Journal: Cell Transplantation

Article Title: Histone Methyltransferase KMT2D Regulates H3K4 Methylation and is Involved in the Pathogenesis of Ovarian Cancer

doi: 10.1177/09636897211027521

Figure Lengend Snippet: (A) ESR1 expression in OV stage II–IV was higher than stage I and Fig. 1B low survival probability related to high ESR1 level from TCGA ( P = 0.022) (B) Immunohistochemical staining images (up) and scores (down) of KMT2D, ESR1 and H3K4me in tumor tissues of phase I ( n = 38), phase II ( n = 58), phase III ( n = 60) and phase IV ( n = 42) OV patients and adjacent tissues (including tumor samples and adjacent tissues.). (phase I vs phase II P < 0.05, phase II vs phase III P < 0.001, phase III vs phase IV N.S. , two-sided unpaired t -test). (C) The protein expression of KMT2D, ESR1, PTGS2, STAT3, VEGFR2 in tumor tissues of phase I ( n = 38), phase II ( n = 58), phase III ( n = 60) and phase IV ( n = 42) OV patients and adjacent tissues (including 198 tumor samples and 133 adjacent tissues). (phase I vs phase II P < 0.05, phase II vs phase III P < 0.001, phase III vs phase IV N.S. , two-sided unpaired t -test). (D) The binding of H3K4me and ESR1 was demonstrated using CHIP analysis and the relative expression of ESR1 was calculated via normalized to the Input group. Scale bar: 50 μm. * P < 0.05 vs adjacent tissues, ** P < 0.01 vs adjacent tissues, # P < 0.05 vs tumor (phase I), ## P < 0.01 vs tumor (phase I).

Article Snippet: The adenovirus vector for KMT2D interference, the plasmid for point mutation at H3K4 site, as well as the activator and inhibitor of KMT2D were constructed by GenePharma Co., Ltd (Shanghai, China), which were subsequently transfected into SKOV3 and HO-8910 cell lines for 48 h, respectively.

Techniques: Expressing, Immunohistochemical staining, Staining, Binding Assay

Journal: Cell Transplantation

Article Title: Histone Methyltransferase KMT2D Regulates H3K4 Methylation and is Involved in the Pathogenesis of Ovarian Cancer

doi: 10.1177/09636897211027521

Figure Lengend Snippet: (A) Expression of KMT2D, ESR1, H3K4me in IOSE80 and SKOV3 cell lines were determined using immunofluorescence and located by DAPI staining. (B) Expression of KMT2D, ESR1, FOXL2, PTGS2, and VEGFR2 detected by western blotting. The relative expression was calculated via normalization to the expression of actin. Scale bar: 20 μm. * P < 0.05 vs IOSE80, ** P < 0.01 vs IOSE80, # P < 0.05 vs model control group, ## P < 0.01 vs model control group.

Article Snippet: The adenovirus vector for KMT2D interference, the plasmid for point mutation at H3K4 site, as well as the activator and inhibitor of KMT2D were constructed by GenePharma Co., Ltd (Shanghai, China), which were subsequently transfected into SKOV3 and HO-8910 cell lines for 48 h, respectively.

Techniques: Expressing, Immunofluorescence, Staining, Western Blot

Journal: Cell Transplantation

Article Title: Histone Methyltransferase KMT2D Regulates H3K4 Methylation and is Involved in the Pathogenesis of Ovarian Cancer

doi: 10.1177/09636897211027521

Figure Lengend Snippet: (A) OV tumor size in mice was measured by ruler (mm 3 ). (B) Relative mRNA expression of ESR1, ELF3, and STAT3 determined using qRT-PCR and calculated via normalization to the mRNA level of GAPDH. (C) Expression of KMT2D, ESR1, H4K3me, PTGS2, and VEGFR2 detected by western blotting. The relative expression was calculated via normalization to the expression of actin. (D) The binding of H3K4me and ESR1 was demonstrated using CHIP analysis and the relative expression of ESR1 was calculated via normalized to the Input group. * P < 0.05 vs IOSE80, ** P < 0.01 vs IOSE80, # P < 0.05 vs model control group, ## P < 0.01 vs model control group.

Article Snippet: The adenovirus vector for KMT2D interference, the plasmid for point mutation at H3K4 site, as well as the activator and inhibitor of KMT2D were constructed by GenePharma Co., Ltd (Shanghai, China), which were subsequently transfected into SKOV3 and HO-8910 cell lines for 48 h, respectively.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Binding Assay

Journal: Oncogene

Article Title: Exposure of Barrett’s and esophageal adenocarcinoma cells to Bile acids activates EGFR-STAT3 signaling axis via induction of APE1

doi: 10.1038/s41388-018-0388-8

Figure Lengend Snippet: Immunoblot analysis of CPB ( A ), FLO-1 ( B ) and OE33 ( C ) cells pretreated with EGFR inhibitor (Gefitinib, 25 μM) followed by exposure to acidic (pH4) bile salts (100 μM). The samples were analyzed for the indicated proteins, β-actin was used as an internal control. ( D ) Immunoblot analysis of OE33 cells with EGFR-knockdown via EGFR siRNA followed by treatment with acidic (pH4) bile salts (100 μM) for 20 minutes and allowed to recover in complete media. The samples were collected at 3 and 6h post recovery and analyzed for the indicated proteins, β-actin was used as an internal control. Immunoprecipitation (IP) of APE1 ( E ), EGFR ( F ) and STAT3 ( G ) in OE33 cells treated with acidic (pH4) bile salts (100 μM) and immunoblotted for the indicated proteins. Results shown are representative of at least three independent experiments.

Article Snippet: Lentivirus particles expressing APE1 shRNA or control shRNA were produced by VectorBuilder Inc. (Santa Clara, CA, USA) and then used to transduce OE33 and CPB cells.

Techniques: Western Blot, Control, Knockdown, Immunoprecipitation

Journal: Frontiers in Genetics

Article Title: Long Non-coding RNA FGD5-AS1 Regulates Cancer Cell Proliferation and Chemoresistance in Gastric Cancer Through miR-153-3p/CITED2 Axis

doi: 10.3389/fgene.2020.00715

Figure Lengend Snippet: Effects of FGD5-AS1 inhibition on gastric cancer cell proliferation and 5-FU chemoresistance. (A) Two gastric cancer cell lines, SGC-7901 and MKN-28 were transduced with a lentivirus containing human lncRNA FGD5-AS1 sh_RNA (sh_FGD5AS1), or a lentivirus containing negative control shRNA (sh-NC). After transduction was stabilized, qRT-PCR was applied to compare FGD5-AS1 expression levels (* P < 0.05). (B) A 5-day MTT assay was used to assess in vitro proliferation rates between SGC-7901 and MKN-28 cells transduced with sh-NC and those transduced with sh-FGD5AS1 (** P < 0.05). (C) Lentiviral-transduced SGC-7901 and MKN-28 cells were treated with 0, 5, 10, 20, 50, or 100 μg/mL 5-FU for 24 h, followed by an MTT assay to compare 5-FU chemoresistance between those transduced with sh-NC and those transduced with sh-FGD5AS1 (** P < 0.05). (D) Lentiviral-transduced SGC-7901 cells were subcutaneously inoculated (1 × 10 6 /injection) into the flanks of 2-month-old female athymic mice for 5 weeks. The in vivo tumor volumes (mm 3 ) were compared between those transduced with sh-NC and those transduced with sh-FGD5AS1 (** P < 0.05). (E) After the in vivo tumorigenicity assay, SGC-7901 tumors were retrieved and compared directly.

Article Snippet: A lentivirus containing human lncRNA FGD5-AS1 short hairpin RNA (shRNA) (sh_FGD5AS1), and a lentivirus containing negative control shRNA (sh-NC) were purchased from SunBio (SunBio Medical Biotechnology, Shanghai, China).

Techniques: Inhibition, Transduction, Negative Control, shRNA, Quantitative RT-PCR, Expressing, MTT Assay, In Vitro, Injection, In Vivo, Tumorigenicity Assay