Acta Medica Okayamaousar.lib.okayama-u.ac.jp/files/public/3/31408/20160528023459883093/... · acta med. oknynma 15, 9-26 (1960) studies on the organellae of liver of cancer bearing

Acta Medica OkayamaVolume 15, Issue 1 1961 Article 2

FEBRUARY 1961

Studies on the organellae of liver of cancerbearing animals II. Catalase activity in theliver of animals treated with cyto-plasmic

organellae from hepatoma cell (AH 130) andliver cell of the hepatoma bearing animals

Tetsuo Kimoto∗

∗Okayama University,

Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.

Studies on the organellae of liver of cancerbearing animals II. Catalase activity in theliver of animals treated with cyto-plasmic

organellae from hepatoma cell (AH 130) andliver cell of the hepatoma bearing animals∗

Tetsuo Kimoto

Abstract

The author studied the distribution of polysaccharides and the amino-acid composition of cy-toplasmic organellae, the problems that have come to call a great interest in the field of studieson cancer bearing animals. And also biochemical and electron microscopic observations werecarried out to study the influences of cytoplasmic organellae in the cancer cells (AH 130), thelivers of cancer bearing animals, and normal liver on the catalase activity of the liver. The resultsobtained are as follows : Cytoplasmic organellae of various cells do not affect so markedly thehexose metabolism of the liver. As for the amino-acid pattern of cytoplasmic organellae of vari-ous cells studied by paperchromatography, it is interesting to note that the pattern of the liver ofcancer bearing animals, shows lack in histidine, while in can∼er tissue and in the liver of cancerbearing animals an increase in phenylalanine can be observed. The decrease in the liver cata-lase activity is caused by the primary factor of cancer cells, especially their microsomes, and alsoby 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, markedlyincreases the catalase activity. By the morphological changes observed with light microscopeand electron microscope, liver cells revealed marked morphological differences, proving that themicrosomes of hepatoma cell induce considerably marked changes in the liver, while the mito-chondria of hepatoma cell, on the contrary, induce the hypertrophy of liver cells. Sirriilarly in theelectron microscopic observations the mitochondria of mouse liver injected with cancer mitochon-dria are enlarged, but no destruction of cellular structures such as cristae can be recognized. Alsomicrobodies and the growing process of mitochondria can be observed, but no marked changes inendoplasmic reticulum.

∗Copyright c©OKAYAMA UNIVERSITY MEDICAL SCHOOL

Acta Med. Oknynma 15, 9-26 (1960)

STUDIES ON THE ORGANELLAE OF LIVER OF CANCERBEARING ANIMALS

11. CATALASE ACTIVITY IN THE LIVER OF ANIMALSTREATED WITH CYTOPLASMIC ORGANELLAEFROM

HEPATOMA CELL (AH 130) AND LIVER CELLOF THE HEPATOMA BEARING ANIMALS

Tetsuo KIMOTO

Department of Pathology, Okayama University Medical School, Okayama,(Director: Prof. S. Seno)

Received for publication, June 10, 1960

Already J. P. GREENSTEIN1.2·3,4,ll and K. NAKAHARA6•7 pointed out the

existence of a specific substance in the cancer tissue, toxohormone, that reducesthe catalase activity in the liver. Nakahara and co-workers7 reported that thisdiminution factor of the catalase activity exists in microsomes of cancer cells. In1956 E. WEILER8 verified the difference between the cancer cells of the liver andnormal liver cells by tracing the antibody with fluorescent dye in his antigenantibody reaction tests conducted with the microsomal fraction of normal rat liveras the antigen. Thus, the role played by ctyoplasmic organellae in vivo is gainingmore and more biological significance. As for the antigen contained in cytoplasmic organellae the polysaccharide is important, and the author has alreadypointed out the localization of this polysaccharide distributed in each cytoplasmicorganella. H. MASAMUNE and S. HAKOMORI clarified the existence of k-mucopolypeptide in the urine of cancer bearing patients and suggested that cancercells, by secreting some unknown substance (primary substance) in the entireorgans, change the function or the template of the cells to produce mucoproteinsspecific to each visceral secretion into a cancerous state.

With the purpose to study the influence of the cytoplasmic organellae ofthe normal liver and those of cancer and tumor bearing animal livers on normalliver, the author carried out cytochemical and electron microscopic studies onthe catalase activity other than .polysaccharides, and also the amino acid composition of various cytoplasmic organellae that are so closely associated withthe catalase activity as well as the changes brought about in the liver after theadministration of the cytoplasmic organellae of different composition, in aminoacids and the previously reported polysaccharides.

9

1

Kimoto: Studies on the organellae of liver of cancer bearing animals II.

Produced by The Berkeley Electronic Press, 1961

10 T. KIMOTO

MATERIALS AND METHODS

Isolation of Cytoplasmic Organellae :

Same as in the previous report, the fractions such as the cell supernatants,microsomal and mitochondrial fractions are isolated by Schneider's method fromAH 130 cells (ascitic and solid hepatoma), the liver of the AH 130 bearing ratand from normal rat liver. These fractions are dialyzed. The filtrate or theaseptic distilled water suspension prepared with the frozen-dried powder areused as materials. One half cc of the filtrate or the suspension is injected intraperitoneally into a group of pedigree strain mice repeatedly for more than 7days up to 20 days. As the control, normal mouse liver, distilled water, andphysiological saline solution are injected similarly into the mice. The experimental groups are consisted of ten animals each. The livers of these animalsare used for the determination of the catalase activity and for the quantitativeanalysis of hexose. As for the microscopic observation the livers are fixed in a10 per cent formalin solution and alcohol. They are stained with PAS, hematoxylin-eosin (H-E), and carmine. For the electron microscopic observation, thetissue is fixed with a two per cent osmic acid solution.

The distribution of polysaccharides and the amino acid composition of thecytoplasmic organellae of the rat livers to be used for the injection are analyzedby means of paperchromatography. The HCI eluate obtained after adsorptionon Dowex 50 as illustrated in previous report is put in a warm dessicator toeliminate HCI, and then it is thoroughly washed with distilled water and dried.This dried residue is again dissolved in 0.3 cc distilled water and 0.03 cc of thissolution is spotted. For the primary development (8 hours) n-butanol, glacialacetic acid and water (4 : 1 : 2/vol), and for the secondary development (14hours) 1/10 N borax and phenol (1: 10/vol), and the coloration is reacted withninhydrin.

The catalase activity of the mouse liver in the experimental groups is estimated with Battelli-Stern's apparatus. Namely, after sacrificing the mice bydecapitation, 100 mg liver is put into the Waring blendor containing 10 ccphosphate buffer solution, pH 7.6 and is homogenated one minute. Next, themixture of 0.3 cc homogenate and 0.7 cc phosphate buffer is placed in the flaskof Battelli-Stern's apparatus, and to this mixture 3 per cent H202 is added andthe flask is shaken for five minutes. Oxygen gas evolving is measured gasmetrically, at 10-15°C, under the pressure of 760 mm (Hg). Taking the catalseactivity of the normal liver as 10, the catalase activity under 7.5 is consideredto be the reduced value.

For the determination of the changes in the hexose content and for thequalitative analysis of polysaccharide composition of the mouse liver by paper-

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Acta Medica Okayama, Vol. 15 [1961], Iss. 1, Art. 2

http://escholarship.lib.okayama-u.ac.jp/amo/vol15/iss1/2

Liver of Cancer Bearing Animal 11chromatography, the liver homogenate in the frozen-dried powder is used. Asexplained in the previous report the quantitative analysis is conducted in thesame way with the water eluate after the adsorption on Dowex 50 and also thequalitative analysis of polysaccharide is performed by the paperchromatography.

RESULTS

A considerable vanatIon in the amino acid composition can be found ineach of organellae such as the cell supernatants of cancer cells (AB 130), thelivers of cancer bearing animals, and normal liver, microsomal, mitochondrialfractions, that are to be used for the injection. From the amino acid patterns,most of amino acids can be found in the cell supernatant fraction, followed bythe microsomal fraction (Tables 1, 2). Table 3 illustrates the composition of aminoacids in each organella. As can be observed from. this table, no histidine canbe detected in the livers of the cancer and cancer bearing animals. This provesto be the sole characteristic that is common to both the hepatic cancer cells andthe cancer bearing rat liver as far as the amino acid composition is concerned.Furthermore, in the liver of cancer bearing animals, phenylalanine is increasedmarkedly as compared with other fractions.

The catalase activity of the mouse liver after the injection of these organel.lae coincides with the histological findings of the liver to be described later.Namely, the marked decrease in the catalase activity is the same as observed inthe liver of cancer bearing rats (Table 4), that is, both the liver of the miceinjected with cancerous microsomes and the liver of the mice treated withthe mitochondria of cancer bearing rat liver show as low as 6.0. In thecontrol group the liver of the mice injected with physiological saline solutionshows the value of 8.0, revealing only a relatively slight decrease as comparedwith the normal untreated liver. This value is approximately the same as thecatalase activity of the livers of the mice injected with the cell supernatant, microsomes or the mitochondria of the normal rat liver. On the contrary, themouse liver after the injection of the mitochondria of the hepatoma cell showsthe activity rather higher than the normal value, and histologically and electronmicroscopically, as will be described later, a marked swelling of the cells canbe recognized. As can be seen from these findings, it is obvious that cytoplasmicorganellae, especially in cancer cells and cancer bearing liver, bring about astriking change of the catalase actvity in the liver.

However, in the determination of the hexose content in the livers of themice injected with various cytoplasmic organellae there can be recognized nogreat change in the value as shown in Table 5. In the qualitative analysis ofpolysaccharides analyzed by paperchromtography, glucose and ribose can bedetected in the liver of the mice injected with hepatoma cell supernatant;

3



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VAL HEr~ IJD-tElI MET!o [/AL VAt In'o0 0 TYr<. 0 0 .ALA. irK ALA. ALA. Tr~.

o 0o fHR. .-'-. 0THR 0 THR-;J "0 :. : 0 <flY

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Cell SPN of hepatoma cell Cell SPN of cancer bearing rat liver Cell SPN of normal rat liver

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. ~ !.Table 1. Variation in the amino acid composition of each cell supernatant

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microsomes of cancer bearing rat liver

N-BUTANOL, GLACIAL ACETIC ACID & WATER(4: 1: 2/VOL)

Table 2. Variation in the amino acid composition of each microsome.

microsomes of hepatoma cell

-OARf.

OLY~ .PHEO· ~.

0lQ

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Q 0 L8-~

o°TffR.

/fD-lEV.H~T~ /f~-{E{)

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14 T. KIMOTO

Table 3. Common composition of amino acids in hepatoma cell and cancer bearingrat liver.

I NORMAL LIVER I AH 130 BEARING RATS I AH 130 CELLSLIVER, CELL. IMICRO-I CELL.

IMICRO-

ICELL. I MICRO-SPN SOMES SPN SOMES SPN SOMES

t PHENYLALANINE II

+[; ....... ~

.. I.. ::i.::.::::J +I

I HISTIDINE l + I I I ] II

10 r-

CONTROL! EXPERIMENTAL MICE GROUPSI

Table 4. Catalase activity of mice liver injected with cytoplasmic organellae

6



Liver of Cancer Bearing Animal

Table 5. Hexose contents in the livers of the mice injected with variouscytoplasmic organellae.

15

CONTROL I NORMAL LIVER I 11.5% II

CELL. SPN 7.9

AH 130 CELLS MICROSOMES 10.0MICE LiVERS

INJECTED WITH MITOCHONDRIA 10·7CYTOPLASMICORGANELLAE

CELL. SPN 8.4AH 130 BEARING

MICROSOMES 10.0RAT LIVER

MITOCHONDRIA 17.0

glucose, mannose, and ribose in the case injected with cell supernatant of cancerbearing rats; glucose, mannose, and ribose in the case injected with the mitochondria of hepatoma cells; and mannose and ribose in the case injected withthe mitochondria of hepatoma bearing rat liver as shown in Table 6, but nomarked differences can be observed in the composition and qualitative analysisof polysaccharides.

MICROSCOPIC FINDINGS

In the liver of the mice injected with the supernatant of hepatoma cell theperipheral area of acini show a diffuse degeneration in a moderate degree, and inthe liver with the decreased catalase activity the cytoplasma has become transparent, showing vacuolar degeneration, in addition. The glycogen and PAS stainsappear diffusely in the liver lobules, generally on the periphecrial area of acini.

In the mouse liver after the injection of cancer bearing rat liver cell supernatant, vacuolar degeneration can be seen scatteringly, but on the whole thedegeneration is slight. By glycogen and PAS stainings no marked changes canbe recognized. In the liver of the mice injected with normal liver cell supernatant the findings are rather similar to these findings just mentioned.

In the liver of the mice injected with cancerous microsomes, the degeneration of cytoplasma is marked, shown caryorrhexis, necrosis as well as adecrease in the substances positive to glycogen and PAS stains (Plate 2). Onthe contrary, in the liver of the mice Injected with microsomes of the cancerbearing animals no degeneration of the liver cytoplasma can be recognized, andthe liver cells around blood vessels are rather swollen, showing striking basophilia in the cytoplasma but no marked changes in the substances positive toglycogen and PAS stains.

In the case of the group injected with the mitochondria of cancer cells, theswelling and enlargement of the liver cells are most marked among the experi·

7



16 T. KIMOTO

Table 6. Paperchromatographic finding of mice livers injected with variouscytopla.,rnic organllae.

GALACT= GALACTOSE. MAN =MANNOSE. L. ARAB=L. ARABINOSE.XYL=XYLOSE GLUC = GLUCOSE. RIB = RIBOSE.

ffif-<~

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0 0 0 0 0 0 0...l

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:iMO 0 0 ~ 0.....

Cl MAN i MMJ..... MANP:::>t lARAB 0 010 0 0 0p..

0 (fLUC (TWC IGLUC GiVC ~LU( ~LVC

~AlACTI

I

mental groups, paralleling with the rise in the catalase activity (11.0-15.0),but the degeneration can hardly be observed. Moreover, the liver cytoplasmacontains basophilia and most of liver cells are hypertrophic (Plate 3, in the samemagnification as Plate 4, notice the hypertrophy of liver cells). And also nomarked changes can be recognized in the substances positive to glycogen andPAS stains. On the other hand, in the group injected with the mitochondria ofcancer bearing rat liver, the mouse liver shows a diffuse degeneration of thecytoplasma and a marked vacuolar degeneration. This finding parallels with thedecrease in the catalase activity (Plate 4). The substances positive to glycogen-

8



Liver of Cancer Bearing Animal 17

and PAS stains are diffusely increased.In the groups injected with microsomes and with mitochondria of normal

rat liver both show slightly a diffuse vacuolar degeneration (Plate 1).

ELECTRON MICROSCOPIC FINDINGS

The liver of AH 130 cancer bearing rats shows a fairly marked degeneration and transparency of cytoplasmic organellae such as endoplasmic reticulum(ER) and mitochondria, but in a portion of the nuclear membrane pore, sphericalor tubular ER are arranged densely and are seen migrating toward cytoplasma(Plate. 5).

The liver of the group injected with cancerous micorsomes shows the disintegration and agglomeration of microsomes and in addition mitochondria aremarkedly swollen and these cristae are extremely distorted (Plates. 7, 8). Especially in the liver of the group injected with microsomes of cancer bearing livermarked agglomerations of Palade's granules and intracellular substance can beobserved diffusely as illustrated in Plate 9. A portion of ER shows vacuolation,and .besides these, mitochondria are generally normal or electron dense, revealing fat droplets in places.

The livers of the group injected with the mitochondria of the cancer cellsshow the swelling of mitochodria. However, as can be seen in Plate 10 theswelling and the increase of mitochondria in number are marked, and no destruction of cristae can be recognized. Some of the mitochondria are extremely irregularin size, and the number of microbodies of Roueller and Bernhard are increased.In the group injected with mitochondria of the cancer bearing liver some ofPalade's granules are agglomerated anP the degeneration, disintegration, anddegeneration of mito~hondria can be observed while some of ER's are markedlyvacuolated (Plate 11).

DISCUSSION

In his investigations of the catalase activity in rat hepatoma and ]ensen'ssarcoma mice ]. GREENSTEIN1

.2.3.4,ll (1942) reported that the decrease in thecatalase activity is due to a marked diminution in the production capacity ofliver catalase and suggested the cause of such a decrease might be attributed tothe presence of some inhibitory substance. K. NAKAHARA6

•7·(1948) on the other

hand, considered "toxohormone" to be the responsible factor for the dereasein the catalase. More recently D. ADAMS (1950) et al. reported that thedecrease in the catalase activity is brought about not only by tumor but alsoby a kind of stress, castration, and hypophysectomy, etc. S. T AKAHARA9 madesome case report of "Aeatalasemia". NAKAHARA and ONO pointed out thatbiologically 'toxohormone' activity exists only in microsomal fraction of cancercells.

9



18 T. KIMOTO

The author carried out biochemical and pathological investigations on theliver functions in cancer bearing animals and studied the biological changes inthe liver brought about by cytoplasmic organellae of cancer cells. The polysaccharide composition of cytoplasmic organellae in various cells is already reportedin the previous paper, but since 'toxohormone' is supposed to be a polypeptideas claimed by K. NAKAHARA et al., following upon the polysaccharide composition, by means of paperchormatography the author studied the amino-acidcomposition that is so closely associated with toxohormone. Regarding aminoacids in cancer, it is said that the amino acid pattern of the tumor bearing liverdoes not differ from that in normal liver according to B. SCHWEIGERT10 (1940),M. SANBERLICH and C. BAUMANNll (1951), but H. LEVY and M. DUNN12

(1953) reported that amino acids in tumor bearing liver are mostly free aminoacids because of autolysis, while M. DUNN13 (1949) found a slightly greateramount of arginine and threonine, M. MICKELSON and N. BARVICK14 (1956)more asparagic acid than in the normal liver. Besides these, there are interestingworks on amino acids in connection with cancer by such investigators asE. ROBERTS and P. BoRGES16 (1955), ]. PRICE16 et al (1948) and V. WARAVDEKAR and O. POWERS17 (1957).

In the author's paperchromatographic studies on the cytoplasmic organellaeof various cells such as cancer cells (AH 130) and cells of cancer bearing animalliver and normal liver, in the hepatoma cells it has been found that histidineis markedly decreased or is absent as illustrated in Tables 1, 2, 3. Coinciding with this finding, in the liver of AH 130 cancer bearing animals amarked decrease or absence of histidine can be recognized. Needless to say thatit requires still more precise biochemical studies on these points, but the relationbetween the cancer growth and amino acids, the histidine metabolism in particular, seems to be an important problem for future studies. On the other hand,it is interesting to note that in the liver of cancer bearing animals phenylalanineis increased, and the attention need be called on the importance of the cancerbearing animals and the phenylalanine metabolism. At the same time pathologically the relationship between the adrenal function and the pancreas functionseems to be taken into account.

Coupled with the previous report on the polysaccharide composition, cytochemical and electron microscopic studies were conducted concerning the influences of cytoplasmic organellae with such an amino acid composition asmentioned above on the livers. In this instance an attention must be called firstof all to the liver catalase activity as shown in Table. 4. From this a special attention is again called to the fact that the factor inducing the decrease in the livercatalase exists not only in cancerous microsomes but also in the mitochondria ofthe liver of AH 130 cancer bearing rats. Those livers showing a decrease in

10



Liver of Cancer Bearing Animal 19catalase all revealed a marked degeneration of mitochondria, electron microscopically. Likewise ER and Palade's granulues were disintegrated and agglomerated in the liver of the group injected with cancerous microsomes(Plates 7,8).In the liver injected with the mitochondria of cancer bearing animals as shownin Plates 11, mitochondrion is markedly degenerated. On the contrary, in themitochondria of cancer cells themselves, there is a substance that accelerates theliver catalase activity above the normal level. In the injection of cancerousmitochondria the liver cells are hypertrophied markedly and basophils appearabundantly. Electron microscopically likewise, the mitochondria are markedlyswollen, but cellular structures such as cristae are not injured and mitochondriaare rather increased in number, and microbodies can be observed numerously,presenting a picture of the mitochondrial growth or production process (Plate 10).

From these findings as observed by NAKAHARA, of the various phenomenaobservable in the cancer bearing animals the changes occurring in the cytoplasmic organellae and in the liver catalase activity seem to be considerablyspecific and important. However, the author calls attention to the factor inducing the decrease in the catalase activity is also contained in the liver mitochondriaof cancer bearing animals, in addition to the cancerous microsomes, the factorautomatically inducing the decrease in the catalase activity of cancer cells themselves as pointed out by Nakahara. In other words, the decrease in catalaseactivity of liver occurring in the cancer bearing animals is due to the metabolitesof the mitochondria of the cancer bearing liver (secondary principles), causingthe modification in the synthesis of the toxohormone-like substance, apart fromthe decrease caused by cancer cells themselves (primary principle), to be exactby microsomes of cancer cell.

CONCLUSION

The author studied the distribution of polysaccharides and the amino-acidcomposition of cytoplasmic organellae, the problems that have come to call agreat interest in the field of studies on cancer bearing animals. And also biochemical and electron microscopic observations were carried out to study theinfluences of cytoplasmic organellae in the cancer cells (AB 130), the livers ofcancer 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 hexosemetabolism of the liver.

As for the amino-acid pattern of cytoplasmic organellae of various cellsstudied by paperchromatography, it is interesting to note that the pattern of theliver of cancer bearing animals, shows lack in histidine, while in can~er tissueand in the liver of cancer bearing animals an increase in phenylalanine can be

11



20 T. KIMOTO

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 factorof the liver mitochondria in cancer bearing animals.

On the other hand, the mitochondria of cancer cells, instead of reducingthe catalase activity in the liver, markedly increases the catalase activity.

By the morphological changes observed with light microscope and electronmicroscope, liver cells revealed marked morphological differences, proving thatthe microsomes of hepatoma cell induce considerably marked changes in theliver, while the mitochondria of hepatoma cell, on the contrary,' induce thehypertrophy of liver cells.

Sirriilarly in the electron microscopic observations the mitochondria of mouseliver injected with cancer mitochondria are enlarged, but no destruction ofcellular structures such as cristae can be recognized. Also microbodies and thegrowing process of mitochondria can be observed, but no marked changes inendoplasmic reticulum.

ACKNOWLEDGEMENT

I wish to thank Prof. Satimaru Seno, Department of Pathology, Okayama University,Prof. Hajime MASAMuNE and Asist. Prof. Senichiro HAKOMORI, Department of Biochemistry,Tohoku University, for valuable direction in the course of this work. My thanks are also dueto Toshiko TOKUDA, Nobuo HAYASHI for technical asistance of electron microscope and Prof.Haruo SATO, Fukushima Medical University for gifts of AH 130 hepatoma strain.

REFERENCES

1. J. P. GREENSTEIN, W. V. JENRETTE and J. WHITE.: The liver catalase activity of tumorbearing rats and the effect of extirpation of the tumors. J. Nat. Cane. Inst. 2, 283, 1941.

2. J. P. GREENSTEIN.: Titration of the liver catalase activity of normal and of tumor bearingrats and mice. J. Nat. Cane. Inst. 2, 525, 1942.

3. J. P. GREENSTEIN and H. B. ANDERVONT.: The liver catalase activity of tumor bearingmice and the effect spontaneous regression of certain tumors. J. Nat. Cane. lnst. 2, 345,1942.

4. J. P. GREENSTEIN. : Note on the liver catalase activity of pregnant mice and of mice bearinggrowing embrynic implants. J. Nat. Cane. Inst. 4, 283, 1943.

5. J. P. GREENSTEIN.: Further studies of the liver catalase activity of tumor bearing animals.J. Cane. Inst. 3, 397, 1943.

6. K. NAKAHARA.: Toxohormone; A characteristic toxic substance produced by cancer tissue.GalUl. 40, 45, 1949.

7. K. NAKAHARA.: Toxohormone biosynthesis. Gann. 45, 1954.8. E. WEILER.: Die Anclerung cler serologischen Spezifitat von Leherzellen cler Ratte Wahrend

cler cancerogenene durch P-Dimethylaminoazobenzol. Z. Naturforschg. 116, 31, 1956.9. S. TAKAHARA.: Acatalasemia. Jibi-Inko-ka. 2\, 53, 1949 (in Japanese)

10. B. S. SCHWEIGERT, B.T. GUTHNECK, J. M. PRICE, J, A. MILLER, E.C. MILLER.: Amino

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Liver of ,Cancer Bearing Animal 21

acid composition of morphological fractions of ,rat livers and induced liver tumors. Proc.Soc. Exp. BioI. Med. 72, 495, 1946.

11. H. E. SANBERLICH and Co A. BAuMANN.: The amino acid content of certain normal andneoplastic tissues. Cane. Res. 11, 67, 1951.

12. H. M. LEVY, E. A. MURPHY and M. S. DUNN.: Effect of ethionine on tumor growth andliver amino acids in rats.' Canc. Res. 13, 507, 1955.

13. M. S. DUNN.: The amino acid composition of a fibrosarcoma and its normal homologoUstissue in rat. Cane. Res. 9, 306, 1949.

14. M.MICKELSON and L. BARVICK. : Some biochemical differences in nonnal and tumor tissues(sarcoma ISO) of mice; Amino acid composition. J. NatCane. Inst. 17, 65, 1956.

15. E. ROBERTS and P. BORGES.: Patterns of free amino acids in growing and regressingtumors. ,Cane. Res. 15. '697, 1955.

16. J. M. PRICE, E. C. MILLER and J. A. MILLER.: The intracellullar distribution of protein,nucleic acids, riboflavin and protein·bound aminoazo dye in the livers of rats fed P-Dimethylaminoazbenzene. J. BioI. Chem. 173, 345, 1948.

17. V. WARAVDEKAR and O. POWERS.: Reduction of synthesis of diphosphopyridine nucleotidein tissues from mice bearing transplanted tumor. J. Nat. Cane. Inst. 18, 145, 1957.

Explanations for Photos

Plate 1. The mouse liver of the group injected with the microsomes of normal liver: H-Estain, high magnification, showing a slight vacuolization of liver cytoplasma.

Plate 2. The mouse liver of the group injected with cancerous microsomes: H-E stain, highmagnification. The liver cytoplasma is highly degenerated (V), showing nuclear disintegration and necrosis, at the same time a decrease in the substance positive to PASstain.

Plate 3. The mouse liver injected with mitochondria of hepatoma cells (AH 130):H-E stain, high magnificatin. Hardly any degeneration can be observed in liver cytoplasma. On the contrary, liver cells are hypertrophied and the nuclei are enlargedat the same time basophils is rich in cytoplasma. (in angle. C: Cytoplasma. N: nucleus: cf. Plate 5).

Plate 4. The mouse liver of the group injected with mitochondria of the cancer bearing ratliver: H-E stain, high magnification.The degeneration of liver cytoplasma and vacuolization are marked. (V)

Plate 5. The liver of AH 130 cancer bearing rat.The nuclear membrane shows ER and its particles on its pore. And cytoplasma isstrikely transparent. (T) N: nucleus. NM; nuclear membrane. M: mitochondria.ER: endoplasmic reticulum.

Plate 6. The mouse liver of the group injected with microsomes of normal rat liver:A degeneration of mitochondria (M) can be recognized, but electron density of RNAgranulues is increased (D). ER: endoplasmic reticulum

Plate 7,8. The liver of the mouse injected with cancerous microsomes of AH 130 hepatomacells.

The enlargement and degeneration of mitochondria can be seen. (M) Most of cristaeare destroyed, showing a marked vacuolization (V) and revealing the so-called fatdroplets. (F) N: nucleus.

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22 T. KIMOTO

Plate 9. The liver of the mouse injected with microsomes of the cancer bearing rdt liver:Palade's granulues and intracellular substances are diffusely agglomerated, (G) andthe mitochondria are relatively normal. M: mitochondria. E. R: endoplasmic re·ticulum, F: fat.

Plate 10. The mouse liver of the group injected with the mitochondria of AH 130 cells.Mitochondria are swollen and cristae are distinct. (M) In this plate a fairly goodnumber of fine mitochondria can be seen and some of them resemble regeneratingmicrobodies. (MB) ER and Palade's granulues are distinct. (ER) N: nucleus.

Plate 11. The mouse liver of the group injected with the mitochondria of cancer bearing ratliver. A marked vacuolization of ER can be observed in cytoplasma. (V) Some ofmitochondria are shapeless, degenerated and atrophied. (M) E. R: endoplasmic reticulum.

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Liver of Cancer Bearing Animal 23

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24 T. KIMOTO

16



Li\"er (){ L'anl'er Bearing :\nim;d 25

,t:i·~,.

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Acta Medica Okayamaousar.lib.okayama-u.ac.jp/files/public/3/31408/20160528023459883093/... · acta med. oknynma 15, 9-26 (1960) studies on the organellae of liver of cancer bearing