Antigen Messenger Ribonucleic Acid Research Articles

Detection of CEA mRNA on non-small lung cancer and it's significance.

In recent years, many studies on micrometastasis in non-small cell lung cancer (NSCLC) have been reported, this study is to investigate the effect of operation on micrometastasis from NSCLC and evaluate the relation between micrometastasis and clincopathological parameters. The blood samples were taken from 70 cases of NSCLC and 18 patients with benign diseases at 3 intervals during the operation from peripheral vein. The transcription of carcinoembryonic antigen messenger ribonucleic acid (CEA mRNA) was assayed by means of nested reverse transcriptase polymerase chain reaction (RT-PCR) and micro-fluid chip. The CEA mRNA positive rates of all 3 time spots were as follows: 50% at beginning of the operation (Time 1), 62.8% at ligating the pulmonary vein (Time 2) and 57.1% at 1 h after ligating pulmonary vein (Time 3). There is significant difference between Time 1 and Time 2 (Chi-Square=7.114, P <0.05). The positive rates of well-differentiation and middle-differentiation, stage I and state II, Tis, T1 and T2, N0 were significant less than non-differentiation and low-differentiation, stage III and state IV, T3 and T4, N1, N2 and N3, respectively. No negative control samples was found to be positive, and no positive control samples was found to be negative. The sensitivity of our test was 10 cells/mL. The cancer cells dissemination during operation was demonstrated indirectly in our study, the time of pulmonary vein ligation (earlier or later) may affect the quantity of tumor cells released into circulation; The patients with lower differentiation, advanced TNM stage, larger tumor size and metastasis of lymph node have higher rates of metastasis in peripheral, so the detection of CEA mRNA can guide the therapy of NSCLC to a certain extent.

Molecular profiling of polycystic ovaries for markers of cell invasion and matrix turnover

To study gene expression profiles of connective tissue components in polycystic ovaries using complementary deoxyribonucleic acid (cDNA) array technology. Descriptive study of normal and polycystic human ovarian biopsy samples analyzed by cDNA array hybridizations. Experimental laboratory research. Eight women with polycystic ovary syndrome (PCOS) and two normally cycling women treated with electrocauterization and hysterectomy, respectively. Ovarian biopsy samples. Expression levels of 588 genes involved in cellular invasion, extracellular matrix (ECM) turnover, and cell-ECM interactions in polycystic ovaries. A majority of the 30 genes down-regulated in PCOS ovaries represented those related to cell adhesion and motility, as well as angiogenesis, followed by regulators of cell cycle and growth. The 14 up-regulated genes represented those regulating cell fate and development, growth factors, cytokines, chemokines, and cell-cell interactions. Of the 44 transcripts exhibiting marked changes in the cDNA array analysis, only one - proliferating cell nuclear antigen messenger ribonucleic acid (PCNA mRNA) - was systematically down-regulated; 2 transcripts, for CDC27HS protein and CD9 antigen, were down-regulated in 7 out of 8 PCOS samples. The present data suggest that gene expression profiling may become a useful tool to classify PCOS patients into subgroups with different etiologies. Genome-wide expression profiling using microarrays should be performed to better understand the metabolic derangement(s) in PCOS.

A case of leptomeningeal carcinomatosis from esophageal basaloid carcinoma diagnosed by quantitative reverse transcription-polymerase chain reaction for carcinoembryonic antigen

We describe herein the case of a 68-year-old man who experienced leptomeningitis after esophagectomy from an esophageal basaloid carcinoma. Although the patient had a good operative course for the first 10 days after surgery, he suddenly had general convulsions with unconsciousness. He was placed on mechanical ventilation in an intensive care unit. Computed tomography and magnetic imaging resonance did not reveal any abnormal findings. No abnormal data in the cerebrospinal fluid were found by biochemical, virological, and cytological examination. The positive expression of carcinoembryonic antigen messenger ribonucleic acid in cerebrospinal fluid was detected by a quantitative reverse transcription-polymerase chain reaction method. Immunohistochemical staining using an anticytokeratin antibody confirmed the presence of tumor cells in the cerebrospinal fluid. Spontaneous breathing was recovered after treatment with systemic chemotherapy. Six months after surgery, computed tomography revealed multiple brain metastases. This case demonstrates that the quantitative reverse transcription-polymerase chain reaction method of analyzing carcinoembryonic antigen messenger ribonucleic acid may be a sensitive and useful method for determining leptomeningeal metastasis before the detection of tumors by cytological and imaging examinations in patients with cancer.

Carcinoembryonic antigen mRNA analysis detects micrometastatic cells in blood from lung cancer patients.

The current authors previously identified circulating cells expressing carcinoembryonic antigen (CEA) messenger ribonucleic acid (mRNA) in 80% of lung cancer patients bearing distant metastases. The current study prospectively validated the data on a novel cohort and extended the study to other mRNAs expressed by neoplastic cells. CEA, cytokeratin 19 and 20, aldolase A and epithelial glycoprotein 2 (EPG2) mRNA was analysed by reverse transcriptase-polymerase chain reaction in circulating cells from 19 healthy controls, and in biopsies and blood at diagnosis from 32 lung cancer patients monitored for 24 months. Aldolase A and cytokeratin 19 mRNA occurred in circulating cells of all controls; cytokeratin 20 was not expressed by any lung cancer biopsy. EPG2 mRNA occurred in all biopsies but not in the patients' circulating cells. CEA mRNA occurred in 29/32 (90.6%) biopsies and in 17/32 mRNA samples from circulating cells from lung cancer patients. Of these positive patients 12/17 developed metastases within 9 months of mRNA analysis. Three positive patients died, one was lost to follow-up, and one did not develop metastases within 24 months. Of the negative patients 12/15 did not develop metastases during the 24-month follow-up; one patient was lost to follow-up, one did not express CEA, and another developed metastases. Unlike in other neoplasias, cytokeratin 19 and 20, aldolase A and epithelial glycoprotein 2 messenger ribonucleic acid are not useful for the detection of circulating cancer cells in lung cancer. Carcinoembryonic antigen messenger ribonucleic acid analysis in circulating cells helps to identify lung cancer patients at a greater risk of metastases.

Clinical impact of carcinoembryonic antigen messenger ribonucleic acid expression in tumor-draining vein blood on postoperative liver metastasis in patients with colorectal carcinoma: a prospective, cohort study.

This study was designed to determine the clinical impact of carcinoembryonic antigen messenger ribonucleic acid expression in tumor-draining vein blood on postoperative liver metastasis in patients with colorectal carcinoma. The study comprised 37 patients with colorectal carcinoma who underwent laparotomy (Dukes A, 3; Dukes B, 16; Dukes C, 15; and Dukes D, 3). Blood samples were drawn from the tumor-draining vein and the tumor-feeding artery. Total ribonucleic acid was extracted from the blood and subjected to reverse transcriptase-polymerase chain reaction to detect carcinoembryonic antigen messenger ribonucleic acid. Tissue sections of tumor were stained with hematoxylin and eosin and were examined immunohistochemically. The duration of follow-up ranged from two years and three months to three years and six months. Expression of amplified carcinoembryonic antigen messenger ribonucleic acid in tumor-draining vein blood was positive in 16 (43 percent) of the 37 patients, while that in the feeding artery blood was positive in 3 patients (8 percent). There was no relation between the protein level of carcinoembryonic antigen and positive expression of carcinoembryonic antigen messenger ribonucleic acid. Positive expression of carcinoembryonic antigen messenger ribonucleic acid was observed in two patients with histologic evidence of massive venous invasion and in all three patients with synchronous liver metastasis. Immunohistochemical studies revealed that all tumor specimens with stromal type (5/5) of carcinoembryonic antigen staining had positive expression of carcinoembryonic antigen messenger ribonucleic acid. Postoperative liver metastasis was detected in four patients, three of whom (75 percent) had positive expression of carcinoembryonic antigen messenger ribonucleic acid in tumor-draining vein blood. Positive expression of carcinoembryonic antigen messenger ribonucleic acid in tumor-draining vein blood differed by the type of immunohistochemical staining with carcinoembryonic antigen. These data suggest that patients with positive expression of carcinoembryonic antigen messenger ribonucleic acid in tumor-draining vein blood may be at a high risk for postoperative liver metastasis.

Detection and diagnostic significance of carcinoembryonic antigen messenger ribonucleic acid in lung cancer tissues

To evaluate the clinical value of carcinoembryonic antigen messenger ribonucleic acid ( CEA-mRNA) detection in the diagnosis of lung carcinoma. CEA-mRNA expression was detected in 32 patients with lung carcinoma by reverse transcriptase-polymerase chain reaction ( RT-PCR) , and 11 patients with benign pulmonary diseases as control. The positive rate of CEA-mRNA was 84. 4% ( 27/ 32) in lung cancer group. And there was no positive case of CEA-mRNA in control group. The specificity was 100% and the sensitivity was 84. 4% in the diagnosis of lung carcinoma. CEA-mRNA detection by RT-PCR method may be helpful to diagnosis and differential diagnosis of lung cancer and be of clinical value.

MOLECULAR DETERMINATION OF SURGICAL MARGINS USING FOSSA BIOPSIES AT RADICAL PROSTATECTOMY

Status of the surgical margins after radical prostatectomy is a key factor for predicting postoperative outcome. Current methods used to determine margin status are tedious, costly and vary among institutions. Sensitive and inexpensive detection of prostate cells in the circulation of patients with prostate cancer has been achieved using reverse transcriptase (RT) polymerase chain reaction (PCR) for prostate specific antigen and prostate specific membrane antigen. Therefore, we designed and tested a novel and objective molecular assay for assessing surgical margins at radical prostatectomy based on the detection of prostate specific markers using RT-PCR. We also compared this assay to standard pathological examination. A total of 30 consecutive patients with local prostate cancer underwent radical prostatectomy. At the completion of gland excision 5 biopsies of the prostatic fossa were obtained for histopathological and molecular analysis. We performed RT-PCR analysis for prostate specific antigen and prostate specific membrane antigen messenger ribonucleic acid in these biopsy specimens, and compared the results with pathological stage. Men free of prostate cancer who underwent radical cystoprostatectomy for bladder cancer or abdominoperineal resection for rectal cancer served as controls. There were positive molecular margins in all patients with positive margins and/or extracapsular extension. No controls had a positive molecular assay. In 4 of 16 patients (25%) histopathological evaluation revealed organ confined disease but biopsies were positive by the molecular assay, including those in 2 (50%) who had been treated with neoadjuvant hormonal therapy before surgery because of a higher estimated risk of extracapsular disease. Results in 4 cases were uninformative. Our results with an objective molecular assay aimed at assessing surgical margins after radical prostatectomy reveal an excellent correlation with conventional pathological analysis. In addition, molecular assessment of the prostatic fossa identifies patients in whom extracapsular disease may have been unidentified by conventional pathological examination. In addition, this assay yields clues to why neoadjuvant hormonal treatment before radical prostatectomy does not seem to decrease the biochemical failure rate in these patients. Larger studies with longer followup are required to determine the prognostic significance of these positive molecular margins.

Enzyme-linked immunosorbent assay detection of prostate-specific antigen messenger ribonucleic acid in prostate cancer

Objectives. To develop a rapid, sensitive, reverse transcriptase-polymerase chain reaction (RT-PCR) prostate-specific antigen (PSA) messenger ribonucleic acid (mRNA) detection method by applying colorimetric enzyme-linked immunosorbent assay (ELISA). Methods. Total RNA was extracted from 16 urogenital cancer cells (including PSA-producing LNCaP cells) from pelvic and inguinal lymph node aspiration biopsy samples from patients with prostate, bladder, and penile cancer, as well as from blood samples of 500 patients with urogenital cancer. We used r Tth polymerase for RT and PCR. The RNA target was amplified by RT-PCR with dinitrophenyl-labeled primer. The PCR product was denatured and hybridized on a PSA-specific probe-coated microwell plate. Results. In 16 cancer cell lines, only LNCaP cells expressed especially high PSA mRNA values, with an optical density (OD) greater than 3. In other cell lines, two testicular cells had relatively high ODs, 1.909 and 0.987, respectively. A high PSA mRNA value was obtained by fine needle aspiration from pelvic lymph node specimens of cytologically positive lymph nodes from patients with prostate cancer but not from patients with cytologically proved bladder or penile cancer. Sensitivity and specificity of fine needle aspiration samples were 70% and 100%, respectively. Blood tests obtained from patients with prostate cancer demonstrated high PSA mRNA values. Conclusions. The PSA mRNA RT-PCR ELISA method provides a sensitive photometric enzyme immunoassay for the detection of PSA mRNA, using nonradioactive techniques.

Detection of micrometastases in histologically negative lymph nodes in esophageal cancer

Background. New molecular techniques may identify micrometastases in histologically negative lymph nodes and have an impact on the staging of esophageal cancer. We investigated the role of the reverse transcriptase–polymerase chain reaction (RT–PCR) assay to identify micrometastases in esophageal cancer. Methods. The RT–PCR assay to detect carcinoembryonic antigen (CEA) messenger ribonucleic acid (mRNA) was performed on lymph nodes from patients with esophageal cancer and benign esophageal disorders. The presence of CEA mRNA in lymph nodes was considered evidence of metastases. Results. Histopathologic study revealed metastases in 50 (41%) of 123 lymph nodes from 30 patients with esophageal cancer. All histologically positive lymph nodes contained CEA mRNA by RT–PCR. Of 73 histologically negative lymph nodes, 36 (49%) contained CEA mRNA, a significant increase compared with the histopathologic diagnosis ( p < 0.001). Lymph nodes in patients with benign disease contained no CEA mRNA. In 10 patients, histologic stage was N0. Five of them were also negative by RT–PCR, and all are alive with only one recurrence. In the remaining 5 patients, RT–PCR was positive for occult lymph node metastases; 2 have died of disease, and 1 is alive with recurrent disease. Conclusions. In patients with esophageal cancer, RT–PCR detects more lymph node metastases than does histopathology. Initial follow-up suggests a positive RT–PCR with negative histologic findings may have poor prognostic implications. Further studies will be needed to confirm any clinical implications.

The sequence of vessel ligation affects tumor release into the circulation

Objective: Whether the sequence of pulmonary vein and artery ligation in pulmonary lobectomy for carcinoma affects intraoperative hematogenous cancer cell dissemination is not known. We examined whether vessel ligation sequence affects the presence of circulating cancer cells as reflected by carcinoembryonic antigen messenger ribonucleic acid. Methods: We assayed for the transcripts of carcinoembryonic antigen messenger ribonucleic acid by reverse-transcriptase polymerase chain reaction in peripheral blood taken before, during, and after operation from 30 patients with non–small-cell lung cancer who underwent a curative lobectomy and from six patients with limited-stage small-cell lung cancer who were treated initially with chemotherapy followed by lobectomy. Each patient was randomly assigned before the operation to have either pulmonary vein ligation or pulmonary artery ligation first. Blood taken from 10 patients with interstitial pulmonary fibrosis who underwent an open lung biopsy and 41 healthy subjects served as a control. Results: No control samples were positive for transcripts. Sixteen of the preoperative blood samples from the 30 patients with non–small-cell cancers were positive. Of these 16, eight samples remained positive even after lobectomy was performed; the remaining eight samples (four in each ligation group) became negative. Of the 14 initially negative samples (seven in each ligation group), nine samples became positive during the operation. Such conversion during the operation was more common with arterial ligation first (six patients, 85.7%) than with venous ligation first (three patients, 42.9%). Samples from all six patients with small-cell cancer were positive before the operation, and five of six samples remained positive after the operation. Conclusions: Many patients with non–small-cell lung cancer have systemic disease even when they were thought to have resectable tumors. Ligating the pulmonary vein before ligating the artery may lessen intraoperative hematogenous dissemination. Most small-cell lung cancers represent systemic disease even when considered resectable. (J Thorac Cardiovasc Surg 1998;116:107-13)

Cryopreserved aortic homografts contain viable smooth muscle cells capable of expressing transplantation antigens

Frozen aortic tissue is increasingly used as homografts in reconstructive cardiovascular surgical procedures. The viability of cells within these cryopreserved tissues, their identity, and their potential immunogenicity have been the subject of controversy. We cultured cells from cryopreserved human aortic homografts that reacted with a monoclonal antibody that recognizes muscle actin isoforms, identifying them as smooth muscle cells. Under basal conditions, these smooth muscle cells contained messenger ribonucleic acid for class I human leukocyte antigens detected by northern blotting and expressed class I human leukocyte antigen on their surfaces as measured by enzyme-linked immunoassay and immunohistochemistry. Unstimulated smooth muscle cells contained no class II human leukocyte antigen messenger ribonucleic acid as determined by northern blotting and displayed almost no class II surface antigen as determined by enzyme-linked immunoassay. Interferon gamma (1000 U/ml, 72 hours), a product of activated T lymphocytes, not only increased the expression of class I human leukocyte antigens by smooth muscle cells, but induced class II human leukocyte antigen messenger ribonucleic acid and elevated surface expression from 22 +/- 7 to 819 +/- 35 enzyme-linked immunoassay units (n = 4). Immunohistochemistry revealed few class II-positive smooth muscle cells under basal culture conditions, but all cells showed high levels of DR antigen after exposure to interferon gamma for 3 days. Similar results were obtained in two independent isolates. We conclude that cryopreserved aortic homografts can contain viable smooth muscle cells capable of expressing major histocompatibility antigens that might render them immunogenic and susceptible to rejection by the recipient's immune system.

Androgens up-regulate the human prostate-specific antigen messenger ribonucleic acid (mRNA), but down-regulate the prostatic acid phosphatase mRNA in the LNCaP cell line

Androgens up-regulate the human prostate-specific antigen messenger ribonucleic acid (mRNA), but down-regulate the prostatic acid phosphatase mRNA in the LNCaP cell line