Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Primer pairs used to amplify polymerase chain reaction ( PCR ) products

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Polymerase Chain Reaction

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Tissue array analysis of SULT1E1, PAPSS1 and PAPSS2 in 30 cases of breast cancer. Representative staining of SULT1E1 (a,b), PAPSS1 (d,e) and PAPSS2 (g,h) in the adjacent normal breast tissues and tumorous tissues were shown respectively. The immunohistochemical scores of SULT1E1, PAPSS1 and PAPSS2 in the breast tissue assay are summarized as the normalized means of scores ± standard error (SE) in (c,f) and (i), respectively. **P < 0.01 versus adjacent normal breast tissues.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Staining, Immunohistochemical staining

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Tissue array analysis of SULT1E1, PAPSS1 and PAPSS2 in 30 cases of endometrial cancer. Representative staining of SULT1E1 (a,b), PAPSS1 (d,e) and PAPSS2 (g,h) in the adjacent normal endometrial tissues and tumorous tissues were shown, respectively. The immunohistochemical scores of SULT1E1, PAPSS1 and PAPSS2 in the endometrial tissue assay are summarized as the normalized means of scores ± standard error (SE) in (c,f) and (i), respectively. *P < 0.05 versus adjacent normal endometrial tissues; **P < 0.01 versus adjacent normal endometrial tissues.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Staining, Immunohistochemical staining

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Expressions of SULT1E1, PAPSS1 and PAPSS2 enzymes in clinical breast tissue samples. The mRNA and protein expressions in normal, para‐tumor and tumor tissues are shown in (a) and (b), respectively. The protein levels were normalized to the internal control and represented as the means of results from different samples ± standard error (SE) (c). **P < 0.01 versus normal tissues.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Control

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: The effect of SULT1E1 on the tumorigenesis in subcutaneous xenograft model after the injection of adenovirus‐SULT1E1 around the tumor. The mean body weight in tumor of nude mice is shown in (a). The tumor growth curve was represented in (b). *P < 0.05 versus the day of first injection.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Injection

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Effects of SULT1E1 and/or PAPSS1 overexpression on cell growth and apoptosis in MCF‐7 cells in the absence or presence of 17β‐estradiol (E 2). The overexpression of SULT1E1 and PAPSS1 by adenovirus in MCF‐7 cells were confirmed by Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) and western‐blot (a–c). MCF‐7 cells infected with Ad‐GFP, Ad‐SULT1E1, Ad‐PAPSS1 or Ad‐SULT1E1 mixed with Ad‐PAPSS1 were cultured without or with 1, 10, 100 nM, 1 μM E 2. The cell proliferation was assessed by CCK‐8 assay. Panel (d) shows the cell growth of MCF‐7 cells in the absence or presence of 10 nM E 2 after 24 and 48 h culture. Panel (e) presents the growth curve of MCF‐7 cells with SULT1E1 and/or PAPSS1 overexpression in the presence of different concentrations of E 2 for 24 h. 1 mM H 2 O 2 was used to induce apoptosis of MCF‐7 cells for 6 h in the absence or presence of 10 nM E 2. The ratio of apoptotic cells in 1 × 106 total cells was analyzed by flow cytometry after incubation with Annexin V‐APC and 7‐ADD and summarized in (f). The data represents the mean of three independent experiments ± standard deviation (SD). *P < 0.05 versus Ad‐GFP group; #P < 0.05 versus N/A group.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Over Expression, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Infection, Cell Culture, CCK-8 Assay, Flow Cytometry, Incubation, Standard Deviation

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Effects of SULT1E1 and/or PAPSS1 overexpression on the mRNA levels of proliferation and apoptosis related genes in MCF‐7 cells in the absence of 17β‐estradiol. MCF‐7 cells infected with Ad‐GFP, Ad‐SULT1E1, Ad‐PAPSS1 or Ad‐SULT1E1 mixed with Ad‐PAPSS1 were cultured in medium without 17β‐estradiol for 48 h. The mRNA levels of c‐myc, cyclin D1, bcl‐2 and bax were analyzed by Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) and summarized in (a–d). The data represent the mean of three independent experiments ± standard deviation (SD). *P < 0.05 versus N/A group.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Over Expression, Infection, Cell Culture, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Effects of SULT1E1 and/or PAPSS1 overexpression on the mRNA levels of proliferation and apoptosis related genes in MCF‐7 cells in the absence or presence of 17β‐estradiol. After treatment with Ad‐GFP, Ad‐SULT1E1, Ad‐PAPSS1 or Ad‐SULT1E1 mixed with Ad‐PAPSS1 for 48 h, MCF‐7 cells were incubated without or with 1, 10, 100 nM, 1 μM E 2 for 6 h. The mRNA levels of c‐myc, cyclin D1, bcl‐2 and bax were analyzed by Real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) and summarized in (a–d). The data represent the mean of three independent experiments ± standard deviation (SD). *P < 0.05 versus N/A group.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Over Expression, Incubation, Reverse Transcription, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Standard Deviation

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Effects of SULT1E1 and/or PAPSS1 overexpression on the protein levels of proliferation and apoptosis related genes in MCF‐7 cells in the absence or presence of 17β‐estradiol. After treatment with Ad‐GFP, Ad‐SULT1E1, Ad‐PAPSS1 or Ad‐SULT1E1 mixed with Ad‐PAPSS1 for 48 h, MCF‐7 cells were incubated without or with 1, 10, 100 nM, 1 μM E 2 for 18 h. The protein levels of c‐myc (a), cyclin D1 (c), bcl‐2 (e) and bax (g) were analyzed by western‐blot. Summaries of three experiments are shown in (b,d,f,h). The data represent the mean of three independent experiments ± standard deviation (SD). *P < 0.05 versus N/A group.

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Over Expression, Incubation, Western Blot, Standard Deviation

Journal: Cancer Science

Article Title: Effect of estrogen sulfation by SULT 1 E 1 and PAPSS on the development of estrogen‐dependent cancers

doi: 10.1111/j.1349-7006.2012.02258.x

Figure Lengend Snippet: Estrogen sulfation by Ad‐SULT1E1 and/or Ad‐PAPSS1. The formation of sulfated estrogen was analyzed by alkaline–chloroform extraction. MCF‐7 cells were cultured in a 24‐well plate and infected with Ad‐GFP, Ad‐SULT1E1, Ad‐PAPSS1 or Ad‐SULT1E1 mixed with Ad‐PAPSS1 for 48 h. Then the cells were incubated with 10 nM [3 H] E 2 in the absence or presence of 1, 10, 100, 1000 nM cold estrogen for the indicated time. Sulfated estrogen in the medium was measured by liquid scintillation counting as described in Materials and Methods. The E 2 S formation in time‐course is shown in (a). The ratio of E 2 S to E 2 in cultured medium is shown in (b).

Article Snippet: Rabbit anti‐human polyclonal antibody against SULT1E1 (1:25 dilution; Proteintech Group, Chicago, IL, USA), rabbit anti‐human polyclonal antibody against PAPSS2 (1:25 dilution; Abcam, Cambridge, MA, USA), goat anti‐human polyclonal antibody against PAPSS1 (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and rabbit anti‐human polyclonal antibody Estrogen Receptor α (ER α, 1:50 dilution; Cell Signal Technology, Boston, MA, USA) were used as the first antibodies.

Techniques: Extraction, Cell Culture, Infection, Incubation

Journal: bioRxiv

Article Title: Glycosaminoglycan-mediated lipoprotein uptake protects cancer cells from ferroptosis

doi: 10.1101/2024.05.13.593939

Figure Lengend Snippet: a. Number of doublings (log 2 ) in 5 days of the indicated Karpas299 (left) and A-498 (right) UGDH _KO cell lines expressing a sgRNA resistant UGDH cDNA (blue) or an empty vector (grey) under the indicated concentrations of ferrous ammonium citrate (FAC). b. Immunoblot analysis of TFRC (top) and IRP1 (bottom) in the indicated UGDH _KO cell lines expressing a sgRNA resistant UGDH cDNA or an empty vector treated or not with FAC (0.1 mg/mL). GAPDH is included as a loading control. c. Immunoblot analysis of GPX4 (left), SLC7A11 (center), and AIFM2 (right) in the indicated UGDH _KO cell lines expressing a sgRNA resistant UGDH cDNA or an empty vector. GAPDH is included as a loading control. d. Representative flow cytometry plot showing the uptake of DiI-LDL (PE median intensity) in Karpas299 UGDH _KO cells expressing a sgRNA resistant UGDH cDNA (blue) or an empty vector (grey). e. Cellular uptake of DiI-LDL measured as median PE intensity in the indicated Karpas299 (left), A-498 (center) and HeLa (right) UGDH _KO cells expressing a sgRNA resistant UGDH cDNA (blue) or an empty vector (grey). f. Cellular uptake of DiI-HDL measured as median PE intensity in the indicated A-498 (left) and HeLa (right) UGDH _KO cells expressing a sgRNA resistant UGDH cDNA (blue) or an empty vector (grey). a, e, f, Lines or bars represent mean ± s.d.; a, e, f, n=3 biologically independent samples. Statistical significance determined by two-tailed unpaired t-tests.

Article Snippet: Antibodies to ACSL3 (Ab151959), human GPX4 (Ab41787), LDLR (52818), SCARB1(52629) were purchased from Abcam; Heparan Sulfate (370255-S) from AMSBio; IRP2 (37135S), SLC7A11 (12691S), TFRC (13208S), and β-tubulin (2146S) from Cell Signaling Technology; GAPDH (GTX627408) and UGDH (GTX104993) from GeneTex; AIFM2 (20866-1-AP) and PAPSS1 (14708-1-AP) from ProteinTech; mouse GPX4 (sc-166570) from Santa Cruz Biotechnology; and Chondroitin Sulfate (C8035) from Sigma.

Techniques: Expressing, Plasmid Preparation, Western Blot, Control, Flow Cytometry, Two Tailed Test