January 1979
89
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
ALTERATION OF HUMAN SERUM RIBONUCLEASE ACTIVITY IN MALIGNANCY
Authors: Dalia Maor. Michael R. Mardiney, Jr.,*+ Section of Immunology and Cell Biology Baltimore Cancer Research Center Baltimore, Maryland
Referee:
Paul 9. Chrctien Tumor Immunology Section Suriay Branch National Cancer Institute Bethcsda. Maryland
INTRODUCTION The association of elevation of alkaline Serum ribonuclease activity and cancer noted by several investigators more than two decades ago'.' was clouded by both the absence of elevated activity in all cancer'." and the occurrence of high levels of enzyme activity in a variety of unrelated. nonneoplastic diseases.'-" These results together with a lack of knowledge concerning the source, nature, and biologic significance of ribonuclease activity lead only to a few systematic attempts to determine the usefulness of this finding in clinical oncology. Since then, improved methods for the analysis of serum ribonuclease activity utilizing a battery of synthetic polynucleotides as substrates instead of RNA or a single substrate have yielded greater reproducibility and made possible the detection of subtle changes in enzyme activity not previously noted. This review will summarize those observations which demonstrate the association of alteration of ribonuclease activity in malignancy and will suggest that multiple patterns of change in ribonuclease activity exist within the cancer state. These consist mainly of the elevation of ribonuclease ac-
EVIDENCE DEMONSTRATING THE ASSOCIATION BETWEEN INCREASED SERUM RIBONUCLEASE ACTIVITY AND CANCER Most studies concerning the alteration of serum ribonuclease activity in cancer patients have compared either enzyme activity among normal cancer patients and patients with nonmalignant diseases-the level of serum ribonuclease activity pre- and post-cancer therapy which included either surgery, irradiation, or chemotherapy done or in combination or the level of serum ribonuclease activity among patients in remission, relapse, or those considered cured (nonrecurrence for 5 years or more posttreatment). In 1955 Aleksandrowicz presented the first evidence that there were variations in ribonuclease activity in the urine of leukemia patients.' He noted in further studies that the
Currrrrt add-: 2 Rav-Ashi Street. Rmet-An'v, Israel. Cumnt addnu: How& County Docton' Bldg., 9380Baltimore Natl. Pike. Ellicott City, Md.21043. ~
**
tivity in the serum as well as nonneoplastic tissue and the depression of ribonuclease activity in the tumor cell itself. A possible explanation of these observations will evolve during this review.
~~
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90
CRC Critical Reviews in Clinical Laboratory Sciences
serum ribonuclease activity of patients with chronic granulocytic leukemia was several times higher than that noted in normal individual^.*^1' Also in 1955, Metais tested ribonuclease activity in the serum of 60 young normal males and females, 20 patients with a variety of diseases (tuberculosis, chronic pancreatitis, liver disease, heart disease, and kidney disease), and 16 hospitalized patients with neoplasia.* He found that three patients with pancreatic carcinoma had elevated serum ribonuclease activity while all others studied were in the normal range. In 1958,two groups of investigators reported on alterations in serum ribonuclease activity in cancer. Migliarese tested 75 normals, 48 untreated cancer patients, 34 patients undergoing therapy for carcinoma, and 26 patients with nonmalignant d i ~ e a s e Serum .~ ribonuclease activity was significantly elevated in the untreated cancer patients and either low or within the normal range in patients treated with irradiation or chemotherapy. Only one nonneoplastic disease was noted to be associated with elevation, i.e., bronchopneumonia. In contrast, Houck and Berman reported that no significant difference in serum ribonuclease activity existed in a variety of individuals having carcinoma as compared to a normal group. In that study, 36 normals were compared to 56 patients with carcinoma, sarcoma, thyroid and liver diseases, pancreatitis, and pancreatectomy.ll Three disease states were found to be associated with an elevated serum ribonuclease level: congestive heart failure, myocardial infarction, and primary kidney disease with uremia. In 1960,Levy and Rottino tested three groups for their serum ribonuclease activity: 11 normals, 87 cancer patients (Hodgkin's disease, carcinoma, leukemia, and lymphosarcoma), and 30 noncancer patients (cirrhosis, fractures, and stab wounds).' They listed the patients in each group according to their level of serum ribonuclease activity: high, normal, or low (Table 1). The highest serum ribonuclease activity was found in those patients with cirrhosis (71Vo), while 62q0of the patients with leukemia, 29% of the patientg with carcinoma, and 30% of the patients with Hodgkin's disease demonstrated high serum ribonuclease activity. In 1962, Zytko and Canter0 measured the serum ribonuclease activity in 50 healthy indi-
viduals, 18 untreated cancer patients, and 22 treated cancer paticnts.l The mean serum ribonuclease activity in the untreated cancer group was found to be 87Vo higher than that observed in the normal individuals (Table 2). In addition, a 90% increase in ribonuclease activity was noted in 22 treated cancer patients in whom the disease was considered to be arrested. These authors suggested that the finding might indicate that residual disease was still present and a latent pathological state existed which could not be defined by routine examination and diagnostic procedure. In 1969, Hisada evaluated the serum ribonuclease activity in 55 healthy individuals, 55 cancer patients, and 31 patients having nonneoplastic diseases.6 Elevated serum ribonuclease activity was noted in patients having cancer of the gall bladder and bile duct as well as in patients with either pancreatic, colon, or rectal cancers. In 1973, Chretien, Matthews, and Twomey determined the serum ribonuclease activity in 49 normal individuals as well as 94 patients with cIinically operable malignancies.' Mean serum ribonuclease activities were highest in 22 patients with adenocarcinoma (p C 0.003) and 28 patients with squamous cell carcinoma (p < 0.01)as compared to controls. However, they noted that the mean serum ribonuclease activity in 21 patients with sarcoma and 23 patients with melanoma having either localized or metastatic disease did not differ significantly from controls (Tables 3 and 4). It should be indicated that the studies delineated above (Tables 1 through 4) were based on serum ribonuclease activity as measured on the basis of acid-soluble degradation of the substrate yeast ribonucleic acid.I9 With the availability of synthetic polynucleotides, it has become evident that their use as a substrate within the current assay has significant advantage, i.e., the substrate does not require purification prior to u s e and allows for higher reproducibility because of its homogeneous composition.r*ro-uThe following studies all utilized synthetic polynucleotides as substrates within the assay. Fink, Adam, and Skoog' tested the serum ribonuclease activity in 53 untreated multiple myeloma patients and compared this activity to 31 normals. Of the multiple myeloma patients, 93Vo were demonstrated to possess significantly
January I979
TABLE 1 RibonuclaK in Saum of Hurrmn
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Number
Number of patients Normal Hodgkin’s disase Carcinoma Leukemia Lymphowcoma Cirrhosis Rheumaticheart disease Acute glomaukr nephritis Acute pancreatitis Dmbetic ulcer Stab WOUD~S and lacerations Fractures E!mphugd bleedin# Intestinal obstructions
Normal
11
0.180-0.240
56 21 9 1 14 2
0.129-0.346 0.182-0.355 0.124-0.340 0.461 0.229-0.538
1
4
0.354 0.148 0.290 0.179-0.269
5 2 2
0.258-0.286 0.2760.311 0.232-0.486
1
1
0.250-0.264
High 16 6
5 1 10
2 1
1 2 5
2 1
From Levy. A. L. and Rottino, A.. Clh. Chem. (Winston-Salem,N.C.). 6.43.1960. With permission.
TABLE 2 Serum Ribonudaw Led in 18 Untrated Cmcer Padenh and in 22 Treated cancer Patknts
Patient no.
1 2 3 4 5 6 7 8
9 10 11 12
13 14 I5 16 17 18
Diagnosis
carcinoma of the uterus Carcinoma of the uterus Carcinoma of the uterus carcinomaof the uterus Carcinoma of the utrms Carcinoma of the uterus Carcinoma of the uterus Carcinoma of the utMu Carcinoma of the uterus Carcinoma of the uterus Carcinoma of the breast Astroeytoma of the rectum Carcinoma of the bronchus and skin Carcinoma of the rectum Lymphoma of the B r i l l - s ~ m type ~~~r~ Carcinoma of the lip Carcinomaof the submaxillarygland Carcinoma of the bronchus MC8ll
19
Cueinomaof the uterus
Specific activity
Treatment
40 38 37 33 28 26 24 23 22 20
24 100
59 27 27 26 23 22
30 f 9.7
311
Radiotherapy
91
92
CRC Criricd Reviews in Clinical Luborotory Sciences TABLE 2 (continued) Saum RIbonuclwc Levd in 18 Untreated Cancer Patients and in 22 Treated CMer Patients
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Patient no.
Diagnosis
Specific activity
m
Carcinoma of the uterus
37
21
Carcinoma of the UICNS
28
22 23
Carcinoma of the breast Carcinoma of the breast
42 36
24
Carcinoma of the breast
35
2s
Carcinoma of the breast
31
26
Carcinoma of the breast
27
27 28 29
Carcinoma of the breast Carcinoma of the breast Carcinoma of the prostate
27 26
30 31
Lipomcoma
B u d carcinoma
12 37
32
Carcinoma of the nowpharynx
31
33
Hodgkin’s disease
32
34
Carcinoma of the lung
28
35 36 37
Carcinoma of the skin Carcinoma of the skin Carcinoma of the lip
21 27 24
38 39
Carcinoma of the bronchus Fibroblastic sarcoma
20
40
Carcinoma of the thyroid
I5
50
17
Treatment
Radiotherapy Cobalt therapy Surgery Radiothe ra py Radiothe ra py Radiotherapy Radiothe ra py S~OCry
Surgery Radiotherapy Surgery Radiotherapy Surgery, radiotherapy Radiothe ra py Radiotherapy Surgery suroery Surgery. radiothe ra py Surguy Radiotherapy Surgery
31 t 8.4
MCOll
16 f 2.5
Modified from Zytko. J. and Canttro. A,. Can. Med. Aspa: J.. 86, 482. 1962. With permission.
January 1979
93
TABLE 3 Saum RlbonudaaeLev& in Normal Controls m d Piticnu with cliniully Operable M-&
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Group
Number studied
Normal controls Admocorcinomaa Squamous carcinomas Sarcomaa Melmomcu
* *
49 22 28
21 23
Mean OD'
SEM'
0.259 0.320 0.300 0.279 0.272
0.0095 0.0145 0.01 18 0.0133 0.0139
95% Confidence limits of meno 0.240-0.278 0.290-4.349 0.2754.326 0.2524.303 0.U3-0.302
Statistical difference from normals'
p C 0.003 p C 0.01
Not significant
Not significant
Optical density units per I ml of serum. Standard error of the m a n . By standard error of the difference between means.
From Chrctien, P. B.. Matthews, W., and Toomey. P. L.,Cancer(Philadclphia). 31,175,1973.With permission. TABLE 4 A Complriroa of Tumor Stages and Serum Ribonuclesre Lev&
Number of patients Adenocarcinomas LoCal Metastatic Squamous carcinomas Local Metastatic
'
14 8 28 13
Mean O D
0.318
0.0144 0.0146
0.298 0.301
0.0120 0.01 17
0.321
I5
SEM'
Optical density units per I ml of serum. Standard error of the mean.
From Cluetien. P. B., Matthews, W.. and Toomey. P. L., Cancer (Philadelphia), 31, 175, 1973. With permission.
(p < 0.001) elevated alkaline serum ribonucleast activity (with poly C utilized as substrate). They studied the responsiveness of six patients with multiple myeloma, who were treated with cyclophosphamide and melphalan and noted that the four who responded had developed a decreaselin serum ribonuclease activity, while the two nonresponsive patients showed a progressive increase in the level of Serum ribonuclease activity. Fink and cob leagues also noted that very high serum ribonuclease levels were found in cases of renal disease of varied etiology such as chronic pyelonephritis, polycystic kidney, hydronephrosis, arterionephrosclerosis cystinuria, Fanconi syndrome, and carcinoma of the kidney. More moderate elevations were seen in some
cases of glomerulonephritis and disseminated lupus erythematosus.' Reddi and Holland'o studied the level of ribonuclease activity in patients with carcinoma of the pancreas and compared such activity to that found in normal controls as well as patients with pancreatitis and several other carcinomas (Table 5). Serum ribonuclease activity was based on the degradation of poly C at pH 6.5.1° These authors found that marked elevation of acidic ribonuclease activity occurred in 90% of the patients with carcinoma of the pancreas who possessed normal values of creatinine. Elevation of ribonuclease activity was also noted in six patients with renal failure who had elevated creatinine Ievels. Patients with pancreatitis were noted to have normal values.
CRC Critical Reviews in Clinical Luboratoty Sciences
94
TABLE 5
Serum Ribonuclew of Nonnd Persona and Patients witb Pancrcatitis and Varied NCOPIM~S Number of patients
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Donor Normal Pancrcatitis Pancreatic cancer Miscellaneous cancer Breast Colon Lung Stomach Liver Kidney cancer‘
52 10 30 69 28 20
I1 6 4 4
Average ribonuclease units per ml of serum
Standard deviation
104 120 383 164 131 197 173 194 152 628
24.3 39.4 145.2 58.9 49.1 53.2 57.5 61.4 44.6 89.4
2 SD 48 80 290 120
-
Standard error 3.4 12.5 26.5 7. I 9.3 11.9 17.3 25.1 22.3 44.1
Probability that normal equals 0.17
lo-” 10-6
10-
-
Note: Reaction mixtures consisted of 0.05 ml of poly C ( I 0 0 ug), 0. I5 ml of phosphate and borate buffer (0.1 Mwith respect to phosphate). pH 6.5, and 0.05 ml of serum that had been diluted 200-fold were incubated at 37OC for 15 min and then transferred to an ice bath. To each tube was added and mixed 0.25 ml of cold I .2 M HCIO. containing 0.02 M lanthanum nitrate. After 20 min at 0”. the precipitates were removed by centrifugation at 12.100 x g for 30 min in the cold. The supernatants were diluted with H20and their absorbances were measured at 278 nm. Enzyme and substrate blanks were always run side by side.
’
Probability that miscellaneous cancer equals pancreatic cancer. Serum cratinine values were elevated in each of thebe patients, suggesting false positive values.
From Reddi. K.
K. and Holland, J. F.. Proc. Natl. Aced. Sci. U.S.A.,73,2308.
Reddi and Holland noted that two patients with carcinoma of the gall bladder, two patients with ampullary carcinoma, and one patient with ovarian carcinoma all possessed elevated serum ribonuclease activity, had upper abdominal masses, and were jaundiced. Elevated activity was also found in breast cancer (lo%), colon cancer (1 5%), and lung cancer ( 1 8%). The serum ribonuclease activity of 385 cancer patients was compared to that of 168 normal controls at the Baltimore Cancer Research Center over the last several In these studies, ribonuclease activity was determined at pH 7.5 and three separate substrates were utilized (poly C , poly U,and poly Aspoly U).The mean serum ribonuclease activity the standard deviation for the normal group was 36 f 9.8 units per milliliter against poly U,850 f 248 units per miILiliter against poly C, and 3.9 f 0.8 units per milliliter asainst poly A’poly U. The upper limit of “normal” for serum ribonuclease activity was fixed at two standard deviations above the mean for healthy donors (55.6 units per milliliter vs. poly U, 1346 units per milliliter vs. poly C, and 5.S units per milliliter vs. poly A-poly U). Eleven types of malignan-
*
1976. With permission.
cies were represented in the cancer group: acute nonlymphocytic leukemia (ANLL), brain tumor, carcinoma of the breast, cervical carcinoma, chronic myelocytic leukemia, colorectal carcinoma, Hodgkin’s disease (HD),carcinoma of the lung, non-Hodgkin’s lymphoma (NHL), pancreatic carcinoma, and sarcoma. A summary of the results of these studies is depicted in Table 6. Analysis revealed a significant difference in mean activity between the normal group and each individual cancer group for all of the substrates studied (p
89
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
ALTERATION OF HUMAN SERUM RIBONUCLEASE ACTIVITY IN MALIGNANCY
Authors: Dalia Maor. Michael R. Mardiney, Jr.,*+ Section of Immunology and Cell Biology Baltimore Cancer Research Center Baltimore, Maryland
Referee:
Paul 9. Chrctien Tumor Immunology Section Suriay Branch National Cancer Institute Bethcsda. Maryland
INTRODUCTION The association of elevation of alkaline Serum ribonuclease activity and cancer noted by several investigators more than two decades ago'.' was clouded by both the absence of elevated activity in all cancer'." and the occurrence of high levels of enzyme activity in a variety of unrelated. nonneoplastic diseases.'-" These results together with a lack of knowledge concerning the source, nature, and biologic significance of ribonuclease activity lead only to a few systematic attempts to determine the usefulness of this finding in clinical oncology. Since then, improved methods for the analysis of serum ribonuclease activity utilizing a battery of synthetic polynucleotides as substrates instead of RNA or a single substrate have yielded greater reproducibility and made possible the detection of subtle changes in enzyme activity not previously noted. This review will summarize those observations which demonstrate the association of alteration of ribonuclease activity in malignancy and will suggest that multiple patterns of change in ribonuclease activity exist within the cancer state. These consist mainly of the elevation of ribonuclease ac-
EVIDENCE DEMONSTRATING THE ASSOCIATION BETWEEN INCREASED SERUM RIBONUCLEASE ACTIVITY AND CANCER Most studies concerning the alteration of serum ribonuclease activity in cancer patients have compared either enzyme activity among normal cancer patients and patients with nonmalignant diseases-the level of serum ribonuclease activity pre- and post-cancer therapy which included either surgery, irradiation, or chemotherapy done or in combination or the level of serum ribonuclease activity among patients in remission, relapse, or those considered cured (nonrecurrence for 5 years or more posttreatment). In 1955 Aleksandrowicz presented the first evidence that there were variations in ribonuclease activity in the urine of leukemia patients.' He noted in further studies that the
Currrrrt add-: 2 Rav-Ashi Street. Rmet-An'v, Israel. Cumnt addnu: How& County Docton' Bldg., 9380Baltimore Natl. Pike. Ellicott City, Md.21043. ~
**
tivity in the serum as well as nonneoplastic tissue and the depression of ribonuclease activity in the tumor cell itself. A possible explanation of these observations will evolve during this review.
~~
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
90
CRC Critical Reviews in Clinical Laboratory Sciences
serum ribonuclease activity of patients with chronic granulocytic leukemia was several times higher than that noted in normal individual^.*^1' Also in 1955, Metais tested ribonuclease activity in the serum of 60 young normal males and females, 20 patients with a variety of diseases (tuberculosis, chronic pancreatitis, liver disease, heart disease, and kidney disease), and 16 hospitalized patients with neoplasia.* He found that three patients with pancreatic carcinoma had elevated serum ribonuclease activity while all others studied were in the normal range. In 1958,two groups of investigators reported on alterations in serum ribonuclease activity in cancer. Migliarese tested 75 normals, 48 untreated cancer patients, 34 patients undergoing therapy for carcinoma, and 26 patients with nonmalignant d i ~ e a s e Serum .~ ribonuclease activity was significantly elevated in the untreated cancer patients and either low or within the normal range in patients treated with irradiation or chemotherapy. Only one nonneoplastic disease was noted to be associated with elevation, i.e., bronchopneumonia. In contrast, Houck and Berman reported that no significant difference in serum ribonuclease activity existed in a variety of individuals having carcinoma as compared to a normal group. In that study, 36 normals were compared to 56 patients with carcinoma, sarcoma, thyroid and liver diseases, pancreatitis, and pancreatectomy.ll Three disease states were found to be associated with an elevated serum ribonuclease level: congestive heart failure, myocardial infarction, and primary kidney disease with uremia. In 1960,Levy and Rottino tested three groups for their serum ribonuclease activity: 11 normals, 87 cancer patients (Hodgkin's disease, carcinoma, leukemia, and lymphosarcoma), and 30 noncancer patients (cirrhosis, fractures, and stab wounds).' They listed the patients in each group according to their level of serum ribonuclease activity: high, normal, or low (Table 1). The highest serum ribonuclease activity was found in those patients with cirrhosis (71Vo), while 62q0of the patients with leukemia, 29% of the patientg with carcinoma, and 30% of the patients with Hodgkin's disease demonstrated high serum ribonuclease activity. In 1962, Zytko and Canter0 measured the serum ribonuclease activity in 50 healthy indi-
viduals, 18 untreated cancer patients, and 22 treated cancer paticnts.l The mean serum ribonuclease activity in the untreated cancer group was found to be 87Vo higher than that observed in the normal individuals (Table 2). In addition, a 90% increase in ribonuclease activity was noted in 22 treated cancer patients in whom the disease was considered to be arrested. These authors suggested that the finding might indicate that residual disease was still present and a latent pathological state existed which could not be defined by routine examination and diagnostic procedure. In 1969, Hisada evaluated the serum ribonuclease activity in 55 healthy individuals, 55 cancer patients, and 31 patients having nonneoplastic diseases.6 Elevated serum ribonuclease activity was noted in patients having cancer of the gall bladder and bile duct as well as in patients with either pancreatic, colon, or rectal cancers. In 1973, Chretien, Matthews, and Twomey determined the serum ribonuclease activity in 49 normal individuals as well as 94 patients with cIinically operable malignancies.' Mean serum ribonuclease activities were highest in 22 patients with adenocarcinoma (p C 0.003) and 28 patients with squamous cell carcinoma (p < 0.01)as compared to controls. However, they noted that the mean serum ribonuclease activity in 21 patients with sarcoma and 23 patients with melanoma having either localized or metastatic disease did not differ significantly from controls (Tables 3 and 4). It should be indicated that the studies delineated above (Tables 1 through 4) were based on serum ribonuclease activity as measured on the basis of acid-soluble degradation of the substrate yeast ribonucleic acid.I9 With the availability of synthetic polynucleotides, it has become evident that their use as a substrate within the current assay has significant advantage, i.e., the substrate does not require purification prior to u s e and allows for higher reproducibility because of its homogeneous composition.r*ro-uThe following studies all utilized synthetic polynucleotides as substrates within the assay. Fink, Adam, and Skoog' tested the serum ribonuclease activity in 53 untreated multiple myeloma patients and compared this activity to 31 normals. Of the multiple myeloma patients, 93Vo were demonstrated to possess significantly
January I979
TABLE 1 RibonuclaK in Saum of Hurrmn
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Number
Number of patients Normal Hodgkin’s disase Carcinoma Leukemia Lymphowcoma Cirrhosis Rheumaticheart disease Acute glomaukr nephritis Acute pancreatitis Dmbetic ulcer Stab WOUD~S and lacerations Fractures E!mphugd bleedin# Intestinal obstructions
Normal
11
0.180-0.240
56 21 9 1 14 2
0.129-0.346 0.182-0.355 0.124-0.340 0.461 0.229-0.538
1
4
0.354 0.148 0.290 0.179-0.269
5 2 2
0.258-0.286 0.2760.311 0.232-0.486
1
1
0.250-0.264
High 16 6
5 1 10
2 1
1 2 5
2 1
From Levy. A. L. and Rottino, A.. Clh. Chem. (Winston-Salem,N.C.). 6.43.1960. With permission.
TABLE 2 Serum Ribonudaw Led in 18 Untrated Cmcer Padenh and in 22 Treated cancer Patknts
Patient no.
1 2 3 4 5 6 7 8
9 10 11 12
13 14 I5 16 17 18
Diagnosis
carcinoma of the uterus Carcinoma of the uterus Carcinoma of the uterus carcinomaof the uterus Carcinoma of the utrms Carcinoma of the uterus Carcinoma of the uterus Carcinoma of the utMu Carcinoma of the uterus Carcinoma of the uterus Carcinoma of the breast Astroeytoma of the rectum Carcinoma of the bronchus and skin Carcinoma of the rectum Lymphoma of the B r i l l - s ~ m type ~~~r~ Carcinoma of the lip Carcinomaof the submaxillarygland Carcinoma of the bronchus MC8ll
19
Cueinomaof the uterus
Specific activity
Treatment
40 38 37 33 28 26 24 23 22 20
24 100
59 27 27 26 23 22
30 f 9.7
311
Radiotherapy
91
92
CRC Criricd Reviews in Clinical Luborotory Sciences TABLE 2 (continued) Saum RIbonuclwc Levd in 18 Untreated Cancer Patients and in 22 Treated CMer Patients
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Patient no.
Diagnosis
Specific activity
m
Carcinoma of the uterus
37
21
Carcinoma of the UICNS
28
22 23
Carcinoma of the breast Carcinoma of the breast
42 36
24
Carcinoma of the breast
35
2s
Carcinoma of the breast
31
26
Carcinoma of the breast
27
27 28 29
Carcinoma of the breast Carcinoma of the breast Carcinoma of the prostate
27 26
30 31
Lipomcoma
B u d carcinoma
12 37
32
Carcinoma of the nowpharynx
31
33
Hodgkin’s disease
32
34
Carcinoma of the lung
28
35 36 37
Carcinoma of the skin Carcinoma of the skin Carcinoma of the lip
21 27 24
38 39
Carcinoma of the bronchus Fibroblastic sarcoma
20
40
Carcinoma of the thyroid
I5
50
17
Treatment
Radiotherapy Cobalt therapy Surgery Radiothe ra py Radiothe ra py Radiotherapy Radiothe ra py S~OCry
Surgery Radiotherapy Surgery Radiotherapy Surgery, radiotherapy Radiothe ra py Radiotherapy Surgery suroery Surgery. radiothe ra py Surguy Radiotherapy Surgery
31 t 8.4
MCOll
16 f 2.5
Modified from Zytko. J. and Canttro. A,. Can. Med. Aspa: J.. 86, 482. 1962. With permission.
January 1979
93
TABLE 3 Saum RlbonudaaeLev& in Normal Controls m d Piticnu with cliniully Operable M-&
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Group
Number studied
Normal controls Admocorcinomaa Squamous carcinomas Sarcomaa Melmomcu
* *
49 22 28
21 23
Mean OD'
SEM'
0.259 0.320 0.300 0.279 0.272
0.0095 0.0145 0.01 18 0.0133 0.0139
95% Confidence limits of meno 0.240-0.278 0.290-4.349 0.2754.326 0.2524.303 0.U3-0.302
Statistical difference from normals'
p C 0.003 p C 0.01
Not significant
Not significant
Optical density units per I ml of serum. Standard error of the m a n . By standard error of the difference between means.
From Chrctien, P. B.. Matthews, W., and Toomey. P. L.,Cancer(Philadclphia). 31,175,1973.With permission. TABLE 4 A Complriroa of Tumor Stages and Serum Ribonuclesre Lev&
Number of patients Adenocarcinomas LoCal Metastatic Squamous carcinomas Local Metastatic
'
14 8 28 13
Mean O D
0.318
0.0144 0.0146
0.298 0.301
0.0120 0.01 17
0.321
I5
SEM'
Optical density units per I ml of serum. Standard error of the mean.
From Cluetien. P. B., Matthews, W.. and Toomey. P. L., Cancer (Philadelphia), 31, 175, 1973. With permission.
(p < 0.001) elevated alkaline serum ribonucleast activity (with poly C utilized as substrate). They studied the responsiveness of six patients with multiple myeloma, who were treated with cyclophosphamide and melphalan and noted that the four who responded had developed a decreaselin serum ribonuclease activity, while the two nonresponsive patients showed a progressive increase in the level of Serum ribonuclease activity. Fink and cob leagues also noted that very high serum ribonuclease levels were found in cases of renal disease of varied etiology such as chronic pyelonephritis, polycystic kidney, hydronephrosis, arterionephrosclerosis cystinuria, Fanconi syndrome, and carcinoma of the kidney. More moderate elevations were seen in some
cases of glomerulonephritis and disseminated lupus erythematosus.' Reddi and Holland'o studied the level of ribonuclease activity in patients with carcinoma of the pancreas and compared such activity to that found in normal controls as well as patients with pancreatitis and several other carcinomas (Table 5). Serum ribonuclease activity was based on the degradation of poly C at pH 6.5.1° These authors found that marked elevation of acidic ribonuclease activity occurred in 90% of the patients with carcinoma of the pancreas who possessed normal values of creatinine. Elevation of ribonuclease activity was also noted in six patients with renal failure who had elevated creatinine Ievels. Patients with pancreatitis were noted to have normal values.
CRC Critical Reviews in Clinical Luboratoty Sciences
94
TABLE 5
Serum Ribonuclew of Nonnd Persona and Patients witb Pancrcatitis and Varied NCOPIM~S Number of patients
Critical Reviews in Clinical Laboratory Sciences Downloaded from informahealthcare.com by McMaster University on 11/19/14 For personal use only.
Donor Normal Pancrcatitis Pancreatic cancer Miscellaneous cancer Breast Colon Lung Stomach Liver Kidney cancer‘
52 10 30 69 28 20
I1 6 4 4
Average ribonuclease units per ml of serum
Standard deviation
104 120 383 164 131 197 173 194 152 628
24.3 39.4 145.2 58.9 49.1 53.2 57.5 61.4 44.6 89.4
2 SD 48 80 290 120
-
Standard error 3.4 12.5 26.5 7. I 9.3 11.9 17.3 25.1 22.3 44.1
Probability that normal equals 0.17
lo-” 10-6
10-
-
Note: Reaction mixtures consisted of 0.05 ml of poly C ( I 0 0 ug), 0. I5 ml of phosphate and borate buffer (0.1 Mwith respect to phosphate). pH 6.5, and 0.05 ml of serum that had been diluted 200-fold were incubated at 37OC for 15 min and then transferred to an ice bath. To each tube was added and mixed 0.25 ml of cold I .2 M HCIO. containing 0.02 M lanthanum nitrate. After 20 min at 0”. the precipitates were removed by centrifugation at 12.100 x g for 30 min in the cold. The supernatants were diluted with H20and their absorbances were measured at 278 nm. Enzyme and substrate blanks were always run side by side.
’
Probability that miscellaneous cancer equals pancreatic cancer. Serum cratinine values were elevated in each of thebe patients, suggesting false positive values.
From Reddi. K.
K. and Holland, J. F.. Proc. Natl. Aced. Sci. U.S.A.,73,2308.
Reddi and Holland noted that two patients with carcinoma of the gall bladder, two patients with ampullary carcinoma, and one patient with ovarian carcinoma all possessed elevated serum ribonuclease activity, had upper abdominal masses, and were jaundiced. Elevated activity was also found in breast cancer (lo%), colon cancer (1 5%), and lung cancer ( 1 8%). The serum ribonuclease activity of 385 cancer patients was compared to that of 168 normal controls at the Baltimore Cancer Research Center over the last several In these studies, ribonuclease activity was determined at pH 7.5 and three separate substrates were utilized (poly C , poly U,and poly Aspoly U).The mean serum ribonuclease activity the standard deviation for the normal group was 36 f 9.8 units per milliliter against poly U,850 f 248 units per miILiliter against poly C, and 3.9 f 0.8 units per milliliter asainst poly A’poly U. The upper limit of “normal” for serum ribonuclease activity was fixed at two standard deviations above the mean for healthy donors (55.6 units per milliliter vs. poly U, 1346 units per milliliter vs. poly C, and 5.S units per milliliter vs. poly A-poly U). Eleven types of malignan-
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1976. With permission.
cies were represented in the cancer group: acute nonlymphocytic leukemia (ANLL), brain tumor, carcinoma of the breast, cervical carcinoma, chronic myelocytic leukemia, colorectal carcinoma, Hodgkin’s disease (HD),carcinoma of the lung, non-Hodgkin’s lymphoma (NHL), pancreatic carcinoma, and sarcoma. A summary of the results of these studies is depicted in Table 6. Analysis revealed a significant difference in mean activity between the normal group and each individual cancer group for all of the substrates studied (p
