Alharbi SA, Ovchinnikov DA, Wolvetang E. Leucine-rich repeat-containing G protein-coupled receptor 5 marks different cancer stem cell compartments in human Caco-2 and LoVo colon cancer lines. World J Gastroenterol 2021; 27(15): 1578-1594 [PMID: 33958845 DOI: 10.3748/wjg.v27.i15.1578]
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Great design and methods used to achieve the aim of the study.
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Thank you very much for your comments.
Reader's ID:
04213496
Submitted on:
April 16, 2021, 11:18
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Reader Comments:
I recognize that LGR5 and BMI1 are representative intestinal stem cell (ISC) markers. At the same time, LGR5 and BMI1 are also known to colorectal cancer stem cell (CSC) markers. This report referred to the cancer stem cells, using famous two cell lines: Caco-2 and Lovo. Therefore, the authors should discuss the relation between colorectal CSCs and LGR5 more in detail in Introduction and discussion. Moreover, the authors did not refer to the relation between colorectal CSCs and BMI1 at all. In contrast, the authors should reduce amount of the description about the relation between ISCs and Lgr5. ISCs did not relate with the main stream of the report.
In discussion, the authors should discuss about underling mechanisms why LGR5-negative cells of Lovo could have stemness nature. For examples, RNA-seq from EYFP-positive or EYFP-negative cells of Lovo could help the understanding of different gene expression profile of CSCs and non-CSCs.
Regarding Cell culture, the authors should confirm the possibility of mycoplasma infection.
Regarding Figure 1I and 1J, I was concerned about stemness properties of LGR5-negative (EYFP-negative) tumor cells. In Figure 1I and 1J, the number of LGR5-negative cells increased day by day. The finding mean that LGR5-negative cells also had a capacity of producing their descendant (LGR5-negative) cells. Therefore, the authors should compare the productivity of daughter cells of LGR5-positive and LGR5-negtive cells (e.g. MTT assay).
Regarding Figure 2, the authors confused ISC markers and CSC markers. The authors should refer to OLFM4, TERT…, as CSC markers, not as ISC markers. And they should displaye the number of samples.
Regarding Figure4, the number of samples was too small (n = 3). The authors should perform xenograft experiments enough times to make a statistics analysis.
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