Matilde Olive

Long-term human breast carcinoma cell lines of metastatic origin: Preliminary characterization

SummaryNineteen human breast carcinoma cell lines have been established as continous cultures during the past 6 years in our laboratory. This preliminary report is designed to list the lines by their designated code numbers (MDA-MB) and present a brief summary of their morphological, cytogenetic and biochemical characteristics. Sixteen of our lines were obtained from pleural effusions, two from brain metastases, and one from pericardial fluid. All lines have been shown to be distinct entities and are uncontaminated by HeLa cells or each other. A lq marker chromosome is present in all but one of the lines examined.

Characterization of the DiFi Rectal carcinoma cell line derived from a familial adenomatous polyposis patient

SummaryThe DiFi human colorectal cancer cell line was recently established from a familial adenomatous polyposis patient with extracolonic features characteristic of the Gardner syndrome. These cells have now been propagated for 150 passages in standard culture media and vessels without feeder layers or collagen coatings. They retain features of colonic epithelial cells such as surface microvilli, secretory vesicles, and desmosomes. Cytosol of DiFi cells contains a high level (502 U/mg protein) of the mucin CA 19-9. In addition, DiFi cells produce carcinoembryonic antigen, and induce tumors in athymic mice. Cytoskeleton analysis of DiFi cells by fluorescence microscopy showed a pronounced disorganization of actin cable structure. The isozyme genetic signature of DiFi cells is unique (0.01 probability of finding the same genetic signature in a different cell line), differs from that of HeLa cells, and has expressional features seen in other colorectal cell lines. The DiFi cell karyotype is tetraploid, contains many marker chromosomes, and shows numerous episomal particles. Two copies of chromosome 18 were absent, and only a single normal chromosome 17 was found. This parallels detection of allelic losses from DiFi cell DNA at loci on chromosomes 17p and 18 using molecular (cDNA) probes. DiFi cells clearly express transcripts for the c-myc proto-oncogene, the c-myb proto-oncogene, and thep53 tumor suppressor gene. Transforming growth factor beta inhibits DiFi cell growth in soft agar and suppresses c-myc expression in these cells. The value of this cell line in the study of genetic alterations in colorectal cancer is discussed.

Pancreatic adenocarcinoma cell line, MDAPanc-28, with features of both acinar and ductal cells

SummaryConclusionWe established a new human pancreatic adenocarcinoma cell line, MDAPanc-28. Studies on this new line indicate that it expresses both acinar and ductal gene products suggesting that the patterns of gene expression in the pancreatic adenocarcinoma from which this cell line arose have features similar to those of the protodifferentiated cells hypothesized by Rutter and his colleagues for the developing pancreas (1,2).BackgroundThe cell line arose from a tumor that, like most pancreatic adenocarcinomas, was ductal on the basis of its histological appearance.MethodsOnce the cell line was established in culture, they were subjected to cytogenetic analysis and tested for their ability to grow in nude mice. RNA from the cells was analyzed by Northern blot analysis and PCR of reverse transcribed cDNA for the expression of both acinar and duct cell gene products. DNA was analyzed for the presence of mutated K-ras at codon 12.ResultsThe cell line expressed trypsin and ribonuclease RNA, which are considered to be acinar cell markers, and carbonic anhydrase II (CAII), which is considered to be a duct-cell markers. The histological appearance of xenografts in nude mice was similar to that of the tumor from which the cell line was established. The chromosome number varied between 46 and 60.

Human cell line from an adenocarcinoma of the ampulla of vater

Dear Editor: The ampulla of Vater in the duodenal wall is formed by fusion of the main pancreatic duct of Wirsung and the common bile duct (1). Surgeons and pathologists have attempted to separate malignancies of this region based upon the presumed epithelial cell of origin (ampulla, pancreas, bile duct, or duodenum). Virtually all retrospective studies have suggested an improved survival for nonpancreatic ampullary adenocarcinoma when compared with pancreatic duct cell adenoearcinoma (3,5,6,8). The numbers of cases are too few to permit careful stratification for degree of differentiation; presence of vascular, perineural, and lymphatic invasion; lymph node positivity; tumor size; and adequacy of surgical margins. However, the available clinical evidence does support a fundamental difference in metastatic potential between ampullary and pancreatic adenocarcinoma, although their patterns of metastasis are similar. The mechanisms responsible for these differences in biologic behavior are unknown, and investigative efforts at the molecular level are just beginning. The present study represents the first report of a cell line, MDAAmp-7, isolated from an adenocareinoma of the ampulla of Vater. A 38-year-old man developed epigastrie discomfort in October 1987. These symptoms progressed, and jaundice was noted in February 1988. A computed-tomogram scan of the abdomen demonstrated extra hepatic biliary obstruction. Exploratory laparotomy in March 1988 revealed a mass in the ampullary region, and intraoperative biopsy demonstrated adenocarcinoma of the ampulla. The tumor was not reseeted; however, the biliary obstruction was bypassed with a choledochoduodenostomy. Following an uncomplicated postoperative recovery, the patient was referred to The University of Texas M.D. Anderson Cancer Center in May, 1988. Staging evaluation revealed no evidence of metastatic disease and an uncomplicated pancreaticoduodenectomy was performed. A papillary tumor, 1.5 >( 1.5 )< 0.5 cm, was present in the ampulla of Vater with partial occlusion of the lumen of the ampulla and the distal pancreatic duct. Histologically the carcinoma displayed a spectrum ranging from well-differentiated glandular structures to small clusters of cells and highly anaplastic single cells with occasional signet-ring configuration transmurally invasive through the museularis propria of the duodenum into peripanereatic adipose tissue (Fig. 1). Perineural and lymphatic invasion was seen and metastatic carcinoma was present in three of four peripancreatic lymph nodes as well as on the serosal surface of the common bile duct. In February, 1989, the patient developed a nodule in his abdominal scar, and fine-needle aspiration demonstrated metastatic adenocarcinoma. At the time of full-thickness abdominal wall excision of the tumor the presence of multiple small peritoneal implants was demonstrated. The MDAAmp-7 cell line was cultured from a metastatic abdominal implant that arose from the primary adenocarcinoma of the ampulla of Vater.

The DiFi Rectal Carcinoma Cell Line from a Familial Adenomatous Polyposis Patient: An In Vitro Model System to Study Genetic Changes in Colorectal Cancer

A new human colorectal cancer cell line (DiFi) has been established from a patient with the Gardner Syndrome. Characterization studies have shown that DiFi cells secrete carcinoembryonic antigen (CEA) and CA 19-9, are tumorigenic in nude mice, have a tetraploid karyotype, and possess an isozyme pattern characteristic of colorectal cancer cell lines. Southern blot analysis shows that DiFi cells have allelic loss on chromosomes 17p and 18, plus amplification of the EGF receptor gene. Northern blot analysis reveals that transforming growth factor (TGF)-alpha, c-myc, and p53 transcripts are clearly expressed in these cells. The significance of these molecular changes in relation to mechanisms leading to a selective growth advantage in DiFi cells will be discussed.