Journal: Frontiers in Oncology

Article Title: Next-Generation Sequencing Analysis of Gastric Cancer Identifies the Leukemia Inhibitory Factor Receptor as a Driving Factor in Gastric Cancer Progression and as a Predictor of Poor Prognosis

doi: 10.3389/fonc.2022.939969

Figure Lengend Snippet: LIFR is a negative prognostic factor for survival of a patient with GC with carcinomatosis. The expression of LIFR and LIF was examined in surgical samples from non-neoplastic and gastric adenocarcinoma mucosa obtained by patients with GC underwent surgery for GC treatment. Data shown are follows: Gene expression of LIFR (Log2) (A) in non-neoplastic vs. neoplastic mucosa and (B) in non-neoplastic and gastric adenocarcinoma w/o carcinomatosis vs. adenocarcinoma with carcinomatosis. (C) Relative mRNA expression of LIF. (D) H&E staining of non-neoplastic mucosa (magnification, ×20). (E) IHC staining of non-neoplastic mucosa (magnification, ×20, ×40, and ×100). (F) (hematoxylin and eosin) H&E staining of gastric adenocarcinoma mucosa (magnification, ×20). (G) IHC staining of IHC staining of gastric adenocarcinoma mucosa (magnification, ×20, ×40, and ×100). * represents statistical significance versus Non neoplastic tissue.

Article Snippet: In another experimental setting, on day 3, cells were triggered with LIF (10 ng/ml) plus LIFR antagonist, EC359 (25, 50, 100, and 1,000 nM) (MedChemExpress, NJ 08852, USA), and cell proliferation was assessed as mentioned above.

Techniques: Expressing, Staining, Immunohistochemistry

Journal: Frontiers in Oncology

Article Title: Next-Generation Sequencing Analysis of Gastric Cancer Identifies the Leukemia Inhibitory Factor Receptor as a Driving Factor in Gastric Cancer Progression and as a Predictor of Poor Prognosis

doi: 10.3389/fonc.2022.939969

Figure Lengend Snippet: LIFR activation promotes cell proliferation and EMT in MKN45 cells. Relative mRNA expression (A) LIFR and (B) LIF in CG cell lines. (C) IHC staining of LIFR in MNK45 cell lines on the left untreated and on the right triggered with LIF (10 ng/ml; magnification, ×100). MKN45 cells were serum-starved and primed with LIF (0.5, 5, and 10 ng/ml). Data shown are as follows: (D) dose–response curve of LIF (0.5, 5, and 10 ng/ml) determined using MTS assay on MKN45 cells. Each value is expressed relative to those of non-treated (NT), which are arbitrarily settled to 1. Results are the mean ± SEM of 10 samples per group. Relative mRNA expression of (E) the proliferation marker C-Myc and EMT markers (F) E-cadherin, (G) vimentin, and (H) Snal-1. Each value is normalized to Gapdh and is expressed relative to those of positive controls, which are arbitrarily settled to 1. Results are the mean ± SEM of five samples per group (* represents statistical significance versus NT, and # versus LIF, p < 0.05).

Article Snippet: In another experimental setting, on day 3, cells were triggered with LIF (10 ng/ml) plus LIFR antagonist, EC359 (25, 50, 100, and 1,000 nM) (MedChemExpress, NJ 08852, USA), and cell proliferation was assessed as mentioned above.

Techniques: Activation Assay, Expressing, Immunohistochemistry, MTS Assay, Marker

Journal: Frontiers in Oncology

Article Title: Next-Generation Sequencing Analysis of Gastric Cancer Identifies the Leukemia Inhibitory Factor Receptor as a Driving Factor in Gastric Cancer Progression and as a Predictor of Poor Prognosis

doi: 10.3389/fonc.2022.939969

Figure Lengend Snippet: LIFR antagonist EC359 hinders cell cycle progression, increases apoptosis rate in MKN45 cells and inhibits EMT process. (A) Dose–response curve of EC359 (25, 50, 100, and 1,000 nM) determined using MTS assay on MKN45 cells (n = 10). MKN45 cells were serum-starved and triggered with LIF (10 ng/ml), EC359 (100 nM), and LIF + EC359 for 48 h. Cell cycle phase analysis was performed by Ki-67/DAPI staining through IC-FACS. Data shown are follows: percentage of (B) from left to right cell in G0-G1 cell cycle phases, S-G2-M cell cycle phases, and ratio between % G0-G1 and % S-G2-M. (C) Percentage of apoptotic cells. (D) Representative IC-FACS showed cell cycle fraction and apoptosis rate in NT, LIF (10 ng/ml), EC359 (25 nM), and LIF + EC359. Results are the mean ± SEM of three samples for group (* represents statistical significance versus NT, and # versus LIF, p < 0.05). Relative mRNA expression of (E) the proliferation marker C-Myc and EMT markers (F) E-Cadherin, (G) Snal-1, and (H) vimentin. Each value is normalized to Gapdh and is expressed relative to those of positive controls, which are arbitrarily settled to 1. Results are the mean ± SEM of five samples per group (* represents statistical significance versus NT, and # versus LIF, p < 0.05).

Article Snippet: In another experimental setting, on day 3, cells were triggered with LIF (10 ng/ml) plus LIFR antagonist, EC359 (25, 50, 100, and 1,000 nM) (MedChemExpress, NJ 08852, USA), and cell proliferation was assessed as mentioned above.

Techniques: MTS Assay, Staining, Expressing, Marker

Journal: Frontiers in Oncology

Article Title: Next-Generation Sequencing Analysis of Gastric Cancer Identifies the Leukemia Inhibitory Factor Receptor as a Driving Factor in Gastric Cancer Progression and as a Predictor of Poor Prognosis

doi: 10.3389/fonc.2022.939969

Figure Lengend Snippet: Analysis of JAK-STAT signaling pathway. Representative Western blot analysis of (A) LIFR, JAK1 and phospho-JAK1, STAT3 and phospho-STAT3, and proteins in MKN45 exposed to LIF (10 nM) alone or in combination with EC359 (25 nM and 100 nM) for 20 min. GAPDH was used as loading control. (B) Densitometric analysis demonstrating LIFR expression, phospho-JAK1/JAK1, and phospho-STAT3/STAT3 ratio. The blot shown is representative of another one showing the same pattern. (* represents statistical significance versus NT, and # versus LIF, p < 0.05).

Article Snippet: In another experimental setting, on day 3, cells were triggered with LIF (10 ng/ml) plus LIFR antagonist, EC359 (25, 50, 100, and 1,000 nM) (MedChemExpress, NJ 08852, USA), and cell proliferation was assessed as mentioned above.

Techniques: Western Blot, Expressing

Journal: Frontiers in Oncology

Article Title: Next-Generation Sequencing Analysis of Gastric Cancer Identifies the Leukemia Inhibitory Factor Receptor as a Driving Factor in Gastric Cancer Progression and as a Predictor of Poor Prognosis

doi: 10.3389/fonc.2022.939969

Figure Lengend Snippet: LIFR antagonism inhibits MKN45. (A, B) A scratch wound healing assay is shown. MKN45 cell monolayers were scraped in a straight line using a p200 pipette tip to create a “scratch”, and then, they are left untreated or primed with LIF (10 ng/ml), EC359 (100 nM), and LIF + EC359. The wound generated was imaged at 0, 24, and 48 h of incubation with the indicated compounds. (A) The images show cell migration at the three times point indicated. (B) Images obtained points were analysed measuring scraped area and its closure vs. the first time point at 0 h. Results are the mean ± SEM of three samples per group. (C) Cell adhesion to peritoneum. Experiment was conducted in quintuplicate (* represents statistical significance versus NT, and # versus LIF, p < 0.05).

Article Snippet: In another experimental setting, on day 3, cells were triggered with LIF (10 ng/ml) plus LIFR antagonist, EC359 (25, 50, 100, and 1,000 nM) (MedChemExpress, NJ 08852, USA), and cell proliferation was assessed as mentioned above.

Techniques: Wound Healing Assay, Transferring, Generated, Incubation, Migration