Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Phenotype of cancer cells isolated from human CRC tissue. The cytometric analysis of human CRC cells treated with thyroid T3 hormone (0.5 or 1 nM). Samples of patients were divided into groups according to the proportion of CD133 + cells (with Low-er or High-er than median value of general population) and in relation to cancer staging (I–II vs. III–IV). The frequency of cells of given phenotype (A. CD133 + or B. CD133 + CD44 + CD29 + or C. CD133 + CD44 − CD29 + ) is presented on Y-axis (%) and compared with untreated control cells. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Bars and whiskers represent the median with min-max values. *P<0.05 vs. untreated control cells (C), n=6-12 for each experimental group. CRC, colorectal cancer.

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Isolation, MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Phenotype of colorectal cancer cell lines expended in a form of colonospheres. The cytometric analysis of colonospheres derived from (A and C) HCT116 and (B and D) HT29 cell lines and treated with thyroid T3 hormone (0.5 or 1 nM). The frequency of cells of given phenotype is presented on Y-axis (%) and compared with untreated control cells. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Bars and whiskers represent (A and B) the median with min-max values or (C and D) median ± interquartile range *P<0.05 vs. untreated control cells (C), n=6-12 for each experimental group.

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Derivative Assay, MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Sizes of colonospheres. Colonospheres were derived from (A) HCT116 and (B) HT29 cell lines, and (C) cancer cells from human CRC tissues. All were treated with thyroid T3 hormone (0.5, 1 or 2 nM) for 3 days. Samples of patients were divided into two groups according to clinical stage of CRC (I–II vs. III–IV). The diameter of at least 50 spheres of each experimental group was measured with an inverted microscope and a digital camera. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Bars and whiskers represent the median with min-max values. *P<0.05 vs. untreated control cells (C).

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Derivative Assay, Inverted Microscopy, MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Secondary sphere formation ability. Following the T3 treatment spheres derived from (A) HCT116 and (B) HT29 cells or (C) cells isolated from CRC patients were pooled, suspended and seeded in fresh SCM. Spheres were monitored by measuring the maximal outgrowth of sphere's diameter after 1 week. The diameter of at least 20 spheres of each experimental group was measured with an inverted microscope and a digital camera. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Bars and whiskers represent the median with min-max values. *P<0.05 vs. untreated control cells (C).

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Derivative Assay, Isolation, Inverted Microscopy, MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Analysis of cell viability. Colonospheres were derived from (A) HCT116 and (B) HT29 CRC lines and (C) cancer cells isolated from human CRC tissue. They were all treated with thyroid T3 hormone (0.5 and 1 nM). Next, cells were incubated with 7-AAD and analyzed with flow cytometry. Y-axis presents 7-AAD + cells frequency (%) compared with untreated control cells. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Data presented as bars and whiskers representing the median with min-max values. *P<0.05 vs. untreated control cells (C), n=6 for each experimental group.

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Derivative Assay, Isolation, Incubation, Flow Cytometry, MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Distribution of cells in cell cycle phases. Cells of colonospheres formed from (A and D) HCT116 and (B and E) HT29 cell lines and (C and F) cancer cells isolated from human CRC tissue treated with thyroid T3 hormone (0.5 or 1 nM) or untreated control cells (C) were analyzed. (A-C) Y-axis presents cell frequency (%) in active (S/G2/M) or in G0/G1 phases or (D-F) dying cells in subG1 phase. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Data presented as bars and whiskers represent the median with min-max values. *P<0.05 vs. untreated control cells (C), n=6 for each experimental group.

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Isolation, MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Evaluation of (A) viability and (B) proliferation abilities of colorectal cancer cells. Colonospheres formed from HCT116 and HT29 cell lines were treated with thyroid T3 hormone (0.5, 1 and 2 nM) and analyzed. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Bars and whiskers represent median ± interquartile range. *P<0.05 vs. untreated control samples (C), n=6 for each experimental group.

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Western blot analysis of (A) THRα1, (B) THRβ1, (C) DIO2 and (D) DIO3. Densitometric analysis was conducted with HCT116 and HT29 cell lines, and cancer cells isolated from human CRC tissue treated with thyroid T3 hormone (0.5 nM) or untreated control cells (C). Additionally, samples of patients were divided into two groups according to clinical stage of CRC (I–II vs. III–IV). Bars and whiskers represent the median ± interquartile range. Y-axis represents relative protein levels determined by densitometric scanning of the bands and corrected for GAPDH loading control. Statistical significance was showed with Kruskal-Wallis test or Mann-Whitney U test. Each experiment was performed from 3–6 times. *P<0.05. THR, thyroid hormone receptor; DIO, deiodinase.

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Western Blot, Isolation, MANN-WHITNEY

Journal: Oncology Reports

Article Title: Triiodothyronine lowers the potential of colorectal cancer stem cells in vitro

doi: 10.3892/or.2022.8458

Figure Lengend Snippet: Representative western blot analysis images. Western blot analysis of THRα1, THRβ1, DIO2 and DIO3 proteins in HCT116 and HT29 cell lines and cancer cells isolated from human CRC tissue treated with thyroid T3 hormone (0.5 or 1 nM) and compared with untreated control cells (C). Presented sets of bands were obtained in the same western blot membrane. THR, thyroid hormone receptor; DIO, deiodinase.

Article Snippet: Two human CRC cell lines (HT29 and HCT116; (obtained from American Type culture Collection) were used.

Techniques: Western Blot, Isolation

Journal: PLoS ONE

Article Title: Identification of uPAR-positive Chemoresistant Cells in Small Cell Lung Cancer

doi: 10.1371/journal.pone.0000243

Figure Lengend Snippet: Cytotoxic effect of 5-FU on non-sorted and sorted (uPAR-positive and uPAR-negative populations) derived from SCLC cell lines. (A) 1×10 4 cells (H211, H69AR, H1417) were placed in wells of a 48-well plate in triplicates and incubated for 72 hr in the presence of varying concentrations of 5-FU. (B) SCLC cell lines were FACS sorted after staining with anti-uPAR antibodies and were plated at the same seeding density (4×10 3 /well of 96-well plate) and treated with 5-FU at 0, 10, 100, 200 µg/ml for 72 hr. Cell survival was evaluated after adding Guava ViaCount reagent and counting viable and dead cells. Only viable cells were included in data analysis, and 100% viability was defined as number of viable cells cultured in absence of 5-FU. Statistical analysis (2-way ANOVA) of uPAR(+) and uPAR(−) data sets revealed significant differences among viability of uPAR(+) and uPAR(−) cells ( P = 0.0002, 0.0027, 0.0008 for H211, H69AR, H1417 cells, respectively). The data points represent averages±SD of three independent experiments.

Article Snippet: Primary (lung) small cell lung carcinoma (SCLC) cell lines (H1688, H1417, H69AR), bone marrow (BM) metastatic SCLC (H211, H1882) and brain metastatic SCLC (H250) cell lines were obtained from human primary lung and metastatic tissues ( ATCC), grown in RPMI 1640 modified medium (ATCC, N: 30–2001) supplemented with 10% Fetal Bovine Serum (FBS).

Techniques: Derivative Assay, Incubation, Staining, Cell Culture

Journal: PLoS ONE

Article Title: Identification of uPAR-positive Chemoresistant Cells in Small Cell Lung Cancer

doi: 10.1371/journal.pone.0000243

Figure Lengend Snippet: Colony-forming activity of uPAR-positive and uPAR-negative cells derived from SCLC cell lines. (A) H1417- derived, uPAR-positive sorted cells formed multiple colonies in methylcellulose media, while uPAR-negative cells from the same sorts displayed little or no clonogenic activity. (B) Graphical representation of colony-forming ability of uPAR-positive and uPAR-negative cells at different plating densities 3000, 1000, 100 cells/6-well plate (H1417, H69AR, H211). (C) Distribution of uPAR-positive cells in the colonies derived from sorted uPAR-positive cells grown in methylcellulose media. A total of 20 cell colonies from the H1417 cell line were analyzed.

Article Snippet: Primary (lung) small cell lung carcinoma (SCLC) cell lines (H1688, H1417, H69AR), bone marrow (BM) metastatic SCLC (H211, H1882) and brain metastatic SCLC (H250) cell lines were obtained from human primary lung and metastatic tissues ( ATCC), grown in RPMI 1640 modified medium (ATCC, N: 30–2001) supplemented with 10% Fetal Bovine Serum (FBS).

Techniques: Activity Assay, Derivative Assay

Journal: PLoS ONE

Article Title: Identification of uPAR-positive Chemoresistant Cells in Small Cell Lung Cancer

doi: 10.1371/journal.pone.0000243

Figure Lengend Snippet: Expression of CD44 and MDR1 on uPAR-positive and uPAR-negative cells. (A) FACS analysis of, H211, H69AR and H1417 SCLC cell lines double-labeled with uPAR-FITC and CD44-PE, MDR1-PE. The percentages of cells expressing CD44 and MDR1 were calculated separately for uPAR-positive and uPAR-negative cells. (B) Fluorescent microscopic analysis of double-labeled and FACS-sorted cells. Examples of uPAR-FITC/CD44-PE double-labeling (a,b,c) and uPAR-FITC/MDR1-PE double-labeling (d,e,f). (Bf-inset) H1417 cell line stained with mouse IgG isotype control-PE (red), isotype control-FITC (green) and DAPI (blue).

Article Snippet: Primary (lung) small cell lung carcinoma (SCLC) cell lines (H1688, H1417, H69AR), bone marrow (BM) metastatic SCLC (H211, H1882) and brain metastatic SCLC (H250) cell lines were obtained from human primary lung and metastatic tissues ( ATCC), grown in RPMI 1640 modified medium (ATCC, N: 30–2001) supplemented with 10% Fetal Bovine Serum (FBS).

Techniques: Expressing, Labeling, Staining

Journal: BMC Cancer

Article Title: Expression of G-protein inwardly rectifying potassium channels (GIRKs) in lung cancer cell lines

doi: 10.1186/1471-2407-5-104

Figure Lengend Snippet: Expression of GIRK1 in some SCLC cell lines but not in normal SAE cells, nor in adenocarcinoma Clara cell phenotype cell lines . Top panel: GIRK1 is expressed in WBA, H69, and H146 SCLC cell lines but not in H187, H209 and H526 SCLC cell lines. GIRK1 is also not expressed in normal SAE cells, nor is it expressed in H322 or H441 adenocarcinoma Clara cell lines. Bottom panel: Cyclophilin, used as a positive reaction control was seen in all samples. For all gene expression experiments, negative control reactions were performed and found to be negative. The bands on the agarose gels were consistent with the expected sizes: GIRK1-441 bp; cyclophilin-216 bp; M-100 bp.

Article Snippet: The human SCLC cell lines NCI-H69 (H69), NCI-H146 (H146), NCI-H187 (H187), NCI-H209 (H209), and NCI-H526 (H526), the human adenocarcinoma cell lines NCI-H322 (H322), NCI-H441 (H441), and A549, the carcinoid cell line NCI-H727 (H727), and the squamous cell lines NCI-H226 (H226), NCI-H2170 (H2170), and NCI-H520 (H520) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Negative Control

Journal: BMC Cancer

Article Title: Expression of G-protein inwardly rectifying potassium channels (GIRKs) in lung cancer cell lines

doi: 10.1186/1471-2407-5-104

Figure Lengend Snippet: Expression of GIRK3 in SCLC cell lines . Top panel: GIRK3 expression was seen in all six SCLC cell lines, in WBA, H69, H146, H187, H209, and H526. Bottom panel: Cyclophilin, used as a positive reaction control was seen in all samples. For all gene expression experiments, negative control reactions were performed and found to be negative. The bands on the agarose gels were consistent with the expected sizes: GIRK3-317 bp; cyclophilin-216 bp; M-100 bp.

Article Snippet: The human SCLC cell lines NCI-H69 (H69), NCI-H146 (H146), NCI-H187 (H187), NCI-H209 (H209), and NCI-H526 (H526), the human adenocarcinoma cell lines NCI-H322 (H322), NCI-H441 (H441), and A549, the carcinoid cell line NCI-H727 (H727), and the squamous cell lines NCI-H226 (H226), NCI-H2170 (H2170), and NCI-H520 (H520) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Negative Control

Journal: BMC Cancer

Article Title: Expression of G-protein inwardly rectifying potassium channels (GIRKs) in lung cancer cell lines

doi: 10.1186/1471-2407-5-104

Figure Lengend Snippet: Summary of GIRK mRNA expression in lung cells. This is a compilation of data shown in Figures 1–6 & Table 1. Some of the indicated data was not shown.

Article Snippet: The human SCLC cell lines NCI-H69 (H69), NCI-H146 (H146), NCI-H187 (H187), NCI-H209 (H209), and NCI-H526 (H526), the human adenocarcinoma cell lines NCI-H322 (H322), NCI-H441 (H441), and A549, the carcinoid cell line NCI-H727 (H727), and the squamous cell lines NCI-H226 (H226), NCI-H2170 (H2170), and NCI-H520 (H520) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Expression of G-protein inwardly rectifying potassium channels (GIRKs) in lung cancer cell lines

doi: 10.1186/1471-2407-5-104

Figure Lengend Snippet: GIRK1 protein expression in SCLC cells that express GIRK1 mRNA as assessed by western blot analysis . In the three SCLC cell lines that expressed GIRK1 mRNA (Figure 1), membrane protein expression was determined. Top panel: All three SCLC cell lines (WBA, H69, H146) expressed GIRK1 membrane protein. Bottom panel: Actin was used as a control for equal sample loading. The bands are consistent with the expected size: GIRK1-62 kDa; actin-42 kDa.

Article Snippet: The human SCLC cell lines NCI-H69 (H69), NCI-H146 (H146), NCI-H187 (H187), NCI-H209 (H209), and NCI-H526 (H526), the human adenocarcinoma cell lines NCI-H322 (H322), NCI-H441 (H441), and A549, the carcinoid cell line NCI-H727 (H727), and the squamous cell lines NCI-H226 (H226), NCI-H2170 (H2170), and NCI-H520 (H520) were purchased from the American Type Culture Collection (Manassas, VA).

Techniques: Expressing, Western Blot