Mereja

Author: Kok Siong Chen

  • Cytogenetic Studies in Malignancy Cancer

    I think I never mention about the relationship between cytogenetics and cancer research in this blog. Cytogenetic studies are proven that can assist to define the diagnosis and deliver clear prognostic differences in cancer. There are many cytogenetic studies of malignancy have been gone through and most of them are related to hematologic disorders. However, those studies constitute only small percentage of all cancer malignancies.
     

    Clinical Value of Cytogenetic Studies in Malignancy Cancer

    cytogenetics cancer Cytogenetic Studies in Malignancy Cancer

    Recently, there has been a significant progress in knowledge of the recurrent abnormalities in many of solid tumors as well as hematologic malignancies. Consequently, the clinical value of cytogenetic studies in cancer malignancy has been appreciated after the cytogenetic studies of some solid tumors moving out of cancer research environment.
     
    Apart from this, there would be little clinical value in doing cytogenetic studies in cancer malignancy if all the patients with a particular cancer died. The researchers only manage to probe the origins of cancer for those patients. However, after researching in all tumors type by using cytogenetic technique, the presence or absence of many of the genetic abnormalities found has been related to different responses to treatment. Since that, the genetic and cytogenetic studies start to be recognized as important and one of the best choices of treatment for a cancer patient.
     

    Cytogenetic Studies Helps Diagnostic Investigation in Malignancy Cancer

    Consequently, cytogenetic analysis of cancer malignancy is considered to provide rapid, accurate and specific results to aid the clinical professionals. This can help to choose the most appropriate type of treatment to the cancer patient in shorter time. However, John Swansbury stated that the diagnosis of a malignancy can be traumatic, and an accurate and early indication of every patient’s prognosis is valuable.
     
    In conclusion, a cytogenetic study remains an essential part of diagnostic investigations of every patient with hematologic malignancy and certain solid tumors without denying the valuable contributions made by other genetic assays.

    Reference: Methods in Molecular Biology, vol. 220: Cancer Cytogenetics: Methods and Protocols. Edited by: John Swansbury © Humana Press Inc., Totowa, NJ
     

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    Cytogenetic Studies in Malignancy Cancer is a post from: Cytogenetics and Cancer Research

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  • Protein Extraction for Western Blotting in Cancer Research

    Protein extraction is an important step for Western Blotting in cancer research. The proteins from samples (cancer cell line) need to be extracted efficiently without degradation. I have learnt this technique during laboratory work for my cancer research project. Although the protocol of this protein extraction technique is quite scientific and technical, I still wish to share as maybe someone who is going to do the cancer research will find it useful. I’m going to explain the nuclear and cytoplasmic protein extraction technique.
     

    3 Reagents in Protein Extraction

    protein extraction western blotting cancer research

    Before we start to do the protein extraction, we need to prepare the reagents and the equipments. There are 3 reagents (CER I, CER II and NER) that we need in nuclear and cytoplasmic protein extraction.  Before the reagents can be used, we have to add protease and phosphatase inhibitor cocktails to these 3 reagents. The protease and phosphatase inhibitor cocktails provide the solution with full sample protection to protect the protein from degradation during the extraction. In my cancer research, I add 1:100 ratios of protease and phosphatase inhibitor cocktails to the reagents. As I need 200 µl of CER I for each sample to be extracted, I have to add 2 µl of protease and phosphatase inhibitor cocktails to mix with the reagent.
     

    Protocol of Protein Extraction

    Now we can start the protocol of nuclear and cytoplasmic protein extraction.
     
    1. Firstly, we have to spin down 20 µl of cells into a 1.5 ml microcentrifuge tube at 500 xg for 3 minutes. After spinning, the supernatant was discarded using a pipette.
     
    2. Then, we have to add 200 µl of ice cold CER I to the cell pellet. The mixture is vortex vigorously for 15 seconds so that the pellet is fully resuspended. The tube is incubated on ice for 10 minutes.
     
    3. Consequently, we have to add 11 µl of ice cold CER II and vortex again for 5 seconds. Then, the tube is incubated on ice again for 1 minutes and vortex for 5 seconds too.
     
    4. Next, we need to centrifuge the mixture. Centrifugation is the process to separate the contents in the mixture. After the centrifugation at 16,000 xg for 5 minutes, we need to transfer the supernatant which is the cytoplasmic extract to a new pre-chilled tube. Just keep the tube on ice until used.
     
    5. Then, we need to extract the nuclear protein. The insoluble pellet is resuspended with 100 µl of ice cold NER and vortex for 15 seconds.
     
    6. The sample is put on the ice for 10 minutes and vortex for 15 seconds. This step is repeated 4 times for a total of 40 minutes.
     
    7. Later, the sample is centrifuged again at 16,000 xg for 10 minutes. The supernatant which is nuclear extract is transferred to a clean pre-chilled tube and keep it on ice until used. The nuclear and cytoplasmic protein extraction for Western Blotting in cancer research is done.
     
    After the protein extraction, we need to continue with the quantification of protein concentration by using Bradford assay before we can proceed to the SDS-PAGE. I will learn how to quantify the protein concentration myself next time.

    Adapted from: NE-PER Nuclear and Cytoplasmic Extraction Kit (C/N: 78833)
     

    Do you ever heard about protein extraction? Have you do it before? Come and share with us.

    Protein Extraction for Western Blotting in Cancer Research is a post from: Cytogenetics and Cancer Research

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  • Cancer Research on Natural Compound (Cancer Review)

    I have started my final year project about cancer research today. My project title is “Natural Compound: Modulating the Expression of Both Anti- and Pro-apoptotic Proteins for Cancer Therapy”. Initially, i need to write a literature review on this cancer research project so that i can fully understand what i’m going to do. Today, i wish to post up my simple review on cancer.
     

    Cancer Review

    cancer research on natural compound
    Cancer is a group of diseases that are characterized by uncontrolled cellular growth, enhanced angiogenesis and reduced programmed cell death (David and Lisa, 2008). Cancer can spread to other locations throughout the body, where they grow and upset normal bodily functions. We called this as metastatic spread where the cells are dispersed around the blood stream that formed leukemia or the lymphatic system that formed lymphoma. On the other hand, some of the cancer cells will clump together to form solid tumors (Arthur and Gary, 2009). The disruptive behaviours of cancer cells are because of the changes in their genomes and gene expression that result in disruption of normal regulatory signalling pathways (Atif and Peter, 2006).
     

    Carcinogenesis

    Carcinogenesis is the process of which normal cells are transformed into cancer cells (Atif and Peter, 2006). About 200 varieties of cancer disease have been described. The cancer diseases can be grouped in three main types: carcinomas, sarcomas, leukemias and lymphomas. Carcinomas are solid cancers arisen from the epithelial cells that covering our inner and outer surfaces. Sarcomas are solid cancers that developed from the connective tissue cells that form our body structures. Leukemias and lymphomas are cancers that caused by white blood cells (Arthur and Gary, 2009).
     

    Tumorigenesis

    Tumors are neoplasm which a population of genetically related cells has acquired the ability to proliferate abnormally. The malignant tumors have the ability to invade surrounding tissues composed of normal cells and make them lethal. Tumorigenesis is the process of the growth of a tumor from a single genetically altered cell (Fred, 2008).
     

    Cancer Risk Factors

    There are many risk factors of cancer which include both external factors and internal factors. Some of the examples of external factors are tobacco use, chemical use, radiation, and infectious organisms that will induce the developing of cancer cell. On the other hand, the internal factors such as inherited mutations, hormones, immune conditions, and mutations that involved the metabolism are the risk factors that causing the developing of cancer cell. These causal factors may act together or in sequence to initiate or promote carcinogenesis (American Cancer Society, 2008).
     
    That’s all for my simple cancer review. I will continue to write more reviews later on. Stay tuned!:)
    References:
    1. David S. A. & Lisa M. H. (2008). Fundamental of Cancer Prevention (Second Edition). German: Springer-Verlag Berlin Heidelberg.
    2. Arthur B. P. & Gary S. S. (2009). The Biology and Treatment of Cancer. Canada: John Wiley & Sons, Inc.
    3. Atif B. A. & Peter G. B. (2006). Nutrition and Cancer Prevention. Boca Raton: Taylor & Francis Group.
    4. Fred B. (2008). Principle of Cancer Genetics. Maryland: Springer Science + Business Media B. V.
    5. American Cancer Society. Cancer Facts and Figures 2008. American Cancer Society, Atlanta, GA, 2008.
     

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    Cancer Research on Natural Compound (Cancer Review) is a post from: Cytogenetics and Cancer Research

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