Biomedical subjects

K Soondrum

Publications and source records attributed to K Soondrum.

3 recordsLinked to original sources

PubMed2001

Human endostatin inhibits growth of human non-small-cell lung cancer in a murine xenotransplant model.

Overall prognosis in human NSCLC remains poor. Antiangiogenic treatment has become a promising concept for the treatment of solid malignancies. Our purpose was to evaluate the efficacy of recombinant HSENDO for the treatment of human NSCLC in an orthotopic murine xenotransplantation model. The efficacy of HSENDO was tested in vitro in cell-proliferation, cell-migration and tube-formation assays. In vivo, the effect of HSENDO on tumor growth was tested in s.c. xenotransplanted human NSCLC and on intrapulmonary induced human NSCLC. In vitro, HSENDO inhibited both human and rodent endothelial cell proliferation in a time- and dose-dependent fashion. Endothelial cell migration was inhibited by 97%. Tube formation of murine endothelial cells was inhibited and preexisting tubes degenerated after HSENDO exposure. In vivo, HSENDO delayed growth of s.c. xenotransplanted tumors. Immunohistochemic staining demonstrated no change in microvessel density but a significant reduction of proliferating tumor cells and an increase in bFGF and VEGF expression, reflecting the antiangiogenic effect of HSENDO. Intrapulmonary tumor induction caused death subsequent to metastatic disease. Systemic HSENDO application extended survival significantly. HSENDO was demonstrated to inhibit endothelial cell proliferation, migration and tube formation effectively. In vivo growth of s.c. transplanted tumors was delayed and survival extended by 32% and 69%, respectively, after intrapulmonary NSCLC induction.

Animals↗

PubMed2001

Detection of micrometastasis through tissue-specific gene expression: its promise and problems.

The detection of micrometastasis holds great promise for earlier staging of patients with malignant diseases and may ultimately guide therapeutic decisions. So far, reverse-transcriptase polymerase chain reaction (RT-PCR) amplification of genes expressed by the tumor in a tissue-specific manner is the method with the highest diagnostic sensitivity. It is well-established that the identification of single tumor cells is feasible in tissues and bodily fluids in both experimental and clinical samples. However, at present it is difficult to assign clinical significance to results obtained from such tests, primarily because their diagnostic specificity is disputed, both conceptionally and methodologically. For example, amplification of candidate mRNA targets is detectable in non-cancer patients using conditions that generally fail to generate such signals from healthy individuals. We have established that transcription of the tissue-specific genes can be affected by different means. Specifically, some target mRNA species are detectable in peripheral blood nuclear cells as low abundance constitutive-like expression, whereas others are induced through in vitro tissue culturing. In addition, mRNA expression may be distinctly upregulated by different cytokines or growth factors in vivo. Also, background transcription of target mRNAs can occur in different lineages of peripheral blood cells. Finally, expression may be substantially different in tissues such as peripheral blood, bone marrow, or lymph nodes. As a consequence, cancer patients in unrelated clinical situations may present with different levels of background expression, making the diagnostic specificity of test results difficult to assess. To add to this complexity, an increasing body of literature is being generated using various targets for a multitude of malignant diseases. There is a great variety of methods for sampling, specimen processing, nucleic acids recovery, test conditions, and readout formats, making it impossible to compare data. In summary, modalities of quantitative RT-PCR methods and standardization issues should be discussed to address these questions.

DNA, Neoplasm↗

PubMed2000

Quantitative PCR.

The classic molecular biology methods like Northern or Southern blot analyse non-amplified DNA or RNA, but need large amounts of nucleic acids, in many instances from tissues or cells that are heterogeneous. In contrast, polymerase chain reaction (PCR)-based techniques allow us to obtain genetic information through the specific amplification of nucleic acid sequences starting with a very low number of target copies. These reactions are characterized by a logarithmic amplification of the target sequences i.e. increase of PCR copies followed by a plateau phase showing a rapid decrease to zero of copy number increment per cycle. Accordingly, the amount of specific DNA product at the end of the PCR run bears no correlation to the number of target copies present in the original specimen. However, many applications in medicine or research require quantification of the number of specific targets in the specimen. This has generated a rapidly increasing need for the development of quantitative PCR techniques. Prominent examples are the determination of viral load in blood specimens for the diagnosis of HIV or HCV infections, the determination of changes in gene dosage through amplification or deletion e.g. of MDR-1, erb-B2, c-myc or the loss of heterozygosity in general. Finally, the analysis of gene expression on the mRNA level does require quantitative approaches to reverse transcriptase PCR, e.g. for studies in morphogenesis or the profiling of cancer cells. Recent advances in technology allow detection of the increment per cycle of a specifically generated PCR product in "real-time mode". Together with the new powerful methods to dissect heterogeneous tissues or fractionate bodily fluids, this now sets the stage for a detailed analysis not only of the genes and genetic changes within a single cell, but also of the use such cell makes of its genes e.g. in pharmacogenomics. Examples of recent developments of the technology and their applications will be given.

Gene Amplification↗