Journal: Pharmaceutics

Article Title: IDF-11774 Induces Cell Cycle Arrest and Apoptosis by Inhibiting HIF-1α in Gastric Cancer

doi: 10.3390/pharmaceutics15122772

Figure Lengend Snippet: Hypoxia-inducible factor-1 alpha (HIF-1α) knockdown suppresses the growth of gastric cancer cells via cell cycle arrest, apoptosis, and the mitogen-activated protein kinase (MAPK) pathway. ( A ) MKN45 cells were transfected with HIF-1α siRNA for 48 h and then lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, and E1), and β-actin. ( B ) MKN45 cells were transfected with HIF-1α siRNA for 48 h and nocodazole for 16 h, and then released into G1 for up to 40 h; nocodazole was replaced with fresh media. Samples were lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, E1, Cyclin-dependent kinase 4, p53, and p21), and β-actin. ( C ) MKN45 cells were transfected with HIF-1α siRNA for 48 h and then lysed and immunoblotted with HIF-1α, apoptosis-associated proteins (poly(ADP ribose) polymerase, cleaved caspase-3), and β-actin. ( D ) MKN45 cells were transfected with HIF-1α siRNAs for 48 h and then lysed and immunoblotted with HIF-1α, MAPK pathway proteins (phospho-extracellular signal-regulated protein kinases (ERK), ERK, phosphor-Jun N-terminal kinase (JNK), JNK, phosphor-p38, and p38), and β-actin.

Article Snippet: Approximately 20 μg of the protein extract was resolved via SDS-PAGE and analyzed via immunoblotting with primary antibodies against α-HIF-1α (Cell Signaling, Danvers, MA, USA, 3716), α-PARP (Cell Signaling, 9542), α-Caspase-3 (Cell Signaling, 9662), α-p44/42 MAPK (ERK1/2) (Cell Signaling, 4695), α-Phospho-p44/42 MAPK (Erk1/2) (Cell Signaling, 4370), α-p38 MAPK (Cell Signaling, 9212), α-Phospho-p38 MAPK (Cell Signaling, 4511), α-JNK2 (Cell Signaling, 9258), α-Phospho-SAPK/JNK (Cell Signaling, 4668), α-Cyclin B1 (Santa Cruz, Dallas, TX, USA, sc-752), α-Cyclin D1 (Cell Signaling, 2978), α-Cyclin E1 (Cell Signaling, 4129), α-CDK4 (Cell Signaling, 12790), α-p53 (Millipore, Burlington, MA, USA, 05-224), α-p21 (Cell Signaling, 2947), α-TBP (Abcam, Cambridge, UK, ab818), α-β-actin (Santa Cruz, sc-47778), and α-alpha-tubulin (Abcam, ab7291).

Techniques: Knockdown, Transfection

Journal: Pharmaceutics

Article Title: IDF-11774 Induces Cell Cycle Arrest and Apoptosis by Inhibiting HIF-1α in Gastric Cancer

doi: 10.3390/pharmaceutics15122772

Figure Lengend Snippet: IDF-11774 induces cell cycle arrest and apoptosis by downregulating hypoxia-inducible factor-1 alpha (HIF-1α) in gastric cancer cells. ( A ) MKN45 and MKN74 cells were treated with 30 μM IDF-11774 for 4 h and then lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, and E1), and β-actin. ( B ) MKN45 and MKN74 cells were treated with 30 μM IDF-11774 for 4 h and nocodazole for 16 h. Thereafter, the cells were released into G1 for up to 40 h; nocodazole was replaced with fresh media. Samples were lysed and immunoblotted with HIF-1α, cell cycle arrest-associated proteins (Cyclin B1, D1, E1, cyclin-dependent kinase 4, p53, and p21), and β-actin. ( C ) MKN45 and MKN74 cells were treated with 30 μM IDF-11774 for 4 h and then lysed and immunoblotted with HIF-1α, apoptosis-associated proteins (poly(ADP ribose) polymerase, cleaved caspase-3), and β-actin. ( D ) MKN45 and MKN74 cells were treated with 30 μM IDF-11774 for 4 h and then lysed and immunoblotted with HIF-1α, mitogen-activated protein kinase pathway proteins (phospho-extracellular signal-regulated protein kinases (ERK), ERK, phosphor-Jun N-terminal kinase (JNK), JNK, phosphor-p38, and p38), and β-actin.

Article Snippet: Approximately 20 μg of the protein extract was resolved via SDS-PAGE and analyzed via immunoblotting with primary antibodies against α-HIF-1α (Cell Signaling, Danvers, MA, USA, 3716), α-PARP (Cell Signaling, 9542), α-Caspase-3 (Cell Signaling, 9662), α-p44/42 MAPK (ERK1/2) (Cell Signaling, 4695), α-Phospho-p44/42 MAPK (Erk1/2) (Cell Signaling, 4370), α-p38 MAPK (Cell Signaling, 9212), α-Phospho-p38 MAPK (Cell Signaling, 4511), α-JNK2 (Cell Signaling, 9258), α-Phospho-SAPK/JNK (Cell Signaling, 4668), α-Cyclin B1 (Santa Cruz, Dallas, TX, USA, sc-752), α-Cyclin D1 (Cell Signaling, 2978), α-Cyclin E1 (Cell Signaling, 4129), α-CDK4 (Cell Signaling, 12790), α-p53 (Millipore, Burlington, MA, USA, 05-224), α-p21 (Cell Signaling, 2947), α-TBP (Abcam, Cambridge, UK, ab818), α-β-actin (Santa Cruz, sc-47778), and α-alpha-tubulin (Abcam, ab7291).

Techniques: