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Seminario Biologia Molecular

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Seminario Biologia Molecular

  1. 1. SEBASTIÁN HERNÁNDEZ CANO 3RD SEMESTER MOLECULAR BIOLOGY 1 1 1
  2. 2. Breast cancer Reffers to a pathogenic growth of cells around the breast area that lose the ability to die 2 2 2
  3. 3. KDM4Asignaling pathway Induces cell growth, proliferation and survival by regulating (+) DNA repair processes 3 3 3 HTTPS://WWW.GOOGLE.COM/URL? SA=I&URL=HTTPS%3A%2F%2FLINK.SPRINGER.COM%2FARTICLE%2F10.1007%2FS00018- 019-03289- W&PSIG=AOVVAW33FJG0_CETROGROUJ6JK2I&UST=1675647277961000&SOURCE=IMAG ES&CD=VFE&VED=0CBAQJRXQFWOTCNIX9NEE_FWCFQAAAAADAAAAABAE
  4. 4. General Objective General Objective "Evaluate epigenetic regulator KDM4A activates Notch1-NICD-dependent signaling to drive tumorigenesis and metastasis in breast cancer " 4 4 4
  5. 5. Materials and Methods INMUNOHISTOCHEMICAL STAINING RNA ISOLATION AND QUANTITATIVE RT-PCR 5 5 5 50 samples were extracted and analyzed with GT vision III staining kit Then, staining was classified as none (0), weak (1), moderate (2) and strong (3) both by intensity and covered area mRNA was isolated and transcribed to cDNA by reverse transcriptase, then PCR was performed (RT-PCR process)
  6. 6. inmmunoreactions antigen-antibody were performed for KDM4A, Notch1, CCND3 and β-actin. gels were performed using electrophoresis Materialsandmethods 6 6 6 Western Blot analysis Proteins of interest bound to DNA were marked with antibodies (H3K27ac and H3K9me3). the DNA was then purified and qPCR was performed ChIP-qPCR assay https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.bio-rad.com%2Fen-us%2Fapplications-technologies%2Fprotein-electrophoresis- methods&psig=AOvVaw2eMLzREzOM6SyYWpkiPaEG&ust=1675639707073000&source=images&cd=vfe&ved=0CBAQjRxqFwoTCKi5vr6C_fwCFQA AAAAdAAAAABAE https://www.google.com/url?sa=i&url=https%3A%2F%2Fwww.cellsignal.com%2Fapplications%2Fchip-and- chip-seq%2Fregulation-expression-in-cell-and- tissue&psig=AOvVaw2ZZtAvhnjdjAtJX9TlhYA2&ust=1675641402433000&source=images&cd=vfe&ved=0 CBAQjRxqFwoTCNjBheaI_fwCFQAAAAAdAAAAABAE
  7. 7. KDM4A SILENCER RNA NOTCH1 SIGNALING IN KNOCKDOWNS LOW/HIGH CLASSIFICATION Results 7 7 7 Low High
  8. 8. KDM4A KNOCKDOWN (F) AND OVEREXPRESSION (G) KDM4A INCREASES CELL VIABILITY AND PROLIFERATION NOTCH 1 INHIBITION COULD SUPRESS DOWNSTREAM GENES Results 8 8 8
  9. 9. Author Statement Status S.Z. Cui In prostate cancer, USP1 is a deubiquitinase that regulates KDM4A K48-linked deubiquitin and stability to drive tumor cells proliferation and enzalutamide resistance Agree J. Lin Oncogene APOL1 promotes proliferation and inhibits apoptosis via activating NOTCH1 signaling pathway in pancreatic cancer Agree M.A.J. Morgan, A. Shilatifard histone modifications exert essential functions in cell fate determination, terminal differentiation and X inactivation Agree Discussion 9 9 9
  10. 10. Conclusions Asserting molecular pathways in pathological events could help provide therapeutic targets such as KDM4A or Notch1 learning about the tissue behavior behind protein overexpression/knockdown might help establish the reason to certain pathologies and develop more specific and efficient treatments 10 10 10
  11. 11. Epigenetic regulator KDM4A activates Notch1- NICD-dependent signaling to drive tumorigenesis and metastasis in breast cancer NOTCH1 KDM4A is a histone demethylase that works in DNA repair, KDM4A stimulates An unusual and uncontrolled cell proliferation that leads to tumorgenesis cell growth and proliferation Overexpression neoplasic events as well as metastasis or complications related to tumor resilience and spreading speed Lower survival outcomes Inhibiting th pathway and its downstream proteins could help reduce tumor viability, making it easier to deal with it as well as reducing metastasis probabilities Knockdown Higher survival outcomes 11 11 11

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