascites tumors in immunodeficient mice. Among the cells (SKOV3.PX1 and OVCAR3.PX1) from those tumors, we sorted both CD44 and CD133 positive cells (SKOV3.PX1_133+44+, OVCAR3.PX1_133+44+), which manifest the characteristics of self-renewal, multi-lineage differentiation, chemoresistance and tumorigenicity, those of cancer stem-like cells (CSLC). Intraperitoneal transplantation of these CD44 and CD133 positive cells resulted in poorer survival in the engrafted animals. Clinically, increased CD133 expression was found in moderately and poorly differentiated (grade II and III) ovarian serous cystadenocarcinomas. The ascites tumor cells from human ovarian cancers demonstrated more CD133 and CD44 expressions than those from primary ovarian or metastatic tumors and confer tumorigenicity in immunodeficient mice. Compared to their parental cells, the SKOV3.PX1_133+44+ and OVCAR3.PX1_133+44+ cells uniquely expressed 5 CD markers (CD97, CD104, CD107a, CD121a, and CD125). Among these markers, CD97, CD104, CD107a, and CD121a are significantly more expressed in the CD133+ and CD44+ double positive cells of human ovarian ascites tumor cells (Ascites_133+44+) than those from primary ovarian or metastatic tumors. The cancer stem-like cells were enriched from 3% to more than 70% after this manipulation. This intraperitoneal enrichment of cancer stem-like cells, from ovarian cancer cell lines or primary ovarian tumor, potentially provides an adequate amount of ovarian cancer stem-like cells for the ovarian cancer study and possibly benefits cancer therapy.
Highlights
Cancer stem cells (CSCs) possess features that promote malignant potential, including cell differentiation, self-renewal, tumorigenicity, chemoresistance and metastasis [1]
PX1 and OVCAR3.PX1 cells based on the expression of CD44 and CD133 and cultured the four types of cells (PX1, PX1_44+, PX1_133+ and PX1_133+44+ cells) in CSC culture media
The SKOV3.PX1_133+44+ and OVCAR3.PX1_133+44+ cells were most capable of rapid proliferation and sphere formation when grown in these ultra-low attachment dishes
Summary
Cancer stem cells (CSCs) possess features that promote malignant potential, including cell differentiation, self-renewal, tumorigenicity, chemoresistance and metastasis [1]. To identify breast CSCs, a subpopulation of primary cancer cells expressing markers of self-renewal and differentiation potential are isolated and tested for tumorigenicity in mouse xenograft models [4]. Bapat et al isolated two clones of ovarian cancer stem-like cells (CSLC) capable of sphere formation from ascites cultures [5], whereas Zhang et al used cell sorting to isolate CD44 and CD117 double-positive ovarian cancer-initiating cells from primary human tumors [6]. Using three human ovarian cancer cell lines and four clinical human primary ascites cell lines, Szotek et al identified a verapamil-sensitive subpopulation that expressed breast cancer resistance protein 1 [7]. An easier method to enrich the cancer stem cells or cancer stem-like cells will facilitate the biological study of cancer stem cells
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