C-of Ets-2, C-myc And N-ras Antisense Rna Reversed On Hepatoma Cells And Anti-human Hepatoma Monoclonal Antibody Development,

Human hepatocellular carcinoma (HCC) is one of the most highly prevalent malignant tumors in China. Unfortunately, because the onset of symptoms coincides with advanced stage of the disease, the prognosis is generally poor. The mortality is very high. The advent of monoclonal antibody (McAb) technology has allowed new possibilities of HCC diagnosis and even therapy. Over the last ten years, a large number of McAbs with the potential clinical application have been raised. It remains a major goal to identify and raise new hepatoma selective McAbs with minimal or no normal tissue crossreactivity, in order that McAb could be used as a potentially important tool in the battle against hepatoma. In the present investigation a McAb (B11) was raised against human hepatoma cell line 7721 which seemed to bind more or less specifically to human liver carcinoma cells. A immuno-conjugate of epirubicin with the McAb was prepared and its cytotoxicity to hepatoma cells in vitro was studied.Balb/c mice were immunized with human hepatocellular carcinoma cells. Spleen was removed and splenocytes were fused with Sp2/0 myeloma cells using 50% PEG. The culture supernatants were assayed by ELISA and thirty positive clones were obtained. One hybridoma was obtained with high specificity for hepatoma. After being frozen and thawed for several times, the hybridoma remained the capability to secrete antibody. Karyotyping was performed and it revealed that the hybridoma had ninety to ninety-eight chromosomes. Immunoglobulin isotype analysis suggested that the B11 was of IgG2b immunoglobulin isotype. The titer of the ascites fluid of B11 was assayed by ELISA to be between 105 ~ 106. The specificity of the antibodies was assessed by binding studies to a panel of malignant and nonmalignant human cell lines, with ELISA and methods. The results demonstrated that the B11 also showed crossreactivity with two human colon carcinoma cell lines and one human breast carcinoma cell line, but not with eleven other human tumor cell lines and one normal human cell line, including three pancreatic carcinoma ell lines, three leukemia cell lines, one gastric carcinoma cell line, one laryngocarcinoma cell line, one esophageal carcinoma cell line, two cervical carcinoma cell lines and. the human embryonic lung fibroblast cell line. On the other hand, the reactivity of B11 was...