Hidehito Ichihashi

Characteristics of three human gastric cancer cell lines, NU-GC-2, NU-GC-3 and NU-GC-4.

Three human gastric cancer cell lines, NU-GC-2, NU-GC-3 and NU-GC-4 were establishedin vitro from the cancer tissues obtained from 3 patients during surgery. The pathological findings of the gastric tumors of these cases revealed poorly differentiated adenocarcinoma (and partial signet-ring cell carcinoma in the case of NU-GC-4). NU-GC-2 and NU-GC-4 were originally obtained from metastatic paragastric lymph nodes and NU-GC-3 was obtained from a metastatic tumor in the brachial muscle. The cells of NU-GC-2 and NU-GC-3 are polygonal in shape and grow as a monolayer sheet. NU-GC-4 cells, however, are mainly spherical in shape with a few free floating cells. Electron microscopy revealed epithelial characteristics in all 3 cell lines. The average doubling time of NU-GC-2 was 36.1 hours, that of NU-GC-3 was 38.2 hours and that of NU-GC-4 was 29.9 hours. The modal chromosome number of NU-GC-2 was 62, that of NU-GC-3 was 58 and those of NU-GC-4 grown inin vitro andin vivo were 52–54 and 53, respectively.In vitro andin vivo lines of NU-GC-4 were established from the same tumor. These two cell lines are quite similar in morphology, but slightly different in karyotype. Thein vitro sensitivity to anticancer agents was highest in NU-GC-4 and lowest in NU-GC-2. Of the anticancer agents, mitomycin C and adriamycin were most effective on the cells of all 3 cell lines.

Liver regeneration and tumor growth in the rat after partial hepatectomy

Tumor growth of Yoshida sarcoma implanted in the remnant liver was studied in rats subjected to a hepatectomy. After 70 percent hepatectomy, the liver progressively regenerated and the total liver weight was reverted to be 10 days after the operation. Concomitantly with liver regeneration, tumor growth in the remnant liver was stimulated significantly, compared with that in the sham-operated liver. Incorporation of tritiated thymidine into tumor cells in the remnant liver was strikingly high and progressive, while that in the sham-operated liver was low and retained. Mitomycin C given to the hepatectomized rats was more effective against the tumor in the remnant liver than in the sham-operated liver. We conclude from this study that cancer cell proliferation in the remnant liver can be accelerated by the process of liver regeneration.

Application of Radioactive Precursors for the Evaluation of Sensitivity of Cancer Cells to Anticancer Drugs

An in vitro drug sensitivity test was developed to estimate the lethal effects of drugs on cancer cells. Cancer cells were incubated in the presence of radioactive precursors and anticancer drugs. The drug effects were estimated from the changes in rates of incorporation of precursors into DNA, RNA, and protein. A microtiter plate and a multiple automatic cell harvester made feasible handling of a large number of samples. Incorporation of radioactive precursors was well correlated with drug-induced cell lethality. The results of this test were also correlated with in vivo regression of tumors of mice. This test appeared to be more reliable than other similar tests of cell lethality in vitro. For utilization in clinical studies, a test plate was simplified and 25 human cancers were tested. The in vitro results demonstrated a positive correlation with clinical results in 80% of the observations.

Esophageal cyst, a case report and a review of the literature.

The esophageal cyst is a rare disease only 20 cases of which have been reported in the literature in Japan. We recently treated such a patient and at thoracotomy, we found the cyst to be located the submucosal layer of the esophagus. The cyst could be easily extirpated and the patient made an uneventful recovery. Gross findings revealed thick, yellow and mucus-like contents inside the cyst. Microscopically, the cyst was lined with ciliated columnar epithelium and there was no evidence of cartilage. The 21st such occurrence in Japan is reported herein and a discussion is made of related literature.

The effect of intravenous hyperalimentation on cell mediated immunity.

The effect of intravenous hyperalimentation (IVH) on cell mediated immunity was examined in 22 patients. Each patient received PHA and PPD skin tests before and after the performance of IVH. In this study both PHA and PPD skin reactivity showed significant increase after IVH, and serum albumin levels had positive correlation with the PPD skin reaction changes.Absence of the established delayed hypersensitivity in the surgical patient, especially those with malignant diseases, is probably secondary to generalized malnutrition, and established cell mediated immunity can be restored by proper nutritional repletion.

Esophago-pleuro-cutaneous fistula a case report

In a sixty-three year old Japanese man with a history of long standing pulmonary tuberculosis, an unusual esophago-cutaneous fistula developed. The possibility of esophago-pleuro-cutaneous fistula was considered, because there was an old tuberculosis causing lung abscess or hilar lymph node adenopathy and which facilitated development of an extensive fistulous tract. The patient was effectively treated by palliative surgical procedure. This may be the first report of a benign esophagopleurocutaneous fistula.

A simple and rapid in vitro test for determining sensitivity of cancer cells to anticancer drugs.

For practical utilization of chemosensitivity tests of malignant cells to anticancer drugs in clinical studies, we developed a new method which is simple, rapid, and applicable to fresh human tumors. The principle of the method is to measure the content of ATP of living cells by chemoluminescence, after drug exposure. A straight linear relationship was observed between the number of viable cells and the light intensity. This method seems to be a good indicator for the selection of anticancer drugs in cancer chemotherapy.

Glomus tumor of the stomach.

A case of glomus tumor of the stomach, found 7 cm oral to the pylorus of a 23-year-old man was reported.

A stomach oncofetal antigen recognized by monoclonal antibody GC302

A monoclonal antibody, GC302, was established by fusing murine myeloma NS/1 cells with the splenocytes of a BALB/c mouse immunized with a human gastric cancer cell line, NU-GC-3. The serological specificity of GC302 was analyzed by an anti-mouse Ig mixed-hemadsorption (MHA) test on a panel of human cell lines, and an immunoperoxidase method using the frozen sections of tumors and normal tissues of adult and fetus. GC302 reacted with cancers of the stomach and colorectum but did not react with hepatocellular carcinomas, melanomas, or astrocytomas in the MHA tests. By the immunoperoxidase method, GC302 was found not to react with normal adult gastric mucosa, but to react with the mucosa in the fetal stomach, intestinal metaplasia, and almost all of the cancer of the stomach. GC302 also reacted with the normal mucosa of the intestine, colon, and rectum as well as with cancers of these origins. In normal liver sections, the antibody reacted with the bile ducts, but not with the hepatic cells. These results indicate that the antigen detected by GC302 is characterized as an oncofetal antigen in the stomach, and also as a differentiation antigen whose localization discriminates between the gastrointestinal tracts of the forgut origin and those of the midgut and hindgut origin. The molecular weight of the GC302 antigen was estimated to beca. 40,000 by the Western blot analysis. Periodic acid treatment on the antigen suggested that the antigenic determinant is a carbohydrate.

Correlation of an in vitro chemosensitivity test using [3H] incorporation with the response in case of human tumor chemotherapy.

We developed a method for performingin vitro drug testing using [3H] incorporation into a primary human tumor. The test was applied to specimens from 60 patients with advanced malignancies, mainly gastrointestinal cancers. Forty-six specimens had sufficient growth for the drug testing. Forty-seven per cent of the gastric cancer specimens and 67 per cent of those from patients with colorectal cancer demonstrated anin vitro chemosensitivity. Among 9 chemotherapeutic drugs tested, ara-C, 5-fluorouracil, and mitomycin C showed the high percentages of positive sensitivity for gastric cancer, whereas actinomycin D, carboquone, and nimustine hydrochloride were sensitive in case of colorectal cancer. Clinical responses were compared with the results of the test. In 46 drug assays,in vitro/in vivo drug responses correlated in 78 per cent, with a true-positive rate of 47 per cent and a true-negative rate of 94 per cent. This test appears to be a reliable indicator for the clinical response to the same chemotherapeutic drugs in cases of the malignant diseases seen clinically.