STUDIES ON ENZYME ACTION
XXXVII. A FURTHER STUDY OF THE COMPARATIVE LIPASE
ACTIONS OF HUMAN TUMORS
K. GEORGE FALK AND HELEN MILLER NOYES
From the Harriman Research Laboratory, The Rooaevelt Hospital, New York
INTRODUCTION
The results obtained in the study of certain enzyme actions
of a number of tumors of human origin were presented in a
previous paper.' In this a number of additional results will be
considered which make clearer the differences between the
characteristic enzyme actions of some of the tumor types and
which will enable a more definite formulation of the further
problems involved to be made.
EXPERIMENTAL METHODS
The experimental methods were essentially the same as those
described in the former paper. The tumor material was obtained from the Pathological Laboratory of the Roosevelt
Hospital. The writers wish to thank those who furnished the
material for this study and for the ready cooperation which
they extended at all times. These thanks are due especially to
Dr. C. W. Lester and the Interne Staff of the Hospital.
Since the enzyme technique was the same as before, it may
be described briefly. The tumor material was ground and
extracted with water. The hydrolytic actions of the extracts
were tested on ten different esters, and the amounts of acid
produced in each case under the same standard conditions
measured. All the usual precautions necessary in enzyme experiments were taken.
EXPERIMENTAL RESULTS
A large mass of material bearing upon the problem has been
collected. A number of individual experiments were presented
1
H. M. Noyes, K. Sugiura, and K. G. Fa&, J. Cancer Research, 1925, ix, 105.
146
STUDIES ON ENZYME ACTION
147
in the former paper. In place of presenting more such data
here, only the averages of the various types of behavior found
lc
FIQ.1. FIBROMYOMA
OF UTERUS
EXTRACTS.TYPEI. Average.
OF UTERUS
EXTRACTS.TYPE11. Average.
FIFJROMYOMA
UTERINE
MUSCLE
EXTRACTB.
Average.
will be given and such remarks bearing upon the individual
relations as may appear necessary.
148
K. GEORGE FALK AND HELEN MILLER NOYES
Of the two methods of presenting the results, that based upon
the relative actions on the different esters of a given material
presented in the form of a curve, and that using the absolute
enzyme actions found in the experiments, the former appears to
give the more valuable information, although for a satisfactory
understanding of the relations, both methods muat be used.
Perhaps the most interesting group of the tumors which were
studied included the fibromyoma of the uterus. Two distinct
types were observed whose enzyme pictures were entirely different and not reflected in the histological findings. A considerable number of such fibromyomas have since been studied.
I n place of giving the individual results, averages of the two
types of enzyme actions will be shown.
Results will be included for 71 cases of fibroids which included also six specimens of uterine muscle. 163 separate fibroid
masses were tested. Of these, 55 were found to be similar and
were grouped as Type I as a matter of convenience; 91 were
similar and grouped as Type 11. 17 specimens showed pictures
intermediate between Types I and I1 and will be considered
briefly later. In Figure I are shown therefore the averages of
the relative actions first of the 55 Type I fibroids, second of the
91 Type I1 fibroids and third of the 6 of uterine muscle.
The differencea in the three curves are very definite. Without
entering into details, it is obvious that, with Types I and I1
fibroids, a comparison of the actions of various pairs of esters
would indicate striking differencea. The question of the absolute actions of these fibroid and muscle extracts must be considered. These actions for a given type of material even for the
same concentration vary considerably for any one ester. At
the same time the extremes of the actions found for the two
types and the uterine muscle wiIl be given for the concentration
of 44.4 mg. tissue per cc. of final solution tested in order to
indicate the differences which may be expected as a rule.
Although there is considerable variation in the actions on
any one ester, it is clear from Table I that the actions of the
Type I1 fibroids are uniformly higher than those of the Type I
fibroids. The differences summarized here and in the curves of
149
8TUDIES ON ENZYME ACTION
.I
PhOAc. . . . .
Gl(0Ac)s. . . .' .' .' .' .
MeOCOPr., . . . . .
PhCHiOAc . . . . . . .
%ltOAc.
.
.....
_.
MeOAc. . . . .
EtOCOPr.. . . . . . .
MeOBz. . . , . . . . . .
EtOBz. . . . . . . . . .
i-BuOAc .
.....
I
.
.
.
.
FigPoids
Type I1
Fibroid6
Uterine
Muscle
0.50-0.93
0.21-0.62
0.10-0.41
0.00-0.18
O.OPO.18
0.09-0.28
0.02-0.33
0.00-0.08
0.00-0.03
0.00-0.26
0.78-1.38
0.44-0.91
1.16-2.71
0.07-0.44
0.41-1.29
0.50-1.64
1.02-2.75
0.20-0.70
0.20-0.65
0.03-0.53
1.20-1.47
0.72-0.88
1.45-1.99
0.23-0.35
0.55-0.90
0.66-1.11
1.33-2.00
0.32-0.70
0.31-0.62
0.26-0.36
I n the results which are averaged and summarized here, 71
sets of specimens including 163 fibroid masses were studied.
Of these sets of specimens, 16 showed Type I actions only, 28
Type I1 actions only, and 5 Intermediate Type actions only.
22 sets of specimens showed more than one type of action for the
different masses present. Of these, 13 sets showed Types I and
I1 actions, 2 sets Types I and Intermediate, 3 sets Types I1 and
Intermediate, and 4 sets Types I, 11, and Intermediate. These
results show a fairly general distribution of the different types
of fibroid actions. In the previous paper, with the limited
number of results then available, no connection was observed
between the enzyme type of action of a fibroid mass and its
histological structure or physical characteristics. The same
fact was found with the larger mass of material for which the
results are presented in this paper.
The comparative actions of the Type I1 fibroids and the
uterine muscles may be considered briefly. The limits of the
absolute actions of these two sets of materials shown in Table I
do not show any definite differences. However, the curves of
150
K. GEORGE FALK AND HELEN MILLER NOYES
the relative actions are such as to show real differences in type.
I n the former paper, too few results were at hand for the
uterine muscle to bring out such a difference clearly. At the
FIG.2. BLADDER
TUMORS.EXTRACTS.
Average.
FIBROMYOMA
OF UTERUS
EXTRACTS.
Type I. Average.
same time it must be borne in mind that these uterine muscle
results were obtained with material from operations; that in
every case fibroid masses were present imbedded in the uterus
wall; and that therefore this uterine muscle material cannot be
considered unequivocally as normal tissue.
STUDIES ON ENZYME ACTION
151
The results of the bladder tumors, a number of which were
presented in the earlier paper will be summarized. Eight specimens were studied. The averages of the relative actions are
shown in Figure 2 together with the averages of the Type I
uterine fibroids.
!4C
190. 3. LIPOMAS.EXTRACTS.Average.
FIBROMYOMA
OF UTERUS
EXTRACTS.Type 11. Average.
152
K. GEORGE FALK AND HELEN MILLER NOYES
There is a general similarity in these curves, although the
Type I fibroids show higher general actions with phenyl acetate
as standard. The histological findings indicated in general
various types of malignant growths. Some of these were given
in the previous paper. The concentrations of the extracts
tested varied considerably so that it is impossible to consider
the absolute actions.
The averages of the relative actions of three lipomas are
given in Figure 3 together with the averages of the Type I1
fibroids.
The results for the lipomas agreed very closely so that the
average may be assumed to give greater significance than would
be indicated by the few results obtained. Although there is a
general similarity to the Type I1 fibroids there are at the same
time differences which appear to be real and not of accidental
nature.
DISCUSSION
The results presented here confirm and extend the conclusions
of the earlier paper. The two types of fibromyoma of the uterus
are clearly characteristic in their enzyme behavior. Compared
with other types of human tumors, there appears to be a general
resemblance between Type I and malignant bladder tumors
and Type I1 and benign lipomas. At the same time, there are
real differences to be recognized between the two in each group.
Uterine muscle was found to be definitely different in picture
from Type 11, although they approach each other much more
than the Type I and the uterine muscle. A number of enzyme
pictures (17 out of 191) were obtained with uterine fibroids
which indicated a type intermediate between Types I and 11.
Not enough constant results were obtained to permit of general
conclusions, but several possibilities may be indicated. Some
of the results may have been due to experimental errors. With
fibroids showing small absolute actions, several errors of 0.2
to 0.3 cc. in the titrations might account for the difference from
Types I or 11. While such errors may account for some of the
results, they cannot account for all. It is possible that both
types of actions are present in the same fibroid mass and that the
STUDIES ON ENZYME ACTION
153
enzyme result found shows the sum of these actions. Finally,
it is also possible that the results represent the true nature of
the fibroid masses in question, and that in addition to the two
extreme types, I and 11, there are real intermediate types ranging
from the one extreme to the other.
The results in this paper have been limited to the tumor
types which either on the basis of location or of histological
finding, may be grouped together. A number of isolated results,
isolated in the sense that they refer to different tissues, were
presented in the former paper. While the enzyme pictures
found with them show general similarities, they cannot be
grouped as yet in a satisfactory manner.
SUMMARY
The study of the hydrolyzing actions of extracts of human
tumors on ten esters has been continued. The results obtained
with a considerable number of uterine fibroids, bladder tumors,
and lipomas, are summarized and presented. Two distinct
types of uterine fibroids are again shown. The uterine muscle
actions differ from both but are much more like one of these
types than the other. This type of uterine fibroid resembles
the lipomas in the enzyme actions, although distinct differences
may be observed. Similarly, the other type of uterine fibroids
resembles the bladder tumors, but again with certain differences.
The relations are best brought out by means of the graphs
showing the relative enzyme actions on the different substrates
by one group of tumors, and comparing the different graphs or
“pictures.” The absolute enzyme actions of some of these
tumor materials are also indicated.
ERRATA
In an articIe by George L. Rohdenburg and Otto F. Krehbiel
entitled “On the Relation of Certain Endocrine8 to the Salt
Content of Rat Blood,” published in the Journal of Cancer
Research in 1925 (volume x, page 423), there are a few arithmetical errors in the percentage columns of the tables. The
figures in the first four columns are correct. One of these
errors has led t o an incorrect statement at the end of the article
concerning the percentage of calcium in the blood of animals with
an incised wound. It is requested therefore by the authors
that the last sentence of the article be deleted from subscribers’
copies.
In the four articles on “Studies in Biological Resistance to
Malignant Tumors,” by E. B. Krumbhaar and others of the
Philadelphia General Hospital, that appeared in the September
number of this journal, pp. 298-328, a Statement should have
been included that they were aided in part by a grant from the
Elizabeth Thompson Science Fund.
154