Establishment of a human colorectal cancer cell line P6C with stem cell properties and resistance to chemotherapeutic drugs

Abstract

Aim:Cancer stem cells have the capacity to initiate and sustain tumor growth. In this study, we established a CD44+ colorectal cancer stem cell line with particular emphasis on its self-renewal capacity, enhanced tumor initiation and drug resistance.Methods:Fresh colon cancer and paired normal colon tissues were collected from 13 patients who had not received chemotherapy or radiotherapy prior to surgery. Among the 6 single-cell derived clones, only the P6C cell line was cultured for more than 20 passages in serial culture and formed holoclones with high efficiency, and then the stemness gene expression, colony formation, tumorigenicity and drug sensitivities of the P6C cell line were examined.Results:Stemness proteins, including c-Myc, Oct3/4, Nanog, Lgr5, and SOX2, were highly expressed in the P6C cell line. Oct3/4-positive P6C cells mostly generated holoclones through symmetric division, while a small number of P6C cells generated meroclones through asymmetric division. P6C cells stably expressed CD44 and possessed a high capacity to form tumor spheres. A single cell-derived sphere was capable of generating xenograft tumors in nude mice. Compared to SW480 and HCT116 colorectal cancer cells, P6C cells were highly resistant to Camptothecin and 5-fluorouracil, the commonly used chemotherapeutic agents to treat colorectal cancers.Conclusion:We established a colorectal cancer stem cell line P6C with a high tumorigenic capacity and the characteristics of normal stem cells. It will benefit the mechanistic studies on cancer stem cells and the development of drugs that specifically target the cancer stem cells.

Role of iron oxide core of polymeric nanoparticles in the thermosensitivity of colon cancer cell line HT-29

“…On day 10, spheroids with mean diameter of 100 mm were treated with 1, 5 or 10 mM of 5-FU (Sigma) [22] or the same concentration of 5-FU released from 5-FU-loaded PLGAcoated nanoparticles with or without the iron oxide core for 71 h. PLGA nanoparticles were used as a control for the drugloaded nanoparticles treatment. Before heat treatment, the medium was replaced with fresh RPMI 1640 culture medium.…”

Reduced CD146 expression promotes tumorigenesis and cancer stemness in colorectal cancer through activating Wnt/β-catenin signaling

“…P6C is a CD44 hi primary CRC cell line of tumor-initiating potential (Supplementary Figure S1) [42]. Furthermore, CD44 + CD133 + cell population has been reported to enrich in colorectal tumor-initiating cells [43, 44].…”

Cancer stem cells in human digestive tract malignancies

“…Besides introduced markers listed in Table 1 Apart from in vitro studies, analyzing clinical samples revealed that CD44 could be used in combination with clinicopathological information to improve the OSCC prognosis, since its overexpression, in tumor samples and peripheral blood of OSCC patients, was correlated to poor prognosis LGR5 [152,153] EpCAM [120] BMI1 [119,218] CD24 [122,125] OCT4 [118,119] SOX2 [118,119,[149][150][151] NANOG [118,119] Colon and rectum CD44 [166,[180][181][182][183][184][185][186][187][188][189][190][191][192][193][194][195][196][197][198]200] CD133 [161-165, 171-178, 186-188, 232] ABCG2 [75,158,159] ALDH1 [198,[222][223][224]…”

“…LGR5 has also been considered as a CRC-SC marker, because spheroid cultures derived from primary tumors were enriched for LGR5 expression [199,200], and LGR5 + cells form CRC cell lines displayed colony forming, tumorigenic [201], and therapy resistance [202] abilities. These reports are in agreement with other experiments indicating a significant decrease in survival [203], drug resistance [204], proliferation, migration, and tumorigenic abilities [205] of CRC cells upon LGR5 knockdown.…”