THE INHERITABILITY O F SPONTANEOUS TUMORS
O F THE LIVER IN MICE1
STUDIES IN THE INCIDENCE AND INHERITABILITY OF SPONTANEOUS TUMORS IN MICE
SEVENTH
REPORT
MAUD SLYE
From the Cancer Laboratory of the Otho S . A . Spragw Memorial Institute and iAe
University oj Chicago
Received for publication, September 18, 1916
The work of the past six years in this laboratory has definitely
established two points in regard to the behavior of spontaneous
cancer.
I. CANCER IN GENERAL 18 INHERITABLE’
This statement is based upon the following observations:
(1) There are strains of mice in this laboratory which under
no provocation have ever been made to produce spontaneous
cancer.
(2) There are other strains of mice which, under the right
provocation, inevitably do produce cancer.
(3) It is possible by selective breeding to manipulate these
two types of mouse strains with the same certainty with which
it is possible to manipulate pure-breeding pigmented mice and
pure-breeding albinos, and to derive therefrom pure-breeding
cancer strains and pure-breeding non-cancer strains.
I say “cancer in general,” because until one has completely
1 Presented before the American Association for Cancer Research, Washington, D. C., May, 1916.
* For the sake of brevity I shall use the expression “inheritability of cancer,” although cancer is not transmitted as such; rather it is transmitted aa a
tumor-producing potentiality. See introduction to Third Report, Jour. Med.
Research, 1915, xxxii, 159.
503
504
MAUD SLYE
analyzed the reproductive potentialities of every individual concerned in the genealogy of a family, it is impossible to direct
what shall be produced in any given case; and the resulting
strain is likely to show sarcomas or carcinomas, adenomas or
endotheliomas-of the ovaries, or of the mammary gland, or
of the lung, or of the liver, or of any other organ. A family,
therefore, manufactured by the hybridization of cancer-bearing with non-cancer-bearing individuals, will be peppered with
tumors of different organs and of different types. Even
after the cancer-bearing potentialities of two parents have
been analyzed as completely as possible, we still discover in the
offspring some divergence from the ancestral type. From such
experiments carried on through years and in enormous numbers,
yielding year after year perfectly consistent and logical results,
no matter what strains are used, there is but one conclusion
possible, namely, that this tumor-producing potentiality is a
thing indubitably transmitted by the right selective breeding.
11. TUMORS OF SPECIFIC ORGANS AND O F SPECIFIC TYPES ARE
INHERITABLE
That is, by selective breeding it is possible to derive strains
of mice which yield a high percentage of lung tumor or of mammary gland tumor, or of liver tumor, or even of stomach and
uterine tumors which are very rare in mice. And these strains
of mice, again, can be manipulated, carrying into strains with
which they are hybridized the types of tumors borne in their
own ancestral line but from which the family with which they
are crossed is perfectly free. A general survey of this field
appears in the fifth report of these studies published in this
issue.
The present report concerns itself with the inheritability of B
single class of these tumors of specific organs, namely, tumors
of the liver.
The data presented by a single strain or by many strains
carrying a bigh percentage of mammary gland or of lung tumors
when inbred, and introducing a high percentage of tumors of
SPONTANEOUS TUMORS OF THE LIVER I N MICE
505
these same organs into every strain with which they are hybridized, might be contested on the ground of the frequency of these
types of spontaneous tumors in mice, since about 90 per cent
of all reported mouse tumors occur in these organs. Or the demonstration of a relatively high frequency of stomach or of uterine tumors in certain strains might fail to convince, because of
the paucity of these tumors in mice.
But tumors of the liver are of sufficient rarity in the literature to make an interesting study and are sufficiently frequent
in this stock to make a most striking one. In all mouse literature outside of these studies there is but one tumor of the liver
reported, namely, an adenoma reported by the Imperial Cancer
Research Fund of London (Third Report).
This stock to date has furnished sixty-two primary tumors
of the liver. These tumors are chiefly adenomas, with a sprinkling of sarcomas, and of carcinomas, and they are described
fully in the Fourth Report of this series.3 It has furnished seventeen secondary tumors of the liver; these are mostly spindlecell and osteosarcomas, with a few carcinomas. All these primary liver tumors have fallen in strains of identical ancestry,
namely, ancestry derived from Strain 90, and among all the
other cancer-bearing strains, yielding twenty-five hundred primary neoplasms appearing in this stock, not one liver tumor
has occurred. This fact furnishes a most striking piece of
negative evidence for the inheritability of liver tumors.
Chart 1
Strains 280 and 281. These strains were produced by mating
female No. 258, who died of a carcinoma of the lung but who
came of a family carrying a heavy percentage of liver tumors
(see Chart 8, Strain 202), with two brothers from Strain 90,
namely, male No. 363 and male No. 286.
1. Mated with male No. 363, who also had carcinoma of the
lung, she produced a line of which 100 per cent carry tumor,
80 per cent of these tumors being tumor of the lungs.
Jour. Med. Research, 1915, xxxii, 171.
c4
G3
PrnENT
Cucc LUNR
STRAIN -280.
D\ac LUNO
CHART1
STRAIN-28 I.
SPONTANEOUS TUMORS O F THE LIVER I N MICE
507
2. Mated with male No. 286 with an adenoma of the liver, she
produced a line showing three cases of liver tumor in two generations, or 9 per cent of liver tumor. There was in this strain
also an adenocarcinoma of the lungs, an adenoma of the lungs,
and three lung nodules showing pre-adenomatous hyperplasia.
Chart 2
Strains 246 and 246. These strains were produced by mating male No. 286 (parent male also in Chart l),with an adenoma
of the liver, with two sisters of Strain 48. Mated with female
No. 412, who died of acute nephritis, he produced a strain showing over 44 per cent of tumor, with four straight generations of
carcinoma of the mammary gland, three generations of adenoma
of the lung, and one case of “fibro-endothelioma” of the ovary.
Mated with the latter’s sister, female No. 26, with earcoma of
the ovary, liver, kidney and mesentery,4 he produced a line showing tumor in 46 per cent of the individuals, and giving two
generations of primary liver tumors.
Note then, that in both instances where male No. 286 was
mated with a female having a liver tumor, the resulting offspring
showed a high percentage of liver tumor. When the same male
was mated with a female without liver tumor no liver tumor
appeared in the resulting strain although that strain carried a
high percentage of tumor.
Chart 3
Strain $02. This chart is drawn to show both maternal and
paternal ancestry, giving only the fraternities which are concerned in the production of liver tumors. The individuals of
this strain and its“ancestry and of its sister strain, Strain 48
(inbred) show nine cases of liver tumor among thirtyeight individuals, or liver tumor in nearly 24 per cent of the individuals; a
tremendous percentage, considering the rarity of these tumors
4 It is practically impossible in this case to say which of these sarcomas is
primary but i t is probable that the ovarian tumor is primary; hence the liver,
kidney and mesenteric tumors have been classed as secondary.
508
MAUD SLYE
c
3
8
CI
-
m
lu
U
71
5
ep
202
STRAIN
(as.
STRAIN
190
STRAIN
-
I
POLINF
% 772
CARCMGL
360’
UNCERTNN
QX
b
Rn INF
P*P LUNG
ADENOMA
LWER
Q 326
I
6 3227 6 11836
PUL IYF
-X-
Q630Livsa
PVL
CHART3
ANCESTRY
RELIDO-
UNCERTAIN
ACUTE INF
-:SC*RC
s C CYST
~ C n OVAR
n SK
c
I
I
FUERPERAL
Q 165
INFECrlON
UTER~M
STRAIN 90
STRAIN5C
PArrRNAL
PARENT
p
PARENT 6
HEM
65 4 2
SlRAlN - 2 0 2 .
BRWCUO- CAR= M G L
CMCLUNG
FhrrvMoNm METAS
Lu~e.5 & METAS MCD~ASTWUM
6 70
KIDNW
hRc
43
CFRTAlN
UN-
Q5
METASUN
CLLRCMGL
LlVLR
9,348 Q 1199
Q
Q 148
CERTAIN
- ; :
03
Saac M GL
PARENTO STRAIN
90
PARENT6 STRAIN 94
MATERNAL ANCESTRY.
510
MAUD SLYE
in mice. The construction of this strain involved the use of
maternal and paternal ancestry from identical stock, carrying
liver tumor. The potency of female No. 3 to transmit liver tumor
is shown in strain after strain derived from her.
Note here also that female No. 26 when inbred with her brother,
male No. 814, with myocarditis, produced offspring with liver
tumor, just as she did when hybridized with male No. 286 in
Strain 245 (Chart 2). Note then: Female No. 258 (Chart 1)
with lung carcinoma and coming from a strain rich in liver tumor
mated with a male with lung carcinoma produced 80 per cent of
lung tumor; the same female mated with male No. 286 with
adenoma of the liver produced 9 per cent of liver tumor.
Again, male No. 286 with adenoma of the liver mated with
female No. 258, produced liver tumor; mated with female No. 26
of Strain 48 (Chart 3), he produced liver tumor, while mated with
female 412 without liver tumor and coming from a branch of the
family rich in mammary gland tumor, he produced four generations of mammary gland tumor.
Again, female 26 with sarcoma of the liver, outbred as above
with male No. 286, with adenoma of the liver, produced liver
tumor. Inbred with male No. 814, she produced liver tumor.
The assumption of a chance determinant here is absurd where
every individual parent is doubly or trebly checked as to his
potentialities in the matter of liver tumor transmission.
STRAIN 56
0
EDEMA
URINAW ACUTE
LUN~S
RETENTION INFECTION lNFFCTlON
ACUTC
NCCROSII
Aascrrs
LIVER
02
LIVER
AOCNOMA
LIVLR
GI
02
CHART4
SPONTANEOUS !l7JMORS OF THE LIVER I N M I C E
51 1
Chart 4
Strain 69. This is another strain derived from female No. 3,
crossed with a male from Strain 143 who died before autopsies
were being made. Here again she produced offspring with adenoma of the liver. Strain 143 never showed liver tumor.
Chart 6
Strain 616. The parent of this strain, female No. 630, with
metastatic carcinoma of the liver, is shown with her ancestry in
Chart 3, second filial generation. She was crossed with male
No. 721 of an allied line to make Strain 215. The resulting
strain shows three cases of primary liver tumor in three successive generations.
Chart 6
Chart 6 shows a portion of Strain 215 with its parentage for
six generations. The result is seven straight generations of
tumor, three of these involving primary tumor of the liver.
We come now to the consideration of a strain which alone has
produced twelve cases of primary liver tumor, namely, Strain
338. It is a strain manufactured during the last four years from
stock deliberately selected to test the inheritability of tumor
types, with emphasis upon liver tumors.
1. It must be remembered that in work with mice, liver tumor
can not be diagnosed until autopsy. In deliberately breeding to
test the inheritability of liver tumor therefore, one is blind as to
whether or not the individual selected will show it.
2. It must be remembered again that in order to get sufhient
numbers of offspring from the tested individuals for such a rare
tumor as liver tumor, the selected mice must breed young. One
must therefore frequently use mice which have not yet shown
any tumor at all.
3. The worker, then, must be guided by the ancestral history,
and by the available data in the fraternities of the individuals
selected for this test.
In spite of all these handicaps note the results:
G5
STRAIN- 215.
CNEW
Q 9595
CNEPH LOCAL
HIP LIVER
Q I2207
LIVER
Aomomr
WOLMDSUNcrrrrur, UNCEF~TUW
6 7976 68138 6 8139 9 8871
SPONTANEOUS TUMORS OF THE LIVER I N MICE
513
ANCESTRY: STRAIN-215.
SARC
M GL
MALIG
ADENOMA
93
LIVER
UNCERTAIN
6 360
I
-
-n-
I
CARC.M GL
Q METAS
5
LUNQS
PUL.INF
CARCM GL
C NEPH
METAS
LUNGS
I
LUNGS
9 lLIVER
8871
618139
CHART6
Chart 7
Strain 112. The parent female of this liver tumor strain, Strain
338, was female No. 5417. She presented no evidence of liver
tumor but was selected to start Strain 338 after the appearance of
her mammary gland carcin0m.a. She had six litters of young after
the appearance of her tumor and is one of the individuals previously reported (Third Report, p. 193), in whom tumor growth
G3
G PARENT
I
3581
CNEPH
Qi5333 Q 5859
NEPH
CHART7
5156
NECK
Q 5 7 5 8 Q 7690
SS CARC
S a R c LIVER CAac M GL
CNEPH
CARCR i v i s
Q 5417 METASLUNGS
Q 5305
I
WOUNDS CNEPH
Q
PYELO- WOUNDSWOUNDS
Q
639426400664018
6 2512
CNEPH
L ~ O R VNCERTAI UNCERTAIN
Q 1332 Q 1250
LABOR
STRAIN-112
Q3
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or
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6~TIONS
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9
,.., BRANCH
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TWo
uvER
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UV£R
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TlJMOR BEHIND """ENT ~
IGN<PH
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CWO.N 'C
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U 30
6 GCNERAT'IONS __ LIVeR
v.- STRAIN 338.
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ORIN.R[-
.. IUNctATNN
u 842
'--.-----'
IMuLTAas .l..IVEAIADENOMA LIVE!=!:
& 1('0 9 ee3
0 27
OtLATA TlOflI HtA.R t
I
9 24
MeT...............
2e-cM Gt.
CA.rtC F'lOoftS
91 54 17
SUP BtMPrlW..Mfns
L <V<R
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STRAIN
SA OC M GL
MA I..JG AocfWf'tA
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ANCESTRY -
TAPEWOR,..
(lot
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G,uENT
SPONTANEOUS TUMORS OF THE LIVER IN MICE
515
was greatly retarded by constant pregnancy. She lived nearly
a year after the appearance of her mammary gland carcinoma and
showed at autopsy an additional carcinoma of the pelvis and
lungs riddled with metastases. Indeed, at the time of her death
the mouse was about one-half carcinoma.
The place of this female within her inbred strain is shown in
Chart 7, second filial generation. It will be noted that her
grandmother, female No. 3920, had adenoma of the liver, and
that her sister born in the same litter, female No. 5305, had sarcoma of the liver. Inbred with her brother, male No. 3942,
with pyelo-nephritis, she produced mammary gland and neck
carcinoma with metastases in the lungs.
Chart 8
Strain 124. The paternal parent of Strain 338 was male No.
7736. He was selected as the strongest remaining representative of strain 124, which had liver tumor ancestry behind it.
His great-aunt, female No. 1070, had adenoma of the liver.
His immediate family at the time he was selected showed 25
per cent of cancer, but no liver tumor.
He himself lived to be three years and three months old. He
died of arterial sclerosis, and showed at autopsy a papilloma of
the lung.
His place in his inbred strain is shown in Chart 8, Branch 11,
filial generation 3.
Chart 9
Chart 9 is drawn to show the complete ancestry which lay
behind these two parents, namely, female No. 5417and male No.
7736.
Ancestry offemale No. 6417. Five generations back her grandparents were female No. 3, with a malignant adenoma of the
liver, inbred with her brother, male No. 30. with chronic nephritis (these original forbears were members of Strain 90).
The male in her maternal line, and the female in her paternal
line, were the immediate offspring of this pair, the female No.
I
-I-
BRANCH -I
I
STRAIN- 124.
BRANCH -It
SPONTANEOUS TUMORS OF THE LIVER I N MICE
.
517
883 having adenoma of the liver. Hybridization in this generation was with Granby, Mass., red stock, namely, female No. l,
female No. 24 and male No. 27, which never in my hands produced liver tumor, primary or secondary, either in inbreeding or
in hybridization with non-liver tumor strains. Female No. 1,
however, carried carcinoma, and male No. 27 had abscess of the
liver.
The parent female two generations back, female No. 3920, had
adenoma of the liver and the male, No. 3024, had an infected
lung nodule and chronic nephritis.
There were, then, behind the female, three generations of liver
tumor and an ancestry carrying a' considerable percentage of
cancer, with a tendency of the liver to yield to disease. The
origin of both branches of the ancestry was female No. 3 with
an adenoma of the liver which appeared later in both sides of
the family. These facts determined the selection of female No.
5417, who at the time of her selection had a very small carcinoma of the mammary gland and was in excellent breeding
shape.
Ancestry of male No. 7736. The original forebears of this line
also five generations back were female No. 3 with her adenoma
of the liver, and her brother, male No. 30, both long lived.
Again, on both sides of this line also, a double dose of the
immediate offspring of female No. 3 was introduced, namely,
female No. 73 with carcinoma of the lung and male No. 752 with
liver necrosis.
Here as in the maternal line there was a considerable percentage of tumor, a reappearance of adenoma of the liver in the
strain, and a tendency of the liver to yield to disease. There
were then two generations of liver tumor behind male No. 7736,
and three generations of liver tumor behind female No. 5417.
Two adenomas of the liver appeared in the first filial generation
from this cross, namely, female No. 9544 and male No. 8751.
The rest of the chart shows Branch .K,.fr~m,~hjg,q~ss,
bred
out in five lines, the female parent o f . t ~ ~ ~ ~ ~ ~ 8619,
~ ~ ' i ' f i ~ ~ ~
showing two carcinomas 05 ,$?+.qapppv gImd,.a& t+ [email protected]
parent of the branch, m a l e i ~ ~ ; , ~ 7 ~ ~ ~ * ~ ~ * ~ d e n ~ . ~ ~ . t ~ l i J e r .
..............
. . . . ..........
. -.: .
..........
:..::*:.
:,*
*.a.
518
MAUD SLYE
In the second filial generation there were three tumors of the
liver, namely, one sarcoma and two adenomas.
In the third filial generation also there were three liver tumors, all adenomas.
Note that these liver tumors all fell within two lines from this
mating, namely, lines A and D. Note also line C, of which
100 per cent shows carcinoma, all of the mammary gland and of
the lung; where the parent female had carcinoma of the mammary gland and the parent male had carcinoma of the lung.
Chart 10
Branch IX of Strain 338. This branch is derived, of course,
from identical ancestry with Branch V, as shown in Chart 9.
Here also there are two generations of primary liver tumor.
Chart I 1
Branches XI1 and XI11 of this same strain show also an outcropping of primary liver tumor in the fourth filial generation, a
lymphosarcoma in female No. 12212, and an adenoma in female
No. 11245.
Chart 12
Parts of Strains 465 and 460. This chart is introduced to
show how in hybridization also Strain 338 and its sister strain,
Strain 465, produced liver tumor.
Two sisters, female No 5305 and female No. 5417, were mated
with two half-brothers, male No. 5215 and male No. 7736. Note
the outcropping of liver sarcomas, along with the stomach tumors,
in the third and fourth filial generations in each case.
Let me review the striking points demonstrated by these
charts :
1. A great many strains have been carried for years in this
laboratom.&lfi@g .yaying percentages of cancer, some of them
as highi.%&,$Qp&x@&ti .$$..never showing liver tumor, primary
.oy.$econdaqy., 9These.strqips h v d a e e n carried side by side with
.the;%$& $,$qbr, . & t r a , i i ~ b j$$&$andled
$
with identical tech-
..............
... ...:. . . . ...:. .:. :.
..............
*.:
a.:
B
W
G.3
G.z
G.1
b 9518
~ADENOMAL I V ~ .
ADENOMA LUNG
G.PARENT
B R ~ N Cx
H
? 12386
[SITE
OF
WOUNDl
S Q C L L L C A R CFACE
z CARC.M.GL
CHART10
? 11828
SARCOMA
THYMUS.
METAS.DIAPHRAGM
9 12172
PNEUMONIA
KIDNEY
CYSTS.
8 68.34
SCLEROSIS.
TAPEWORM
ARTERIAL
PAPILLOMALUNG
-1
~NK,~NF.
338
0 12138
STRAIN
63
G.2
G. i
G.PARENT-
CHART12
~NTUTlNAL~NFtCTl
OCCURREN
Srorucrr TUM
%OWING
n c . L u ~ADENC++ALWCR
~.
PARTS O f STRAINS 4 0 5 AN0 480 WITH ANCCSTRX
522
MAUD SLYE
nique, and hybridized with identical non-liver tumor strains, thus
eliminating every possible cause for this divergence except
heredity.
2. One cancer bearing strain, Strain 90, has been used either in
inbreeding or in hybridization (a) in the origin of every strain in
this laboratory which has ever produced a primary tumor of the
liver; (b) in the origin of every strain which has ever furnished a
secondary tumor of the liver, with the exception of two osteosarcomas which metastasized in practically every organ in the
body.
3. One individual female, No. 3, with a malignant adenoma of
the liver and a sarcoma of the mammary gland, stands out preeminently in this production of liver tumor. Either she or her
immediate offspring has been used in one or both origins of
every strain which has ever produced a liver tumor, primary or
secondary, in this laboratory, with the exception of the two
metastatic osteosarcomas noted above.
The emergence of liver tumor in these hybrid strains derived
from female No. 3, or her immediate offspring, frequently takes
place years after the death of their liver tumor progenitors,
thereby eliminating every possibility of a contact transmission
of this disease even in the nature of a germ plasm infection.
4. Certain individuals, both when inbred anti when outbred
and when tested with many mates, consistently show liver
tumor in every resulting strain.
Consider (1) that outside of this stock there is just one liver
tumor recorded among all the thousands of mouse tumors, (2),
that these results are deliberately produced by the manipulatioii
of selective breeding alone, (3) that they have consistently occurred for years in every test made, (4) that there are over
twenty-five hundred primary neoplasms, and over fourteen thousand individuals involved in this perfect and persistent consistency. What explanation remains but that of heredity?