Journal: Experimental and Therapeutic Medicine

Article Title: Effect of nicotine on cholesterol gallstone formation in C57BL/6J mice fed on a lithogenic diet

doi: 10.3892/etm.2023.11783

Figure Lengend Snippet: Effect of nicotine on cholesterol metabolism by regulating in the gallbladder epithelial tissues. (A) mRNA levels of bile acid metabolism in the gallbladder epithelial tissues of mice injected with nicotine was measured by quantitative polymerase chain reaction (n=3). (B) Representative western blot analyses and (C-E) immunohistochemical staining of FXR and megalin (magnification, x100). Data are expressed as the mean ± standard deviation of the mean (n=10 mice per group). * P<0.05 vs. the ND group; # P<0.05 vs. the LD group. ND, normal diet; LD, lithogenic diet; H, 6.6 mg/kg/2 days nicotine; L, 1.1 mg/kg/2 days nicotine; NPC1L1, NPC1 like intracellular cholesterol transporter 1; ABC, ATP-binding cassette; SR-BI, scavenger receptor class B member 1; FXR, farnesoid X receptor.

Article Snippet: The human gallbladder epithelial cell line GBEC were purchased from iCell Bioscience Inc. and were cultured in low-glucose DMEM supplemented with 2 mM glutamine, 1% MEM non-essential amino acids solution, 1% MEM vitamin solution, 7.5% FBS and P/S (all from Gibco; Thermo Fisher Scientific, Inc.).

Techniques: Injection, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemical staining, Staining, Standard Deviation, Binding Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Effect of nicotine on cholesterol gallstone formation in C57BL/6J mice fed on a lithogenic diet

doi: 10.3892/etm.2023.11783

Figure Lengend Snippet: Effect of nicotine on the regulation of FXR/megalin/cubilin in the gallbladder epithelial cells. (A) GBEC cells were treated with 2 µM nicotine for 48 h, then total cell lysates (40 µg) from GBECs were subjected to western blot analysis. (B) Immunofluorescence for megalin in confluent GBECs treated with nicotine for 48 h in the presence of FXR agonist CDCA (10 µM; magnification, x100). * P<0.05 vs. control; # P<0.05 vs. the CDCA. FXR, farnesoid X receptor; CDCA, chenodeoxycholic acid.

Article Snippet: The human gallbladder epithelial cell line GBEC were purchased from iCell Bioscience Inc. and were cultured in low-glucose DMEM supplemented with 2 mM glutamine, 1% MEM non-essential amino acids solution, 1% MEM vitamin solution, 7.5% FBS and P/S (all from Gibco; Thermo Fisher Scientific, Inc.).

Techniques: Western Blot, Immunofluorescence, Control

Journal: Oncology Letters

Article Title: Long non-coding RNA TMPO-AS1 promotes cell proliferation, migration, invasion and epithelial-to-mesenchymal transition in gallbladder carcinoma by regulating the microRNA-1179/E2F2 axis

doi: 10.3892/ol.2021.13116

Figure Lengend Snippet: TMPO-AS1 knockdown inhibits the proliferation, migration, invasion and EMT of GBC cells. (A) Reverse transcription-quantitative PCR analysis was performed to detect TMPO-AS1 expression in GBC cells transfected with sh-TMPO-AS1. (B) The Cell Counting Kit-8 assay was performed to assess the viability of GBC cells transfected with sh-TMPO-AS1 at the indicated times. (C) The colony formation assay was performed to assess the proliferation of GBC cells transfected with sh-TMPO-AS1. (D) The wound healing assay was performed to assess the migratory ability of GBC cells transfected with sh-TMPO-AS1. (E) Transwell migration and invasion assays were performed to assess the migratory and invasive abilities of GBC cells transfected with sh-TMPO-AS1. (F) The immunofluorescence assay was performed to detect α-SMA protein expression in GBC cells transfected with sh-TMPO-AS1. (G) Western blot analysis was performed to detect the protein expression levels of EMT markers, including E-cadherin, vimentin and α-SMA, in GBC cells transfected with sh-TMPO-AS1. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01 vs. sh-NC. TMPO-AS1, thymopoietin antisense transcript 1; EMT, epithelial-to-mesenchymal transition; GBC, gallbladder carcinoma; sh, short hairpin; SMA, smooth muscle actin; NC, negative control; OD, optical density.

Article Snippet: The GBC cell lines, SGC-996, GBC-SD, EH-GB1 and NOZ, and the human gallbladder epithelial cell line, H69 were purchased from the American Type Culture Collection.

Techniques: Knockdown, Migration, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Transfection, Cell Counting, Colony Assay, Wound Healing Assay, Immunofluorescence, Western Blot, Negative Control

Journal: Oncology Letters

Article Title: Long non-coding RNA TMPO-AS1 promotes cell proliferation, migration, invasion and epithelial-to-mesenchymal transition in gallbladder carcinoma by regulating the microRNA-1179/E2F2 axis

doi: 10.3892/ol.2021.13116

Figure Lengend Snippet: Overexpression of miR-1179 inhibits the proliferation, migration, invasion and EMT of GBC cells. (A) Reverse transcription-quantitative PCR analysis was performed to detect miR-1179 expression in GBC cells transfected with miR-1179 mimics. (B) The Cell Counting Kit-8 assay was performed to assess the viability of GBC cells transfected with miR-1179 mimics at the indicated times. (C) The colony formation assay was performed to assess the proliferation of GBC cells transfected with miR-1179 mimics. (D) The wound healing assay was performed to assess the migratory ability of GBC cells transfected with miR-1179 mimics. (E) The immunofluorescence assay was performed to detect α-SMA protein expression in GBC cells transfected with miR-1179 mimics. (F) The Transwell migration and invasion assays were performed to assess the migratory and invasive abilities of GBC cells transfected with miR-1179 mimics. (G) Western blot analysis was performed to detect the protein expression levels of the EMT markers, including E-cadherin, vimentin and α-SMA. Data are presented as the mean ± SD (n=3). *P<0.05, **P<0.01 vs. NC mimic. miR, microRNA; EMT, epithelial-to-mesenchymal transition; GBC, gallbladder carcinoma; TMPO-AS1, thymopoietin antisense transcript 1; SMA, smooth muscle actin; NC, negative control; OD, optical density.

Article Snippet: The GBC cell lines, SGC-996, GBC-SD, EH-GB1 and NOZ, and the human gallbladder epithelial cell line, H69 were purchased from the American Type Culture Collection.

Techniques: Over Expression, Migration, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Transfection, Cell Counting, Colony Assay, Wound Healing Assay, Immunofluorescence, Western Blot, Negative Control

Journal: Oncology Letters

Article Title: Long non-coding RNA TMPO-AS1 promotes cell proliferation, migration, invasion and epithelial-to-mesenchymal transition in gallbladder carcinoma by regulating the microRNA-1179/E2F2 axis

doi: 10.3892/ol.2021.13116

Figure Lengend Snippet: TMPO-AS1 regulates the proliferation, migration, invasion and EMT of GBC cells by modulating the miR-1179/E2F2 pathway. (A) Reverse transcription-quantitative PCR analysis was performed to detect the expression levels of TMPO-AS1, miR-1179 and E2F2 in GBC cells following transfection. (B) The Cell Counting Kit-8 assay was performed to assess the viability of GBC cells following transfection at the indicated times. (C) The colony formation assay was performed to assess the proliferation of GBC cells following transfection. (D) The wound healing assay was performed to assess the migration of GBC cells following transfection. (E) The Transwell migration and invasion assays were performed to assess the migratory and invasive abilities of GBC cells following transfection. (F) The immunofluorescence assay was performed to detect α-SMA protein expression in GBC cells following transfection. (G) Western blot analysis was performed to detect the protein expression levels of EMT markers, including E-cadherin and vimentin. Data are presented as the mean ± SD (n=3). **P<0.01 and ***P<0.001 vs. sh-NC + NC inhibitor; # P<0.05, ## P<0.01 and ### P<0.001 vs. sh-TMPO-AS1 + NC inhibitor; $ P<0.05, $$ P<0.01 and $$$ P<0.001 vs. sh-TMPO-AS1 + miR-1179 inhibitor. TMPO-AS1, thymopoietin antisense transcript 1; EMT, epithelial-to-mesenchymal transition; miR, microRNA; E2F2, E2F transcription factor 2; GBC, gallbladder carcinoma; sh, short hairpin; NC,negative control; OD, optical density.

Article Snippet: The GBC cell lines, SGC-996, GBC-SD, EH-GB1 and NOZ, and the human gallbladder epithelial cell line, H69 were purchased from the American Type Culture Collection.

Techniques: Migration, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Transfection, Cell Counting, Colony Assay, Wound Healing Assay, Immunofluorescence, Western Blot, Negative Control