With this study we have demonstrated that in vitro transduction of normal human CD4(+) T lymphocytes with NPM-ALK results in their malignant transformation. The transformed cells become immortalized and display morphology and immunophenotype characteristic of patient-derived anaplastic large-cell lymphomas. These unique features, which are strictly dependent on NPM-ALK activity and expression, include perpetual cell growth, proliferation, and survival; activation of the key signal transduction pathways STAT3 and mTORC1; and expression of CD30 (the hallmark of anaplastic large-cell lymphoma) and of immunosuppressive cytokine IL-10 and cell-surface protein PD-L1/CD274. Implantation of NPM-ALK-transformed CD4(+) T lymphocytes into immunodeficient mice resulted in formation of tumors indistinguishable from patients' anaplastic large-cell lymphomas. Our findings demonstrate that the key aspects of human carcinogenesis closely recapitulating the features of the native tumors can be faithfully reproduced in vitro when an appropriate oncogene is used to transform its natural target cells; this in turn points to the fundamental role in malignant cell transformation of potent oncogenes expressed in the relevant target cells. Such transformed cells should permit study of the early stages of carcinogenesis, and in particular the initial oncogene-host cell interactions. This experimental design could also be useful for studies of the effects of early therapeutic intervention and likely also the mechanisms of malignant progression.

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Alveolar rhabdomyosarcoma is a pediatric soft-tissue tumor that is often difficult to distinguish from other small round-cell tumors. The PAX3-FKHR and PAX7-FKHR gene fusions that result from chromosomal translocations in this tumor provide potential molecular diagnostic markers. To apply these molecular markers to commonly available archival material, we used reverse transcriptase-polymerase chain reaction and oligonucleotide hybridization methodology to develop an assay capable of identifying PAX3-FKHR and PAX7-FKHR fusion transcripts in formalin-fixed, paraffin-embedded tissue. Use of a control assay for wild-type FKHR mRNA indicated that RNA was successfully isolated, reverse-transcribed, and amplified in 15 of 16 archival cases. Comparison of assay results for the PAX3-FKHR and PAX7-FKHR fusions with standard molecular assays of paired frozen material revealed that all eight cases of known fusion-positive rhabdomyosarcoma were correctly identified and distinguished as PAX3-FKHR or PAX7-FKHR. The seven cases of known fusion-negative rhabdomyosarcoma showed no evidence of either product. These results indicate that we have developed a molecular assay that accurately identifies the fusion transcripts characteristic of alveolar rhabdomyosarcoma in archival samples. This assay will be useful for diagnosis and for retrospective clinicopathologic correlative studies.

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Thirty-four cases of esophageal cancer were treated by radiation combined with chemotherapy and hyperthermia (triple therapy) from September 1985 to January 1986. Hyperthermia was performed by an intracavitary microwave applicator of 915 MHz. The temperature at the tumor margin was 43 degrees C to 44 degrees C, whereas at the middle luminal surface of tumor it reached at 44 degrees C to 48.5 degrees C. Chemotherapy was with bleomycin (PYM) 20 mg/session, intramuscularly (IM) and cisplatin (DDP) 1.0 to 1.5 mg/kg/session intravenously (IV). Hyperthermia and chemotherapy were given simultaneously within 30 minutes after 500 cGy of radiation. Three modalities were given on the same day once a week for six sessions. The total response rate (complete + partial response [CR + PR]) was 94% (32/34) and 1-year and 2-year survival rates were 74% (25/34) and 44% (15/34), respectively. Among them were 23 patients primarily treated by triple therapy with the 1-year and 2-year survival rates of 83% (19/23) and 48% (11/23), respectively. No serious side effects were seen. The average microwave power consumption during the hyperthermic treatment and its prognostic significance are discussed.