fth1 sirna  (Santa Cruz Biotechnology)

Journal: Frontiers in Pharmacology

Article Title: Anticancer activity of RM-3-22: a TAZQ-based hydroxamic acid derivative targeting NSCLC in vitro and in vivo

doi: 10.3389/fphar.2025.1544666

Figure Lengend Snippet: RM-3-22 induces the upregulation of FTH1, a tumor suppressor of NSCLC, which is positively correlated with apoptosis and autophagy pathway. (A) Western blot analysis shows the increased expression of FTH1. Data represent the mean ± SD of three independent experiments (one-way ANOVA; *p < 0.05, **p < 0.01, and ***p < 0.001). (B) qRT-PCR of FTH1 shows an increase in the mRNA level due to RM-3-22 exposure, both in a concentration-dependent manner. Data represent the mean ± SD of three independent experiments (one-way ANOVA; *p < 0.05, **p < 0.01, and ***p < 0.001). (C) Protein–ligand interaction diagram; (a) crystal structure of FTH-1 (PDB: 6J4A) interacting with ligand RM-3-22. (b) 2D molecular interaction of FTH-1 with RM22, docked using AutoDock Vina. Projected bonds are shown. Docking score -8.1. (D) Kaplan–Meier plotter survival analysis for patients with expression of FTH1. Survival analysis was performed in KM plotter using publicly available microarray data that are available within the database. A log-rank p-value ≤0.05 was considered significant. HR, hazard ratio. The red curve represents high expression, and the black curve represents low expression. OS)of the patients increases with high expression of FTH1 in lung cancer. (E) The FTH1 transcript level seems to decrease slightly during LUAD and lung squamous cell carcinoma (LUSC) progression in the patients (UALCAN database, TGCA sample), which indicates the tumor-suppressive nature of FTH1 in lung cancer. (F) PPI network of FTH1, autophagy, and apoptosis-related proteins constructed in Cytoscape based on interaction information derived from the STRING database. The network contains 10 nodes (proteins) and 37 edges (interactions). PPI showed the interaction of FTH1 with SQSTM1 and MAP1LC3B and FTH1 with CYCS (cytochrome C). For this specific interaction, edge color is specified as red. (G) Gene correlation analysis exhibited a positive correlation of FTH1 with MAP1LC3B and pro-apoptotic protein BAX and a negative correlation with anti-apoptotic protein Bcl2 at a significance level (*p < 0.05) (data from TCGA, cbiojune.en). (H) FTH1 knockdown by siRNA in A549 cells. Data represent the mean ± SD of three independent experiments (t-test; *p < 0.05, **p < 0.01, and ***p < 0.001). (I) Migration efficiency was higher in FTH1-downregulated cells treated with RM-3-22 compared to normal A549 cells solely treated with RM-3-22. (J) FTH1 knockdown led to the reversal of nuclear fragmentation, which is accompanied by RM-3-22. (K) Schematic representation showing the regulation of FTH1 by RM-3-22.

Article Snippet: Human-specific FTH1 siRNA was transfected into A549 cells in accordance with the manufacturer’s instructions (Santa-Cruz, sc-40575).

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Concentration Assay, Microarray, Construct, Derivative Assay, Knockdown, Migration

Journal: Frontiers in Pharmacology

Article Title: Anticancer activity of RM-3-22: a TAZQ-based hydroxamic acid derivative targeting NSCLC in vitro and in vivo

doi: 10.3389/fphar.2025.1544666

Figure Lengend Snippet: RM-3-22 suppresses tumor growth and induces autophagy, apoptosis, cell-cycle arrest, and FTH1 upregulation. (A) Tumor volume reduced in the treated group after treatment of RM-3-22 (DC, disease control; TG, treated group). Tumor sizes were measured every 2 days using the formula tumor volume= (width) 2 x length/2. (B) Representative photomicrographs of hematoxylin and eosin staining showed disrupted tissue structure and damaged nuclei in RM-3-22-treated tissue. Scale bar = 100 μm (the box indicates the close-up view of disrupted structures; scale bar = 50 μm). (C) IHC of MAP1LC3B protein expression from DC and TG of mice. Scale bar = 100 μm (the box indicates the close-up view; scale bar = 50 μm). Data represent the mean ± SD of three independent experiments (t-test; *p < 0.05, **p < 0.01, and ***p < 0.001). (D) Western blot analysis was used to calculate the expression levels of MAP1LC3B, FTH1, cyclin B 1, and PARP-1 in tumor samples. Data represent the mean ± SD of three independent experiments (*p < 0.05, **p < 0.01, and ***p < 0.001). Schematic representation is presented in (E) .

Article Snippet: Human-specific FTH1 siRNA was transfected into A549 cells in accordance with the manufacturer’s instructions (Santa-Cruz, sc-40575).

Techniques: Control, Staining, Expressing, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Anticancer activity of RM-3-22: a TAZQ-based hydroxamic acid derivative targeting NSCLC in vitro and in vivo

doi: 10.3389/fphar.2025.1544666

Figure Lengend Snippet: Summary of the mechanism of action after RM-3-22 treatment. The diagram indicates that RM-3-22 is responsible for the inhibition of the PI3K/Akt/mTOR pathway, which leads to the activation of the autophagy pathway, marked by the induction of regulatory makers of autophagy, MAP1LC3B II and Beclin1, and also the degradation of p62. The induction of the autophagy pathway further induces the apoptosis pathway, confirmed by the translocation of BAX from the cytosol to the nucleus, and subsequently the cleavage of PARP-1 and caspase 3. RM-3-22-induced autophagy is further responsible for the G2/M cellcycle arrest, and this is confirmed by the destruction of cyclin B1 and CDC2 in a dose-dependent way. Apart from that, RM-3-22 activates FTH1-related tumor suppression in NSCLC.

Article Snippet: Human-specific FTH1 siRNA was transfected into A549 cells in accordance with the manufacturer’s instructions (Santa-Cruz, sc-40575).

Techniques: Inhibition, Activation Assay, Translocation Assay

Journal: bioRxiv

Article Title: TvAP65 in Trichomonas vaginalis Promotes HPV Infection by Interacting with Host Molecules

doi: 10.1101/2024.09.27.615334

Figure Lengend Snippet: “Control” represents HaCaT cells that have not received any treatment; “HPV” represents HaCaT cells infected with HPV; “TV+HPV” represents HaCaT cells infected with both T. vaginalis and HPV; “SPCS1”, “FTH1”, “ATP5MC3”, “IGFBP7”, “PMEPA1”, “REEP5”, “BNIP3”, “C4orf3”, “C6orf89”, “GRAMD1A”, “TNFRSF12A”, and “PKM” represent HaCaT cells infected with both T. vaginalis and HPV under low-expression of TvAP65 interaction molecules. A The HPV infection in HaCaT cells was observed with fluorescence microscope after different genes were down-regulated. B and C The HPV infection rate in HaCaT cells was detected by flow cytometry after different genes were down-regulated. “ns” indicates no statistical difference, and “***” indicates P <0.001.

Article Snippet: 10 μL of 10 μM siRNA for CD151 or HSPG2 genes, or 20 μM siRNA for FTH1, SPCS1, ATP5MC3, IGFBP7, PMEPA1, REEP5, BNIP3, C4orf3, C6orf89, GRAMD1A, TNFRSF12A, or PKM genes were mixed with 250 μL opti-MEM I reduced-serum medium (Thermo Fisher Scientific) and gently pipetted.

Techniques: Control, Infection, Expressing, Fluorescence, Microscopy, Flow Cytometry

Journal: bioRxiv

Article Title: TvAP65 in Trichomonas vaginalis Promotes HPV Infection by Interacting with Host Molecules

doi: 10.1101/2024.09.27.615334

Figure Lengend Snippet: “Control” represents untreated HaCaT cells, “TV” represents HaCaT cells infected with T. vaginalis , and “SPCS1-siRNA+TV” represents that HaCaT cells infected with T. vaginalis under low-expression of SPCS1. “NC-siRNA+TV” and “Lipo2000+TV” served as the control group of NC siRNA and Lipofectamine 2000 respectively. A qPCR was used to detect the mRNA levels of HSPG2 and CD151 in HaCaT cells; B WB was used to detect the protein expression of HSPG2 and CD151 in HaCaT cells. C Image J software was used to analyze the protein expression levels of HSPG2 and CD151. “ns” indicates no statistical difference, and “***” indicates P <0.001.

Article Snippet: 10 μL of 10 μM siRNA for CD151 or HSPG2 genes, or 20 μM siRNA for FTH1, SPCS1, ATP5MC3, IGFBP7, PMEPA1, REEP5, BNIP3, C4orf3, C6orf89, GRAMD1A, TNFRSF12A, or PKM genes were mixed with 250 μL opti-MEM I reduced-serum medium (Thermo Fisher Scientific) and gently pipetted.

Techniques: Control, Infection, Expressing, Software

Journal: bioRxiv

Article Title: TvAP65 in Trichomonas vaginalis Promotes HPV Infection by Interacting with Host Molecules

doi: 10.1101/2024.09.27.615334

Figure Lengend Snippet: “Control” represents untreated HaCaT cells; “HPV” represents HaCaT cells infected with HPV; “TV+HPV” represents HaCaT cells infected with both T. vaginalis and HPV; “TvAP65”, “SPCS1”, and “SPCS1/TvAP65” represent HaCaT cells infected with both T. vaginalis and HPV under low-expression of the TvAP65, SPCS1, or SPCS1/TvAP65 genes, respectively. A The HPV infection in HaCaT cells was observed with fluorescence microscopy after low-expression of different genes; B and C The HPV infection rate in HaCaT cells was detected by flow cytometry after after different genes were down-regulated. “ns” indicates no statistical difference, “**” indicates P <0.01, and “***” indicates P <0.001.

Article Snippet: 10 μL of 10 μM siRNA for CD151 or HSPG2 genes, or 20 μM siRNA for FTH1, SPCS1, ATP5MC3, IGFBP7, PMEPA1, REEP5, BNIP3, C4orf3, C6orf89, GRAMD1A, TNFRSF12A, or PKM genes were mixed with 250 μL opti-MEM I reduced-serum medium (Thermo Fisher Scientific) and gently pipetted.

Techniques: Control, Infection, Expressing, Fluorescence, Microscopy, Flow Cytometry

Journal: bioRxiv

Article Title: TvAP65 in Trichomonas vaginalis Promotes HPV Infection by Interacting with Host Molecules

doi: 10.1101/2024.09.27.615334

Figure Lengend Snippet: “Control” represents HaCaT cells that have not received any treatment; “HPV” represents HaCaT cells infected with HPV; “TV+HPV” represents HaCaT cells infected with both T. vaginalis and HPV; “SPCS1”, “FTH1”, “ATP5MC3”, “IGFBP7”, “PMEPA1”, “REEP5”, “BNIP3”, “C4orf3”, “C6orf89”, “GRAMD1A”, “TNFRSF12A”, and “PKM” represent HaCaT cells infected with both T. vaginalis and HPV under low-expression of TvAP65 interaction molecules. A The HPV infection in HaCaT cells was observed with fluorescence microscope after different genes were down-regulated. B and C The HPV infection rate in HaCaT cells was detected by flow cytometry after different genes were down-regulated. “ns” indicates no statistical difference, and “***” indicates P <0.001.

Article Snippet: 10 μL of 10 μM siRNA for CD151 or HSPG2 genes, or 20 μM siRNA for FTH1, SPCS1, ATP5MC3, IGFBP7, PMEPA1, REEP5, BNIP3, C4orf3, C6orf89, GRAMD1A, TNFRSF12A, or PKM genes were mixed with 250 μL opti-MEM I reduced-serum medium (Thermo Fisher Scientific) and gently pipetted.

Techniques: Control, Infection, Expressing, Fluorescence, Microscopy, Flow Cytometry

Journal: Emerging microbes & infections

Article Title: Enterovirus-A71 preferentially infects and replicates in human motor neurons, inducing neurodegeneration by ferroptosis.

doi: 10.1080/22221751.2024.2382235

Figure Lengend Snippet: Figure 4. Ferroptosis in human motor neurons after EV-A71 infection. A. Expression levels of FTL and FTH1 after infection in each population. B. Expression levels of FTL and FTH1 after infection in each population with high viral RNA load (High) and low viral RNA load (Low). C. Free irons within the cells were measured by Ferro-Orange. D. Number of cells that were positive for Ferro- Orange in the EV-A71-infected and mock-infected cultures. E. Ferro-Orange intensity in each cell at 6 h after EV-A71 or mock infec tion. n > 1900 cells from 3 cultures, Mann-Whitney Test, ****p < 0.0001. F. Lipid peroxidation measured by BODIPY C11 581/ 591. G. Number of cells displaying oxidized C11 in EV-A71-infected and mock cultures. H. Mean oxidized C11 intensity in each cell at 24 h after EV-A71 or mock infection. n > 2300 cells from 3 cultures, Mann-Whitney Test, ****p < 0.0001.

Article Snippet: The siRNAs used were: SCARB2 (Dharmacon, #012087), FTL (SCBT, sc40577), FTH1 (SCBT, sc-40575).

Techniques: Infection, Expressing, MANN-WHITNEY