Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Microdissection”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 685 records · Page 38Linked to original sources
PubMed2006

Comparison of gene expression profiles between primary tumor and metastatic lesions in gastric cancer patients using laser microdissection and cDNA microarray.

AIM: To study the differential gene expression profiles of target cells in primary gastric cancer and its metastatic lymph nodes using laser microdissection (LMD) in combination with cDNA microarray. METHODS: Normal gastric tissue samples from 30 healthy individuals, 36 cancer tissue samples from primary gastric carcinoma and lymph node metastasis tissue samples from 58 patients during gastric cancer resection were obtained using LMD in combination with cDNA microarray independently. After P27-based amplification, aRNA from 36 of 58 patients (group 1) with lymph node metastasis and metastatic tissue specimens from the remaining 22 patients (group 2) were applied to cDNA microarray. Semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical assay verified the results of microarray in group 2 and further identified genes differentially expressed in the progression of gastric cancer. RESULTS: The expression of 10 genes was up-regulated while the expression of 15 genes was down-regulated in 22 gastric carcinoma samples compared with that of genes in the normal controls. The results were confirmed at the level of mRNA and protein, and suggested that four genes (OPCML, RNASE1, YES1 and ACK1) could play a key role in the tumorigenesis and metastasis of gastric cancer. The expression pattern of 3 genes (OPCML, RNASE1 and YES1) was similar to tumor suppressor genes. For example, the expression level of these genes was the highest in normal gastric epithelium, which was decreased in primary carcinoma, and further decreased in metastatic lymph nodes. On the contrary, the expression pattern of gene ACK1 was similar to that of oncogene. Four genes were further identified as differentially expressed genes in the majority of the cases in the progression of gastric cancer. CONCLUSION: LMD in combination with cDNA microarray provides a unique support foe the identification of early expression profiles of differential genes and the expression pattern of 3 genes (OPCML, RNASE1 and YES1) associated with the progression of gastric cancer. Further study is needed to reveal the molecular mechanism of lymph node metastasis in patients with gastric cancer.

Aged↗
PubMed2004

Proteomic analysis of human breast cancer tissue with laser-capture microdissection and reverse-phase protein microarrays.

Despite recent advances in breast cancer therapy, women with similar types of breast cancers may respond very differently to standard treatments. The emerging field of clinical proteomics has the potential to revolutionize breast cancer therapy. The ultimate goal of clinical proteomics is to characterize information flow through protein cascades for individual patients. After the protein networks have been elucidated, drug therapies may be specially designed for each patient. The following review describes the proteomic technologies of laser-capture microdissection (LCM) and reverse-phase protein arrays (RPPAs). These technologies allow scientists to analyze relative abundances of key cellular signaling proteins from pure cell populations. Cell survival and apoptotic protein pathways are currently being monitored with LCM and RPPAs at the National Institutes of Health, in phase II clinical trials of metastatic breast and ovarian cancers. Ultimately, proteomics will become an integral component of tracking and managing individualized breast cancer therapy.

Breast↗
PubMed2004

[Application of immunohistochemistry-laser microdissection-PCR technique in detecting p53 gene mutation in paraffin sections of gastric cancer].

OBJECTIVE: To evaluate the application of immunohistochemistry-laser microdissection-polymerase chain reaction (IHC-LMD-PCR) technique in detecting p53 gene mutations using paraffin sections of advanced gastric cancers. METHODS: The expression of p53 protein in 41 paraffin-embedded advanced gastric cancer samples was assessed by immunohistochemistry. The p53-positive or negative carcinoma cells and normal foveolar cells far from the main tumor were then isolated by LMD. The DNA was extracted and p53 gene at exon 5 - 8 amplified by PCR. The products were then analyzed by single-strand conformation polymorphism (SSCP) and automated sequencing. RESULTS: p53 gene was successfully amplified in all the 41 specimens. Overexpression of p53 protein was noted in 11 cases. Mutations of p53 gene were found in 15 cases. Eight of the 11 p53-overexpressed cases had p53 gene mutations. On the other hand, p53 gene mutations were found in only 7 of the 30 p53 protein-negative cases. The presence of p53 gene mutations significantly correlated with p53 protein overexpression (P = 0.004). CONCLUSIONS: IHC-LMD-PCR technique can be successfully applied in paraffin sections of gastric cancers for the detection of p53 gene mutations. The results correlate well with overexpression of p53 protein.

Adenocarcinoma↗
PubMed2004

[The importance and use of laser microdissection in clinical research and diagnostic method development].

The molecular biological analysis of rare cell samples has become increasingly important in medical research and applications. Using laser microdissection and special macromolecule isolation techniques allows us to understand the biological information even in a single cell and to examine the genetical and proteomical features of biologically homogenous cell populations. There are several type of laser microdissectors which have a very user-friendly actuation. In practice the most difficult parts are the sample preparation and the macromolecule extraction from rare cell samples. The aims of this study are to summarize current practical experiences and to foreshow the application possibilities of this quite new technique in clinical practice.

Animals↗
PubMed2005

Global analysis of altered gene expressions during the process of esophageal squamous cell carcinogenesis in the rat: a study combined with a laser microdissection and a cDNA microarray.

The genetic alterations that occur during esophageal tumorigenesis have yet to be determined. We previously established a Wister rat carcinogenesis model of esophageal squamous cell carcinoma. To understand more about the molecular mechanisms during carcinogenesis, we produced esophageal neoplastic lesions by administering N-amyl-N-methylnitrosamine and 12-O-tetradecanoylphorbol-13-acetate to rats. We used laser microdissection to specifically isolate the cells from the normal epithelium, papilloma, dysplasia, and invasive carcinoma. Using a cDNA microarray representing 14,815 clones, we then analyzed the gene expression profiles for each esophageal lesion. The number of differentially expressed genes compared with the normal control dramatically increased in a step-by-step fashion from normal epithelium (1,151 +/- 119 genes) to papilloma (1,899 +/- 543 genes), dysplasia (1,991 +/- 193 genes), and invasive carcinoma (2,756 +/- 87 genes). A hierarchical clustering analysis showed that the three stages of normal epithelium, dysplasia (papilloma), and invasive carcinoma could be clearly classified, whereas the gene expression patterns of papilloma and dysplasia were indistinguishable. Using the Fisher criterion, we also identified 50 genes whose expression level had either significantly increased or decreased in a step-by-step manner from the normal epithelium to dysplasia and then finally to invasive carcinoma. Many of these genes were not previously known to be associated with esophageal carcinogenesis. The present findings in our rat model thus seem to provide us with a better understanding of the molecular alterations that occur during esophageal carcinogenesis and hopefully will also help lead to the development of novel diagnostic and therapeutic targets.

Animals↗
PubMedDate not supplied

Detection of HCV in bile duct epithelium by laser capture microdissection (LCM).

Chronic hepatitis C affects 0.3 to 1.5% of the general population worldwide. The estimated total number of newly acquired hepatitis C virus (HCV) infections is 28,000 in the USA, with 10,000 deaths each year resulting from HCV-associated chronic liver disease. Histological examination of liver tissue from chronic HCV infection shows lymphoid aggregates or follicles in the portal triads, focal fatty change and lobular inflammation. Hepatitis-associated bile duct lesion (HBL) is seen in 5 - 91% of the cases. While the morphological spectrum of HBL has been well described, its pathogenesis in hepatitis C is not known. To this date, evidence supports both the direct injury and immune-mediated mechanisms, but to what extent these mechanisms are involved in the pathogenesis of HBL in chronic hepatitis C remains unclear. Our study showed the presence of HCV in the bile duct epithelium of patients with chronic hepatitis C infection, using the laser capture microdissection technique. These results will enhance our diagnostic capabilities and treatment of chronic hepatitis C infection.

Adult↗
PubMed2006

From tissue phenotype to proteotype: sensitive protein identification in microdissected tumor tissue.

Correlation of disease phenotype with protein profile (proteotype) is a significant challenge for biomedical research. The main obstacles have been the need to insure sufficient quantities of pure protein sample, the reproducibility of protein display, and rapid and accurate protein identification. We present a modified approach that combines enhanced detection sensitivity with tissue microdissection from frozen primary renal cancer tissues of different histological subtypes, followed by 2D gel analysis and protein identification with MALDI mass spectrometry. We obtained reliable and highly consistent results in phenotypically similar tumors of each individual subtype by performing strict morphological control of the analyzed tumor cells without physical or chemical alteration of the frozen tissue samples. By application of non-oxidizing silver staining, proteins were resolved and identified with high levels of specificity and sensitivity. This new combination of techniques allows not only for sensitive identification of specific protein patterns that correspond to a histological tumor phenotype, but also for identification of specific disease-associated protein targets.

Adenoma, Oxyphilic↗
PubMed2006

Intratumoral genetic heterogeneity in pilocytic astrocytomas revealed by CGH-analysis of microdissected tumor cells and FISH on tumor tissue sections.

Pilocytic astrocytomas are the most frequent gliomas of childhood. The majority of cases show cytogenetic normal karyotypes. Although in diffuse gliomas TP53 gene mutations or deletions occur with significant frequency, the role in pilocytic astrocytomas remains unclear. Histomorphologically different areas of 14 pilocytic astrocytomas were microdissected and analyzed for genetic aberrations and heterogeneity. CGH analysis revealed gains of chromosome arm 4q and 6q mainly in areas of classic biphasic pattern, whereas pleomorphic areas presented gains of chromosome 6 and 8q. Using two-color fluorescence in situ hybridization (FISH) the spatial distribution of aneuploidies for chromosomes 7, 8, 17 and the TP53 gene were assessed on parallel sections. FISH analysis revealed a significant percentage of cells with interspersed heterozygous deletions of TP53 in all tumors (14/14), ten cases showed also monosomy 17. Besides gains of chromosomes 7 and 8, losses of these chromosomes were detected in the majority of tumors. In conclusion, pilocytic astrocytomas show a genetic heterogeneity associated with variations of histologic structure as well as an intratumoral heterogeneity observed on single cell level by FISH.

Adolescent↗
PubMed2006

Loss of imprinting in IGF2 in colorectal carcinoma assessed by microdissection.

To ascertain the implications of loss of imprinting (LOI) of the insulin-like growth factor II gene (IGF2) for carcinogenesis, the precise frequency of LOI in colorectal carcinoma was examined using a laser capture microdissection method, and compared to the matched normal colorectal mucosa. LOI was examined by PCR-restriction fragment length polymorphism in combination with direct sequencing. The status was assigned as imprinting when PCR-RFLP showed only one band or sequence with a single peak, otherwise cases were classified as LOI. LOI was found in 13 of 24 informative cases of carcinoma (54%), which was higher than the ratios reported previously. LOI was also found in the normal colorectal mucosae in 14 cases (58%). The LOIs in carcinomas and in the normal mucosae were closely correlated: 10 of 13 LOI-positive carcinomas showed LOI in the matched normal mucosae. These results suggest that LOI of IGF2 in colorectal carcinoma and LOI in the background mucosa play important roles in carcinogenesis.

Binding Sites↗
PubMed2006

[An application of laser capture microdissection to isolate the pure atrial cells].

OBJECTIVE: To use the laser capture microdissection (LCM) to isolate the pure atrial cells from heart. To extract tiny amount RNA and amplify the genes of beta-actin and GAPDH. METHODS: First, the prepared sample RNA was extracted and verified. Second, the frozen sections were scraped after haematoxylin-eosin staining, and then the RNA of scraped tissues was extracted and verified. Last, the tiny amount RNA was extracted from atrial cells captured by LCM, and the genes of beta-actin and GAPDH were amplified by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: The atrial cells isolated with LCM had clear morphology after quick staining. The high quality RNA was extracted from LCM sample and scraped tissues, of which A260/A280 was 1.9-2.0. And 18S rRNA and 28S rRNA were seen distinctly on the gel of RNA formaldehyde denaturing gel electrophoresis, which indicated that RNA didn't degrade before cells captured. Too small amount of RNA extracted from captured cells was below the detectable limit of gel electrophoresis or ultraviolet spectrophotometer. Bands of 300 bp and 357 bp could be amplified by RT-PCR from tiny amount RNA. CONCLUSION: Highly pure atrial cells could be obtained by LCM. Tiny amount RNA of captured atrial cells could be extracted and amplified with genes of beta-actin and GAPDH.

Actins↗
PubMed2006

[Laser capture microdissection and its application in andrology].

Laser capture microdissection (LCM) is a novel technique developed in the recent years. Under direct microscopic visualization, LCM permits rapid one-step procurement of the selected pure target cell subgroup, even a single cell from a section of the complex, heterogeneous tissue. This technique has rapidly been introduced in the researches for the cellular and molecular mechanisms of diseases. It helps improve diagnosis, therapy and prognosis. This article reviews the progress in the application of LCM in andrology.

Andrology↗
PubMed2006

Blunt microdissection and rotational atherectomy: an effective combination for the resistant chronic total occlusion.

Percutaneous coronary intervention (PCI) of chronic total occlusions (CTO) has a lower success rate than nontotal occlusions. Like nontotal coronary occlusions, successful procedural outcomes of CTOs confers improvement in clinical outcomes. Reasons for unsuccessful PCI of CTOs may stem from various steps of the procedure including failure to cross the occlusion with a guidewire or a balloon, or failure to adequately dilate or modify the lesion. The Frontrunner blunt microdissection device is designed to improve the odds of guidewire passage through coronary occlusions, whereas the Rotablator rotational atherectomy device is intended for plaque modification, especially in resistant, elastic or calcific lesions. The Frontrunner and Rotablator can each facilitate the procedure at their respective junctures; the combined use of these technologies can enhance the overall PCI success rates, especially regarding difficult CTOs. This case presentation encounters two technical challenges: the inability of the balloon catheter to cross the CTO, and the inability to recross the occlusion with the less favorable RotaWire. In this case, the combined use of the Frontrunner and Rotablator devices was necessary to achieve overall procedural success. When tackling difficult CTOs, a comprehensive strategy that encompasses multiple devices and technologies can enable ultimate procedural success.

Aged↗
PubMed2006

Expression status of RUNX1/AML1 in normal gastric epithelium and its mutational analysis in microdissected gastric cancer cells.

Although Runt-related transcription factors RUNXs (RUNX1-3) have a high similarity in their structure, only RUNX3 is known to be involved in gastric carcinogenesis. First, we examined mRNA expression of these three RUNX genes in the gastric mucosa, and, finding only RUNX2 was not expressed there, we further investigated RUNX1 and RUNX3 expression in three regions including the pit, isthmus/neck, and gland regions of the human normal stomach and whether RUNX1 is involved in gastric carcinogenesis. The mRNA expression of RUNX1 and RUNX3 was examined by use of the three regions isolated by laser-captured microdissection (LCM) and by use of primary gastric cancer tissues. Furthermore, RUNX1 mutational analysis was performed in the cancer cells, which also were isolated from 44 paraffin-embedded gastric cancer tissues by LCM. RUNX1 was co-expressed with RUNX3 in the pit region, and has cell growth-inhibition activity similar to RUNX3. RUNX3 has been reported to be suppressed by DNA methylation in a subset of gastric cancers; however, the expression of RUNX1 mRNA was observed in all of the gastric cancer cell lines and gastric cancer tissues that we examined. No RUNX1 mutation was found in the 44 gastric cancer patients. Although RUNX1 is similar to RUNX3 in both the expression pattern in the stomach and its cell growth-inhibition activity, RUNX1 is not involved in most cases of gastric cancers. These results suggest that the transcriptional target genes are different between these two family genes.

Cell Differentiation↗
PubMed2005

Clinical and microdissection genotyping analyses of the effect of intra-arterial cytoreductive chemotherapy in the treatment of lacrimal gland adenoid cystic carcinoma.

PURPOSE: To determine the effect of intra-arterial cytoreductive chemotherapy (IACC) as an adjunct of a multimodality protocol for the treatment of lacrimal gland adenoid cystic carcinoma (ACC). METHODS: This was a retrospective, comparative, consecutive case series. Nine consecutive patients with lacrimal gland ACC were treated with IACC, followed by orbital exenteration and chemoradiotherapy. This case series was compared with a series of seven patients treated by conventional local therapies. Clinical records, imaging studies, histologic sections, and archival specimens from all 16 patients were reviewed. Information analyzed included site of disease, histologic characteristics, extent of disease, local-regional recurrence or distant metastases, and disease-free survival time. Gene analysis was performed on microdissected tissue samples. Mutational allelotyping targeting nine genomic loci using 15 polymorphic microsatellite markers situated in proximity to known tumor suppressor genes serve as markers for the presence of gene deletion. The effect of IACC was assessed by the radiographic response and survival outcome in comparison to a historical cohort of patients managed by conventional local therapies. A fractional mutation index was used to compare the acquired mutational load between different tumors having nonidentical patterns of microsatellite informativeness. RESULTS: The carcinoma cause-specific death rates between the two treatment groups was significant (P = .029, log-rank test). The cumulative 5-year carcinoma cause-specific death rate was 16.7% in the IACC-treated group compared with 57.1% in the conventional treatment group. 1p36 was the single most common site affected by allelic loss for microsatellite markers in this series. CONCLUSIONS: The preliminary data suggest that IACC as an integral component of a multimodal treatment strategy is potentially effective in improving local disease control and overall disease-free survival in lacrimal gland ACC. Allelic loss for microsatellite markers at 1p36 may be a common and an early event in ACC formation and progression.

Adult↗
PubMed2006

Gene expression profiling of lymph node metastasis by oligomicroarray analysis using laser microdissection in esophageal squamous cell carcinoma.

We applied oligomicroarray analysis of 17086 genes to identify the genes related to lymph node metastasis in esophageal squamous cell carcinoma (ESCC). The samples of cancer and non-cancerous paired tissue were taken from 16 patients with ESCC who underwent esophagectomy with lymph node dissection. Total ribonucleic acid was extracted from the cancer cells obtained by using laser microdissection and was amplified by T7 based-amplification for the application to the oligomicroarray. The oligomicroarray demonstrated 43 overexpressed genes, such as cell-cycle regulators, cell adhesion related genes, anti-apoptosis related genes, and 138 suppressed genes such as cell differentiation related and apoptosis related genes in ESCC cells with lymph node metastasis. Among them, 5 overexpressed genes (SPP-1, CKS2, CCT5, STMN1, NDUFB9) and one suppressed expression gene (GJB2) were selected in the gene profiles, and then the expressions of those genes were confirmed by real-time semi-quantitative reverse transcriptional polymerase chain reaction (RT-PCR) method for confirmation of the result not only in study cases but also in additional 21 cases. The gene expression by real-time semi-quantitative RT-PCR was in accordance with the microarray data. Although we were able to extract some genes related to nodal metastasis in ESCC, further examination is necessary in other genes as well as the interaction of stromal tissues.

Aged↗
PubMed1990

Vascular microdissection in strabismus surgery.

Microdissection of the muscular vessels allows recessions, resections, or transpositions of the recti muscles without interrupting the blood flow to the corresponding anterior ciliary arteries. This avoids postoperative impaired blood flow to the anterior segment and may play a role in preventing anterior segment ischemia syndrome. The use of a fine-hooked cannula and microscissors, in addition to the usual strabismus surgery instruments, is described.

Dissection↗
PubMed1999

Loss of heterozygosity, differentiation, and clonality in microdissected male germ cell tumours.

Testicular germ cell tumours (TGCTs) are heterogeneous neoplasms with different histological patterns and malignant potential. The aim of this study was to determine whether the main TGCT subtypes (seminoma, embryonal carcinoma, yolk sac tumour, choriocarcinoma, and mature teratoma) are distinguished by their loss of heterozygosity (LOH) patterns and whether LOH typing can help to distinguish between clonal and multifocal development of different components in mixed TGCTs. In 76 tumours analysed for allelic losses at 25 chromosomal loci, different LOH patterns were found in distinct histological subtypes. A region around D18S543 frequently lost in yolk sac tumours could harbour one or more tumour suppressor genes. In 20 microdissected mixed tumours, losses of identical alleles in different histological components in 11 of 20 cases (over 50 per cent) were found, which is in favour of current histogenetic models of clonal TGCT development. Clonal losses were most often found at D13S317 (6 of 20 tumours). Two classes of allelic losses may therefore occur during TGCT development: clonal losses which are involved in early transformational events and others related to TGCT differentiation along different lines.

Adult↗
PubMed2000

Comparative genomic hybridization of microdissected samples from different stages in the development of a seminoma and a non-seminoma.

Human testicular germ cell tumours (TGCTs) of adolescents and adults, both seminomas and non-seminomas, originate from intratubular germ cell neoplasia (IGCN). Comparative genomic hybridization (CGH) was applied to microdissected samples from different stages of the development of a seminoma and a mixed non-seminoma, including IGCN of both. The different stages of the seminoma development, namely IGCN, intratubular and invasive seminoma, showed a very similar pattern of chromosomal imbalances, including gains of parts of 7, 8, 12,14, and X, and losses of parts of 3, 4, 5, 10, 11, 12q, 16, 18, 22, and Y. A more heterogeneous pattern was found for the non-seminoma. Some aberrations were present only in IGCN, or in IGCN and in all invasive components (gains of parts of 1q, 17, 19p, 20q, and 22, and losses of parts of 4, 5, 9p, 13, and 18q), while others were present in a less consistent pattern. These are the first reported CGH data from different stages in the development of TGCTs. Although only two cases were studied, the results suggest that particular numerical changes of (parts of) chromosomes are involved in the early development and progression of this cancer.

Adolescent↗