| Background and AimsThe basis for genes research, tumor cytogenetic research contributes to the studies of tumor associated genes and inherited genes. It is an important method for carcinogenesis research to cloning potential genes according to recurrent chromosomes aberrations of tumors. Chaoshan is located in esophageal carcinoma (EC) high-risk area of south China, where most people came from other EC high-risk areas of China. EC in this region is charactered by the different incidence between sex, high -risk families, high incidence for emigrants. It is suggested that genetic factors might play a part in the etiology of EC in this region. Because of the difficulty to make direct chromosome preparations from primary rumor tissue, the establishment of EC cell lins is important for cytogenetic research about EC. To get some cytogenetic information of EC in this region for molecular genetic research and to provide material for therapeutic research, a cell line derived from primary EC in Chaoshan region was established. Its biological features including chromosome aberrations were studied too.Material and Methods1 Source of Tumor Specimen. Twenty-seven EC specimens for primary cultures were obtained from the affiliated tumor hospital of Shantou University Medical College. Just one EC cell line from a 46 years old female in Chaoshan region with well-differentiated carcinoma in the middle esophagus was established successfully.2 Primary Culture and Establishment of CSEC-215 cell line. Primary cultures of EC cells were initiated by tissue explantation or treatment of collagenase II.3 The following biological characters were studied, ﹎orphologic characteristics and ultrastructure of the cell line observed by inverted microscope, light microscope, electron microscopy, ゞrowth kinetic features as doubling time, mitosis index, the efficiency of forming clones on dishes and soft agar, cell cycle, attachment rate and cultures with low saturation serum.﹖umorigenicity to Scid mice with double immunity system deficiency, 〆xpression of keratine , p53 and HPV protein detected by immunohistochemistry assay.4 Cytogenetic Features. The chromosome number was counted for 80 and 100 metaphases of passage 9 and 45, respectively. Karyotyping analyses were made for twelve metaphases of each passage.Results1 Primary Culture and Establishment of CSEC-215 cell line. Theprimary cultures reached confluence and were subcultured for the first time after they had been cultured in vitro for 24 days. They had grown continuously in vitro for more than nine months and subcultured 66 times.2 Morphologic Characters and Ultrastructure. The cell line grew as monolayer and piled up in high saturation densities. The cells were pleomorphic with the majority being polygonal in shape and with bundles of tonofilaments in cytoplasm.3 Growth Kinetic Features and Immunohistochemical Stain. The doubling time of the cell line ranged from 25.7 to 39.6 hours according to the number of cell inoculated, with the maximum mitosis index 44%. The efficiency of forming colonies more than 50 cells on dishes was 14%. It failed to grow in soft agar and media without serum, but grows well in media with 2% or 5% serum. The attachment rate within 24 hours was over 90%. The cells survived of passage 8 were 70% after they had been conserved in liquefied nitrogen fore over half a year. The cells cycle showed that 77.3% of cells at GO+G1 phase, 18.9% at S phase, 3.8% at G2 and M phase respectively. The cell line had tumorigenicity to Scid mice and the histological features of tumor developed were similar to the origin EC with ultrastructural demonstration of tonofilaments and desmosomes. The immunohistochemistry stain indicates that the xenografts and the cellline expressed keratin. The origin EC and the cell line also expressed the p53 and HPV protein.4 Cytogenetic Features. CSEC-215 cell line was found to be aneuploidy. The modal number of passage 9 was not obvious, while...
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