sult1a1 cdna  (OriGene)

Journal: medRxiv

Article Title: FIS103, a Novel SULT1A1-dependent Prodrug, Demonstrates Potent Antitumor Activity in Renal Cell Carcinoma

doi: 10.1101/2024.03.21.24304257

Figure Lengend Snippet:

Article Snippet: The membrane was incubated with SULT1A1 primary antibody (R&D Systems, Cat no: MAB5546) diluted in blocking buffer overnight at 4°C.

Techniques: Expressing

Journal: medRxiv

Article Title: FIS103, a Novel SULT1A1-dependent Prodrug, Demonstrates Potent Antitumor Activity in Renal Cell Carcinoma

doi: 10.1101/2024.03.21.24304257

Figure Lengend Snippet: A) Kaplan-Meier Plots indicating that high expression of SULT1A1 mRNA correlates with poor prognosis for patients with i) kidney renal clear cell carcinoma, ii) brain lower grade glioma, and iii) uveal melanoma. Percentage of patients with B) high SULT1A1 mRNA expression in RCC subtypes (RNASeq from the TCGA PanCancer study), and C) with SULT1A1 mutation (whole genome sequencing from the TCGA PanCancer study and Genentech 2014 study). All TCGA data was extracted via cBioPortal. The number of cases per RCC subtypes is as followed: clear cell RCC (ccRCC) n=512, papillary RCC (pRCC) n=283, chromophobe RCC (chrRCC) n=65, non-clear cell RCC (nccRCC) n=146.

Article Snippet: The membrane was incubated with SULT1A1 primary antibody (R&D Systems, Cat no: MAB5546) diluted in blocking buffer overnight at 4°C.

Techniques: Expressing, Mutagenesis, Sequencing

Journal: medRxiv

Article Title: FIS103, a Novel SULT1A1-dependent Prodrug, Demonstrates Potent Antitumor Activity in Renal Cell Carcinoma

doi: 10.1101/2024.03.21.24304257

Figure Lengend Snippet: A) SULT1A1 mRNA levels in SULT1A1 positive (T47D) and SULT1A1 negative (MDA-MB-231) cell lines. B) FIS03 demonstrated differential cell killing (cell viability assay) in T47D but not MDA-MB-231 cells, showing SULT1A1 dependency. C) The differential cytotoxic effect of FIS103 was dose dependent (concentration is nM). DMSO was used as vehicle control.

Article Snippet: The membrane was incubated with SULT1A1 primary antibody (R&D Systems, Cat no: MAB5546) diluted in blocking buffer overnight at 4°C.

Techniques: Viability Assay, Concentration Assay, Control

Journal: medRxiv

Article Title: FIS103, a Novel SULT1A1-dependent Prodrug, Demonstrates Potent Antitumor Activity in Renal Cell Carcinoma

doi: 10.1101/2024.03.21.24304257

Figure Lengend Snippet: A) Proposed mechanism of action of N-BIC compounds after activation by SULT1A1 enzyme. B) Aligned and superimposed representative structures of SULT1A1: 1LS6 (green) and 2D06 (grey). C) Cluster representative structure of FIS103 during 100ns MD simulation with i) 2D06 and ii) ILS6.

Article Snippet: The membrane was incubated with SULT1A1 primary antibody (R&D Systems, Cat no: MAB5546) diluted in blocking buffer overnight at 4°C.

Techniques: Activation Assay

Journal: medRxiv

Article Title: FIS103, a Novel SULT1A1-dependent Prodrug, Demonstrates Potent Antitumor Activity in Renal Cell Carcinoma

doi: 10.1101/2024.03.21.24304257

Figure Lengend Snippet: A) SULT1A1 mRNA and protein expression in RCC SULT1A1 high- and SULT1A1 low-expressing cell lines. B) Cell viability assay demonstrating FIS103 treatment in SULT1A1 high-expression RCC cells. C) Cell viability assay demonstrating lack of toxicity of FIS103 up to 1 uM on cells with low SULT1A1 expression. D) Cell viability assay demonstrating lack of toxicity of FIS103 versus the parental compound (NSC-743380) in SULT1A1 non-expressing cells (MDA-MB-231). E) 6-week old NU/J mice were injected with 10 6 A498 cells to allow tumor growth for 20 days before intraperitoneal injection of FIS103 at indicated concentrations for 14 days (once daily). Test groups for mice (table) and relative weights by treatment group (blue, vehicle; orange, 25 mg/kg; grey, 50 mg/kg then 10 mg/kg) over the course of the 41-day study. F) Tumor volume was measured over the course of the study (left). Control group tumors grew for 20 days and were then sacrificed due to excess tumor burden. Treatment groups were started at 24 mg/kg and 50 mg/kg doses. Mice getting 50 mg/kg showed signs of toxicity, and 2 died. The rest of this group were given a 3 day drug holiday and then treated at 10 mg/kg. All mice in the treatment groups had tumors that became non-detectable after 14 days and remained absent through study conclusion. Representative photos of the mice in each group are shown (right).

Article Snippet: The membrane was incubated with SULT1A1 primary antibody (R&D Systems, Cat no: MAB5546) diluted in blocking buffer overnight at 4°C.

Techniques: Expressing, Viability Assay, Injection, Control

Journal: PLoS ONE

Article Title: Metabolic Patterns and Biotransformation Activities of Resveratrol in Human Glioblastoma Cells: Relevance with Therapeutic Efficacies

doi: 10.1371/journal.pone.0027484

Figure Lengend Snippet: (A) ICC evaluation of SULT1A1, 1C2 and 4A1 expression in PBC, UW228-3 and LN-18 cells without (N) and with (R) resveratrol treatment. (B) Western blot analyses of SULT1A1, 1C2 and 4A1 expression in LN-18 cells without (N) and with (R) resveratrol treatment and compared with that in normal control PBCs and resveratrol-sensitive control medulloblastoma UW228-3 cells. β -actin was used as loading control and for calculation of SULT expression levels/densitometry scan of Western blotting results. *Compared with normal cultured LN-18 and UW228-3 cells, respectively; *represents statistical significance ( p <0.05).

Article Snippet: ICC staining and Western blot analysis were performed on the samples obtained from each of the experimental groups using the rabbit anti-human SULT1A1, 1C2 and 4A1 antibodies (ProteinTech Group, Inc., Chicago, USA) by the method described elsewhere – .

Techniques: Expressing, Western Blot, Cell Culture

Journal: PLoS ONE

Article Title: Metabolic Patterns and Biotransformation Activities of Resveratrol in Human Glioblastoma Cells: Relevance with Therapeutic Efficacies

doi: 10.1371/journal.pone.0027484

Figure Lengend Snippet: Immunohistochemical illustration of differential expression patterns of SULT1A1, 1C2 and 4A1 in two cases of glioblastomas (GM). Medulloblastoma (MB) and its surrounding noncancerous cerebellum tissue (NC) were cited as controls.

Article Snippet: ICC staining and Western blot analysis were performed on the samples obtained from each of the experimental groups using the rabbit anti-human SULT1A1, 1C2 and 4A1 antibodies (ProteinTech Group, Inc., Chicago, USA) by the method described elsewhere – .

Techniques: Immunohistochemical staining, Expressing

Journal: PLoS ONE

Article Title: Metabolic Patterns and Biotransformation Activities of Resveratrol in Human Glioblastoma Cells: Relevance with Therapeutic Efficacies

doi: 10.1371/journal.pone.0027484

Figure Lengend Snippet: (A) Immunofluorescence illustration of intracellular distribution of STAT3 in LN-18 and PBC cells without (N) and with (AG490) 60 µM AG490 treatment. Arrows indicate the cells shown in the insets in high magnification (X400). (B) Flow cytometry analysis revealed reduction of G1-phase cells, accumulation of S-phase cells and induction of apoptosis (blue peak) in AG490-treated LN-18 cell population. The cell cycle progression was almost unchanged in PBC cells. *, indicates the peak of apoptotic cells. (C) H&E morphologic examination of LN-18 cells under normal culture or incubated with 100 µM trans -resveratrol, 60 µM AG490 or resveratrol/AG490 mixture for 48 hours (Main images). Small images: immunocytochemical illustration of SULT1A1, 1C2 and 4A1 expression in LN-18 cells treated by 60 µM AG490 without/with 100 µM resveratrol supplementation. Normally cultured and 100 µM resveratrol-treated LN-18 cells were cited as controls.

Article Snippet: ICC staining and Western blot analysis were performed on the samples obtained from each of the experimental groups using the rabbit anti-human SULT1A1, 1C2 and 4A1 antibodies (ProteinTech Group, Inc., Chicago, USA) by the method described elsewhere – .

Techniques: Immunofluorescence, Flow Cytometry, Incubation, Expressing, Cell Culture

Journal: Molecular cancer therapeutics

Article Title: Decreased DNA damage and improved p53 specificity of RITA analogs

doi: 10.1158/1535-7163.MCT-22-0119

Figure Lengend Snippet: (A) Chemical structures of RITA, NSC777196, NSC782846. (B) Correlation of SULT1A1 mRNA level with sensitivity to RITA, NSC777196 and NSC782846 in NCI-60 cell line database. (C) Upper panels, SULT1A1 protein level detected by Western blot in seven cancer cell lines, including breast carcinoma T47D, MDA-MB-231; skin cancer A431; osteosarcoma U2OS, SJSA; melanoma A375; and lung carcinoma H1299. β-Actin was used as loading control. Lower panels, NSC777196 efficiently suppressed the growth of high SULT1A1-expressing cells (U2OS, T47D and A431, red lines) while having negligible effect in cells with low SULT1A1 (A375, MDA-MB-231, SJSA, H1299, black lines). The antitumor effect of NSC782846 was less dependent on the level of SULT1A1 expression. Shown are the results obtained from five biological replicates. (D) Cell viability of WT and SULT1A1−/− MCF7 cells after 24 hours treatment with RITA, NSC777196 and NSC782846 (n=5, * p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: A375 and SJSA cells, stably expressing SULT1A1 cDNA (OriGene, #RC201601L1), were generated by lentivirus transduction as described previously ( 16 ).

Techniques: Western Blot, Expressing

Journal: Molecular cancer therapeutics

Article Title: Decreased DNA damage and improved p53 specificity of RITA analogs

doi: 10.1158/1535-7163.MCT-22-0119

Figure Lengend Snippet: (A) mRNA level of representative oncogenes after 16 hours treatment of MCF7 SULT1A1−/− cells with 10μM NSC777196 or 10μM NSC782846 measured by qPCR (n=3 * p<0.05, ** p<0.01, ***p<0.001). (B) Downregulation of selected oncogenes upon treatment of MCF7 SULT1A1−/− cells by 10μM NSC777196 and 10μM NSC782846 as detected by Western blotting. (C) Protein levels of selected oncogenes in MCF7 SULT1A1−/− in which p53 was depleted by 2 different shRNAs treated with 10μM NSC777196 or 10μM NSC782846 for 24 hours. Scrambled shRNA and GFP shRNA were used as controls. (D) Apoptotic cells were detected by Annexin V-PI double staining using FACS after treatment of MCF7 SULT1A1 −/− cells with 10μM NSC777196 or 10μM NSC782846 for 16 hours. Representative pictures are shown in left panel. The total number of cells in the Q2 and Q3 quadrant was regarded as apoptotic cells. Percentages of apoptotic cells are shown in the bar graph (right panel; n=3 * p<0.05).

Article Snippet: A375 and SJSA cells, stably expressing SULT1A1 cDNA (OriGene, #RC201601L1), were generated by lentivirus transduction as described previously ( 16 ).

Techniques: Western Blot, shRNA, Double Staining

Journal: Molecular cancer therapeutics

Article Title: Decreased DNA damage and improved p53 specificity of RITA analogs

doi: 10.1158/1535-7163.MCT-22-0119

Figure Lengend Snippet: (A) DNA damage marker γH2AX and p53 detected by Western blotting upon 24-hour treatment of MCF7 SULT1A1−/− cells with 1μM RITA, or 10μM and 25μM of NSC777196 or NSC782846, respectively. (B) DNA damage marker γH2AX detected by immunostaining upon treatment by RITA (1μM), NSC777196 (10μM) and NSC782846 (10μM) in MCF7 SULT1A1 −/− cells. (C) RNA synthesis monitored by EU incorporation in MCF7 SULT1A1−/− cells treated with RITA (1μM), NSC777196 (10μM), NSC782846 (10μM), and ActD (2.5 mM), then incubated for 1 hour with 1 mM EU. EU was labeled by click chemistry and visualized by fluorescent microscopy. (D) Protein levels of total RNA Pol II and phospho-Ser2 RNA Pol II in MCF7 SULT1A1−/− cells treated with RITA (1μM), NSC777196 (10μM), NSC782846 (10μM) for 24 hours. (E) The levels of representative oncogenes and RNA Pol II upon treatment with low dose of NSC777196 and NSC782846 in MCF7 WT cells as detected by Western blotting. (F) RNA synthesis monitored by EU incorporation in MCF7 WT cells treated with NSC777196 (0.25μM), NSC782846 (1μM), then incubated for 1 hour with 1 mM EU. EU was labeled by click chemistry and visualized by fluorescent microscopy.

Article Snippet: A375 and SJSA cells, stably expressing SULT1A1 cDNA (OriGene, #RC201601L1), were generated by lentivirus transduction as described previously ( 16 ).

Techniques: Marker, Western Blot, Immunostaining, Incubation, Labeling, Microscopy