Human cell line (HGC-27) derived from the metastatic lymph node of gastric cancer

A cell line (HGC-27) was established by culture of the metastatic lymph node from a gastric cancer patient diagnosed histologically as undifferentiated carcinoma. HGC-27 cells were polygonal or short spindle-shaped and adhered to glass surfaces as a monolayer. The cells were probably derived from gastric cancer cells, as their origin from mesenchymal tissues can be excluded morphologically and enzyme-histochemically. Enzyme activities were generally negative or low, except for adenosine triphosphatase, lactic dehydrogenase and leucine aminopeptidase. These scanty findings might reflect the undifferentiated character of the original tumor cells. The cloning efficiency was 5.3% in liquid medium and 1.0% in soft agar. The doubling time was about 17 hr. Chromosomal analysis revealed a mode of 109 and 110 chromosomes.</p

The subendothelial circulatory system in the splenic trabecular vein and the intrasplenic cell recurrence.

In the spleen of patient exposed to atomic bomb as well as in the infective spleen and leukemic spleen sometimes characteristic endothelium of the trabecular vein can be observed and this canalicula in the trabecular vein communicates with reticulum tissue of the pulp. In the subendothelial circulatory canalicula of the splenic trabecular vein there can be observed emigrating picture of various leucocytes of the vein passing this subendothelium (chemotaxisis) and these cells emigrate and accumulate outside the splenic trabecula (intrasplenic cell recurrence). Arterial blood circulates in these subendothelial canaliculae and these canaliculae are not lymph canaliculae as demonstrated by JAGER and ROSSLE. Many leucocytes flow back into the pulp outside the trabecula through this circulatory system. Also in the peritrabecular pulp a new formation of collagen fibers and a considerable number of plasma cells can be observed in various infective spleens, and splenic trabecular area is the regenerating center and reactive center in the spleen, just as lymph follicle in the spleen.</p

Studies on the organellae of liver of cancer bearing animals II. Catalase activity in the liver of animals treated with cyto-plasmic organellae from hepatoma cell (AH 130) and liver cell of the hepatoma bearing animals

The author studied the distribution of polysaccharides and the amino-acid composition of cytoplasmic organellae, the problems that have come to call a great interest in the field of studies on cancer bearing animals. And also biochemical and electron microscopic observations were carried out to study the influences of cytoplasmic organellae in the cancer cells (AH 130), the livers of cancer bearing animals, and normal liver on the catalase activity of the liver. The results obtained are as follows : Cytoplasmic organellae of various cells do not affect so markedly the hexose metabolism of the liver. As for the amino-acid pattern of cytoplasmic organellae of various cells studied by paperchromatography, it is interesting to note that the pattern of the liver of cancer bearing animals, shows lack in histidine, while in can~er tissue and in the liver of cancer bearing animals an increase in phenylalanine can be observed. The decrease in the liver catalase activity is caused by the primary factor of cancer cells, especially their microsomes, and also by the secondary factor of the liver mitochondria in cancer bearing animals. On the other hand, the mitochondria of cancer cells, instead of reducing the catalase activity in the liver, markedly increases the catalase activity. By the morphological changes observed with light microscope and electron microscope, liver cells revealed marked morphological differences, proving that the microsomes of hepatoma cell induce considerably marked changes in the liver, while the mitochondria of hepatoma cell, on the contrary, induce the hypertrophy of liver cells. Sirriilarly in the electron microscopic observations the mitochondria of mouse liver injected with cancer mitochondria are enlarged, but no destruction of cellular structures such as cristae can be recognized. Also microbodies and the growing process of mitochondria can be observed, but no marked changes in endoplasmic reticulum.</p

Studies on the organellae of liver of cancer bearing animals I. Distribution of mucopolysacoharides in the organellae of liver in cancer (Hepatoma AH 130) bearing animals

For the purpose to reveal whether or not the liver and the cell organellae are responsible for the abnormal metabolism of polysaccharides found in cancer bearing individuals, the author analyzed the liver and ascites with tumor cells of AH 130 hepatoma bearing rats biochemically with some histochemical observations. A quantitative increase in polysaccharides accompanied by the production of unusual polysaccharides is found in the supernatant of liver from cancer bearing rats, but not from mitochondrial or microsomal fractions. Tumor cells themselves and ascites fluid do not contain the abnormal polysaccharides found in the liver supernatant.</p

Histochemical studies of adenosine triphosphatase activity of liver cells exposed to ribonuclease

It has been revealed that ribonuclease (RNase) can penetrate into living cells and inhibits amino acid incorporation into proteins resulting in the suppression of protein synthesis and growth of living cells. BHIDE, BRACHET&#185;, KAUFMAN and DAs have proven that the RNase penetrates into onion root-tip cells and induces a number of mitotic abnormalities. KIMOTO and others&#178; also have revealed that RNase injection into mice results in the reduction of cytoplasmic basophilia with the morphologic change of endoplasmic reticulum and the disturbances in DNA synthesis as demonstrated histochemically on pancreatic exocrine cells and liver cells. But there is little information so far on the mechanisms of penetration of RNase into living cells. PILLERI&#179; and SCHUMAKER4 in Brachet's laboratory have demonstrated the uptake of RNase by pinocytosis in amoebae and cancer cells. This may suggest the penetration of RNase through the membrane of the endoplasmic reticulum in the cells whose RNase contents are low5, however it is reasonablly supposed that some phosphatase may be concerned with the permeability of RNase.</p

Cell transformation of established human cell line induced by SV-40 DNA

In the present experiment, it has been noted that clonizing epithelial-like cells of the intestine 407 were more susceptible to SV-40 virus than normal fibroblasts in primary human cell cultures. In the early stage of the infection the cell growth was enhanced by the inoculation of DNA virus but many cells died, showing lysis characterized by CPE, clumping of chromatin and formation of inclusion bodies. On the other hand, the cells surviving infection have given rise to virus-free long term cultures and cellular responses to the virus characterized by cell proliferation which is. classified in four phases. (Phase. I: infection and cell alteration. Phase. II: crisis. Phase. III: fibro-reticulum cell formation. Phase. IV: recovery and proliferation). The most remarkable morphological characteristic was fibroblastic cell alteration from epithelial cells at 5 weeks of virus inoculation. By this study an interesting generalization of human epithelial-like cells can be made about the differentiation of the transformed cells in relation to SV-40 virus and it has been shown that an established human cell line is still susceptible to the reverting action of the SV-40 virus.</p

Oncogenic properties of nucleic acid derived from SV-40 virus

The present report describes the findings on the infectivity of DNA partially purified from SV-40 which was propagated in the monkey kidney cells (BSC-1) in vitro and the importance of nucleic acids as oncogenic factors, particularly the induction of tumor by DNA in newborn hamsters. 593 newborn hamsters in total were used in the present experiments, and cannibalism among them posed as a serious problem. On 30 days postinoculation, very remarkable changes occurred in the liver, lung and subcutaneous areas. Cellular responses of the perivascular cells were predominant. and they were distributed in the interstitial tissues of the liver (liver cirrhosis in primates) and lung. Three hamsters of those subcutaneously inoculated with nucleic acids developed tumors and two tumors appeared in the subcutaneous tissues on 130 days postinoculation, which were identified to be the ones induced by intact SV-40 virus. Other tumors appeared in the liver, lung, intestinal ducts and abdominal surface at 126 days after subcutaneous injection. The cytological observations revealed multiple hemangiosarcoma combined with proliferation of the perivascular cells. On the other hand, cellular responses to nucleic acids were more marked by inoculation of the cell-free filtrate of BSC-1 infected by DNA than of DNA, and essential histologic findings were similar to the respo.nse to infectious DNA. Thirty-nine hamsters (30 per cent) developed tumor within about 200 days postinoculation of the filtrates. Sarcomas were common and they were confined to the subcutaneous tissues in 35 hamsters and to the peritoneum in two others by subcutaneous inoculation of the filtrates. The intestinal gland-cell carcinomas, however, could be induced at 37 and 59 days postinoculation in two hamsters of one litter (7 newborn hamsters) and in the other three newborn hamsters subcutaneous sarcomas were induced by inoculation of the same agent. These results suggest that the observation on the oncogenic capacity of nucleic acids would give us a clue to resolve the course of cancer from the view point of the infectious nucleic acid.</p

Glucose-6-phosphatase activity in regenerating cholangiole cells and hepatoma cells

Histogenesis of hepatic cancer has been analysed by observing glycogen by PAS staining and the histochemically demonstrable G-6-Pase activity on the liver of rats fed with 3'-Me-DAB or 3'-Ni-DAB. By observations on normal hepatic tissue it has been revealed that these two reactions are specific to the cytoplasm of liver parenchymal cells. Observations on the liver from the early stage of dye feeding, up to 100 days, show a marked proliferation of cholangioles in 3'-Me-DAB feeding on polished rice but only a poor reaction of cholangioles in 3'-Me-DAB feeding with synthetic diet. After 15-16 weeks of 3'-Me-DAB feeding cancer develops, a great erpart of which is consisted of cholangiocellular carcinoma and a portion, hepatocellular carcinoma. Histochemical observations on G-6-Pase and glycogen reveal that regenerating cholangiole and adenomatous tissues seem to have poles, on one side, the cells differentiate to liver parenchymal cells and on the other side, they differentiate to bile duct cells. Cancers develop mainly from these regenerating adenomatous tissues and they develop to cholangiocellular cancer or to hepatocellular cancer. The histogenesis of the latter can be traced histochemically. In the cases fed with 3'-Ni-DAB, the activity of cholangiole cells and the development of adenomatous tissue are rather poor with the delayed cancer formation. However, in these cases the majority of cancers are of hepatocellular carcinoma and the developmental mode of hepatocellular cancer can easily be traced by the G-6-Pase activity.</p