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Isolated and characterized colorectal cancer stem cells and normal stem cells isolated from colorectal cancer tissues and normal colon tissues by whole genome expression microarray

ABSTRACT: Cancer stem cells (CSCs) are pluripotent cells that have been implicated in tumor proliferation, metastatic dissemination, and recurrence of various cancers. The prognosis of colorectal cancer, which often presents late due to absence of early symptoms, depends on the stage at presentation. In the present study we have isolated and characterized cancer stem cells from colorectal cancer tissues and normal stem cells from normal colon tissues. Only sporadic colorectal cancer cases were considered where the patient had not received any neo-adjuvant therapy to avoid any bias introduced in the cellular transcriptome by external agents. Normal colon tissues were taken from adjacent region of resected specimen as well as non-cancerous operative colon cases. For our study, we have used cancer and normal stem cell markers defined in established literature. For isolating cancer stem cells by FACS, CD44 and CD166 tagged to phycoerythrin (PE) and Alexa Fluor 647 respectively, were used. For isolating normal colon stem cells, CD29, tagged to PE, and Lgr5, tagged to Alexa Fluor 647 were used. Only dual marker-positive cells were isolated to get an enriched population. Tumor sphere assay was done on representative samples to validate the stemness of isolated cells. RNA was isolated from the isolated cell populations and microarray was done to study the whole genome transcriptomic changes.

ORGANISM(S): Homo sapiens

PROVIDER: GSE160988 | GEO | 2023/11/01

REPOSITORIES: GEO

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Project description:Abstract of associated manuscript: The Bacillus subtilis extracytoplasmic function (ECF) sigma(M) factor is activated by cell envelope stress elicited by antibiotics, and by acid, heat, ethanol and superoxide stresses. Here, we have used several complementary approaches to identify genes controlled by sigma(M). In many cases, expression is only partially dependent on sigma(M) because of both overlapping promoter recognition with other ECF sigma factors and the presence of additional promoter elements. Genes regulated by sigma(M) have a characteristic pattern of induction in response to cell envelope-acting antibiotics as evidenced by hierarchical clustering analysis. sigma(M) also contributes to the expression of the Spx transcription factor and thereby indirectly regulates genes of the Spx regulon. Cell envelope stress responses also include regulons controlled by sigma(W), sigma(B) and several two-component regulatory systems (e.g. LiaRS, YycFG, BceRS). Activation of the sigma(M) regulon increases expression of proteins functioning in transcriptional control, cell wall synthesis and shape determination, cell division, DNA damage monitoring, recombinational repair and detoxification. WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van). Each experiment was conducted at least twice using two independent total RNA preparations. For vancomycin untreated and treated experiments, untreated samples were labeled with Alexa Fluor 555 and treated samples with Alexa Fluor 647. For WT vs. mutant experiments, wild type was labeled with Alexa Fluor 555 and mutants with Alexa Fluor 647. For dye swap experiment, wild-type was labeled with Alexa Fluor 647 and mutant with Alexa Fluor 555. Bacillus subtilis CU1065, WT (-van) vs. WT (+van), sigM (-van) vs. sigM (+van), WT (-van) vs. sigM (-van), WT (+van) vs. sigM (+van), WT (-van) vs. spx (-van), WT (+van) vs. spx (+van)

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Isolated and characterized colorectal cancer stem cells and normal stem cells isolated from colorectal cancer tissues and normal colon tissues by whole genome expression microarray

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