Hemostatic Abnormalities in Malignancy, a Prospective Study of One Hundred Eight Patients Part I. Coagulation Studies NORA C. J. SUN, M.D., WILLIAM M. McAFEE, M.Sc, GILBERT J. HUM, M.D., AND JOHN M. WEINER, D.P.H. Sun, Nora C. J., McAfee, William M., Hum, Gilbert J., and Weiner, John M.: Hemostatic abnormalities in malignancy, a prospective study of one hundred eight patients. Part I. Coagulation studies. Am J Clin Pathol 71: 10-16,1979. A prospective study of hemostatic abnormalities in 108 cancer patients was undertaken at an oncology clinic in a university teaching hospital. Tests included Quick prothrombin time, activated partial thromboplastin time, thrombin time, platelet count, modified Ivy bleeding time, fibrinogen, fibrin degradation products (FDP), euglobulin lysis time, protamine sulfate test, and factor V, VII, VIII and X assays. Ninety-eight per cent of the patients had one or more abnormal coagulation tests. The commonest abnormalities were elevated fibrin degradation products and prolonged thrombin time. Thrombocytosis occurred in 57% of patients, hyperfibrinogenemia in 46%, thrombocytopenia in 11%, and none had hypofibrinogenemia. It is suggested that platelet count, fibrinogen concentration, and serum FDP assay are the most useful tests in assessing the hemostatic abnormalities in cancer patients, although thrombin time, factor V assay, and bleeding time may also be helpful. The peripheral blood smears of 53 patients were reviewed, and only one showed microangiopathic hemolytic anemia. The data illustrate that subclinical coagulopathy is relatively frequent in patients with malignancy. (Key words: Disseminated intravascular coagulation; Cancer; Hypercoagulable state; Microangiopathic hemolytic anemia.) Departments of Pathology and Internal Medicine, USC-John Wesley Hospital, Los Angeles, California of clotting abnormalities in cancer patients. 36 This study was designed to evaluate coagulation prospectively in a series of consecutive patients referred to an oncology clinic for staging and consideration of chemotherapy for their malignant disease. The purpose was to examine the incidence and types of hemostatic derangement in these patients by conventional methods. The concept of "intravascular coagulation and fibrinolysis syndrome" (ICF) of Owen and Bowie26 was adopted, and their classification of "overcompensated, compensated and decompensated I C F " was tested. Materials and Methods One hundred eight consecutive new patients referred to the oncology service at the University of Southern California-John Wesley County Hospital during a period of one year with tissue-proven malignancy who had relatively complete clinical and laboratory data were included and formed the basis for the study. As part of the initial work-up, coagulation studies included Quick prothrombin time (FT), activated partial thromboplastin time (APTT), thrombin time (TT), platelet count (by thrombocounter), modified Ivy bleeding time, serum fibrin degradation products (FDP), 12 protamine sulfate test, 17 euglobulin lysis time, 2 fibrinogen concentration (biuret method), 2 and factor V, VII, VIII, and X assays. 2 The study group consisted of 54 men and 54 women. Ages ranged from 16 to 86 years for men (median 61) and from 22 to 86 years for the women (median 56). The combined median was 59 years, and the mean was 53 years. More than half the patients (68) were in the sixth and seventh decade of life. The primary sites of malignant disease are listed in Table 1. Four patients had a second primary tumor. These included tumors of the breast (2), pancreas (1), THE ASSOCIATION of recurrent thrombotic episodes with malignant disease was initially described by Trousseau a century ago, 38 and migratory thrombophlebitis has been suggested as the first clinical indication of an occult malignancy. 10 ' 40 Isolated reports of bleeding diathesis and/or thrombosis occurring in patients with malignancy have been interpreted as secondary to disseminated intravascular coagulation (DIC) or the defibrination syndrome. 5,9 ' 30 A previous study by one of us (N.C.J.S.) based on cases referred for consultation to a coagulation laboratory disclosed a high incidence Received November 8, 1977; received revised manuscript and accepted for publication January 9, 1978. Supported in part by John Wesley Attending Staff Account #6002. Presented in part at the American Society of Clinical Pathologists Meeting, October 27, 1976, Los Angeles, California (abstract, Am J Clin Pathol 67:206, 1977). Address reprint requests to Dr. Sun: Department of Pathology, Box 22, Harbor General Hospital, 1000 W. Carson Street, Torrance, California 90509. 0002-9173/79/0100/0010 $00.85 © American Society of Clinical Pathologists 10 HEMATOSTATIC ABNORMALITIES IN MALIGNANCY Vol. 71 • No. I and stomach (1). The most frequent histologic types were adenocarcinoma (53 patients), squamous-cell carcinoma (20), large-cell anaplastic carcinoma (7), myeloproliferative disorders (5), and undifferentiated smallcell carcinoma (3). Other types of solid tumor included malignant melanoma, glioblastoma multiforme, osteogenic sarcoma, Ewing's sarcoma, embryonal carcinoma, and non-Hodgkin's lymphoma. The extent of tumor-cell differentiation was classified according to Broders' description.3 Eighty-seven patients (81%) had metastatic disease determined by one or more of the following: physical examination, roentgenography of the chest, liver scan, bone scan, skeletal survey, bone marrow aspiration and/or biopsy, or other tissue diagnosis. Nineteen patients (18%) were found to be free of evident metastatic disease. Fifty-seven patients had two or more sites of metastases. The lung was the most common location of metastatic disease (41 patients), followed by lymph nodes (38), bone (37), soft tissue (30), liver (27), and brain (8). About half the patients (51) had histories related to their malignant disease of less than four months, and 71% of the patients had had the disease for less than 12 months. Other associated clinical conditions included a recent history of thrombophlebitis (3 patients), pulmonary embolism (2) and gram-negative septicemia (1). Six patients had slight concurrent bleeding, which was related to their malignant diseases. These were menorrhagia associated with acute myelogenous leukemia (1), gastrointestinal bleeding from carcinoma of the colon (2), bleeding from carcinoma of the stomach (2), and bleeding from cancer of the oral cavity (1). Since clotting and fibrinolytic systems are in a constant dynamic balance, Owen and Bowie suggested the term "intravascular coagulation and fibrinolysis syndrome" (ICF) to emphasize this relationship. Experimentally, they and others613-20-25 observed several 11 Table 1. Primary Sites of Malignant Disease Sites Number Lung Breast Unknown primary site Colon Bone marrow Prostate Ovary, brain, skin, esophagus Kidney, testicle, stomach, oral cavity Liver, lymph node, middle ear, thyroid, tongue, and bone 27 21 12 11 7 4 3 each 2 each 1 each patterns of this syndrome, which were dependent on the triggering mechanism, the dose and duration of stimulus, and the compensatory abilities of bone marrow and liver. It is generally accepted that an elevation of FDP usually indicates plasminic digestion of fibrinogen and/or fibrin; and it is, in most circumstances, suggestive of an ICF syndrome.14 Supplemented by other hemostatic tests, such as platelet count or fibrinogen level, Cooper and associates7 classified ICF into three groups: decompensated, which is defined by depressed fibrinogen or platelet levels along with other hemostatic abnormalities; compensated, in which fibrinogen or platelet values are normal, but other hemostatic tests are abnormally prolonged; overcompensated, where the platelets or fibrinogen are increased, together with other hemostatic abnormalities. This has also been correlated with the conventional terms: acute, subacute, and chronic DIC with decompensated, overcompensated, and compensated ICF, respectively.1-29-33 We applied this classification to our study, and a comparative evaluation of clinical conditions and laboratory tests was made. Results One hundred six of the 108 patients (98%) had one or more abnormal coagulation test results. Two patients Table 2. Results of Coagulation Studies Low Test Platelets Bleeding time APTT Quick PT Fibrinogen FDP Protamine sulfate test Euglobulin lysis time Factor VIII Factor VII Factor X Factor V TT Normal Range 170,000-300,000/mm < 6 min 30-43 sec 11-14 sec 0.18-0.45 g/dl <10 j/.g/ml Negative >2hr 60-140% 80-155% 75-135% 88-138% Control ± 2 sec No. 3 Normal % No. % No. % 34 74 74 91 58 35 97 93 44 43 54 33 33 32 73 69 84 54 32 90 87 44 43 54 33 31 62 28 25 15 50 73 11 57 27 23 14 46 68 10 — — 23 23 12 14 75 23 23 12 14 69 12 11 — — — — — — 9 2 0 14 33 34 34 53 0 High 8 2 0 13 33 34 34 53 0 SUN ET AL. 12 A.J.C.P. . January 1979 Table 3. Pearson Correlation Coefficient (T) among Pairs of Laboratory Results Erythrocytes Leukocytes Platelets Bleeding time APTT PT Fibrinogen FDP PSTt ELTt Factor VIII Factor VII Factor X Factor V ATT .21* .04 .18* .16 -.05 -.00 .04 .02 .10 .01 .07 -.17* -.00 -.08 .46* .11 -.20* -.08 -.09 .20* .14 .03 -.09 .33* -.08 -.05 .07 -.08 -.13 -.13 .09 .12 .23* -.09 .00 -.03 -.07 .00 .07 .05 .13 Platelets .06 .21* -.22* .02 Bleeding time -.00 .32* .03 -.18* .16 -.18* .01 -.03 -.05 .02 C WBC ErythLeurokocytes cytes -.05 .25* .14 -.02 .07 .12 -.03 .22* .02 -.14 -.03 .11 -.01 .24* .13 -.16 -.16 -.01 .26* APTT -.31* -.26* -.16 .05 PT -.23* -.06 .18* .26* FDP .05 -.03 .01 .09 PSTt .01 .10 .09 .21* Fibrinogen -.13 .13 -.09 -.22* -.14 ELTt -.16 -.11 .17* .21* Factor VIII .28* -.02 -.21* Factor VII .20* -.02 Factor X .25* Factor V ATT t Euglobin lysis time. ATT = difference between patient's TT and control's TT. * P < 0.05. t Protamine sulfate test. had one abnormal test, five had two, 13 had three, 14 had four, 24 had five, 16 had six, 18 had seven, nine had eight, five had nine, and two had ten. Two patients had normal results. One was a 62-year-old man who had a non-Hodgkin's lymphoma, and the other was a 47-year-old woman with mammary carcinoma. Neither had any evidence of metastatic disease. The two patients with ten abnormal clotting tests had bronchogenic carcinoma with widespread metastases. A summary of coagulation laboratory data from these 108 patients is shown in Table 2. Elevated FDP (68%) and prolonged thrombin time (69%) were the most com- mon abnormalities. These two tests also had significant correlation (Table 3). Meaningful correlations were observed among pairs of tests, such as PT with APTT, PT with FDP, PT with factor VII, PT with factor X, factor X with factor VII, fibrinogen with platelets, FDP with APTT, FDP with PT, FDP with factor VIII, FDP with ATT, factor V with ATT. Some of the paired tests were negatively correlated, such as PT with factor VII; i.e., the lower the factor VII activity, the longer the prothrombin time. None of the tests could replace another in spite of the significant relationships, because the correlation coefficients are not near ± 1 in value. Table 4. Pertinent Clinical and Laboratory Patient Patient Patient Patient Patient Patient Patient Patient Patient Patient Patient Patient 2 6 7 24 31 36 65 69 84 90 93 104 * ND = not done. t Bleeding time. Age (Years)/Sex Primary Tumor 52/F 59/F 73/F 52/F 42/M 61/M 64/F 62/F 56/M 76/M 55/F 50/F Acute myelogenous leukemia Breast Lung Breast Stomach Colon Oral cavity Malignant melanoma Preleukemia Acute myelogenous leukemia Breast Myelofibrosis and myeloid metaplasia t Protamine sulfate test. § Euglobulin lysis time. Platelet Count (/mm3) BTt (Min) APTT (Sec) PT (Sec) ATT 97 x 103 124 x 103 44 x 103 106 x 103 167 x 103 150 x 103 124 x 103 167 x 103 124 x 103 44-5.2 158 x 103 6.5 6.6 ND* 4.3 5.5 6.1 6.0 5.5 7.0 5.2 3.7 60 40 35 42 46 33 52 35 40 36 30 15 12 18 13 15 12 14 12 14 12 12 1 1 0 3 2 9 3 4 9 7 3 36 x 103 3.8 26 12 4 13 HEMATOSTATIC ABNORMALITIES IN MALIGNANCY Vol. 71 • No. I Fifty-seven per cent of the patients had thrombocytosis, and 11% had thrombocytopenia. Of the latter, four had platelet counts below 100,000/mnr'1 (Table 4). Three of these four patients had myeloproliferative disorders. The fourth patient was a 73-year-old woman with metastatic bronchogenic carcinoma. There was no relationship between the histologic type and the extent of tumor cell differentiation and elevation of FDP, or between the duration of illness and any of the tests. Peripheral blood smears from 53 of the patients revealed only one instance of microangiopathic hemolytic anemia. This patient had a metastatic mucin-producing adenocarcinoma of the stomach. Normal or elevated platelet counts were frequently seen in those patients receiving radiation therapy. Seven of nine patients recovering from operations (including biopsy) had thrombocytosis (Table 5). All of these patients had clinical or histologic evidence of metastases. Adopting Owen and Bowie's hypothesis, an attempt was made to identify those key tests that might enable us to discover patients with coagulation problems. We used FDP and platelets as the indicators and separated the patients into three groups: the patients with no ICF (normal FDP); those with compensated ICF (elevated FDP, but normal platelets); and those with overcompensated ICF (elevated FDP and platelets). The number of patients who might have a true decompensated ICF (elevated FDP and decreased platelet count) was relatively small (Table 4), and they were included in the compensatory ICF for the statistical purpose. We then compared a battery of hemostatic tests among these three groups (Table 6). Several tests appeared to be helpful in differentiating those with no ICF, compensated ICF, and overcompensated ICF. These were bleeding time, fibrinogen, factor V, and thrombin time. However, no statistically significant difference among the clotting tests was found when we used FDP and fibrinogen, except in the case of the platelet count. Thirty per cent of the patients who had bronchogenic carcinomas showed a compensated ICF state, and 52% overcompensated. About half (48%) of the patients with mammary cancers did not have ICF, while 46% of the patients with colonic carcinoma had overcompensated ICF. Those patients with unknown primary tumors were largely in the overcompensated state (58%), and only 17% had no evidence of ICF. Sixty-seven per cent of the patients who had bronchogenic carcinomas and 66% of those with unknown primary tumors also had hyperfibrinogenemia. The same criteria were applied to different clinical conditions. Patients with thrombophlebitis or pulmonary embolism also had laboratory evidence of overcompensated ICF. However, no definite pattern was seen in the patients with current bleeding, and such bleeding was most likely to be a result of their primary tumors rather than a manifestation of ICF syndrome. As might be expected, patients who had metastases of the lung, brain, liver, and lymph nodes and those with two or more sites of metastases were frequently in the overcompensated ICF category. Discussion Viable malignant cells circulate in the blood or lymphatic systems, or both. 16,31 Experimental studies have demonstrated intravascular cancer localization by the adhesion of cancer cells to the vascular endothelium and the formation of a fibrin thrombus. 35 Laki 18 showed that a proper fibrin network is essential for the development of blood supply for tumors. Acceleration Findings in Patients with Low Platelet Counts Fibrinogen (g/dl) Factor VIII Factor VII Factor X Factor V (%) (%) (%) (%) Bleeding Thrombosis FDP PSTt ELT§ (Hr) _ + + + + + - >2 >2 >2 >2 >2 >2 >2 >2 >2 1.5 >2 100 21 250 25 50 66 ND* 85 300 135 50 86 140 23 80 30 120 ND 120 60 72 160 62 86 46 62 62 100 ND 94 100 82 100 60 52 27 36 74 41 ND 100 120 150 170 Yes No No No Yes Yes Yes No No No No No No No No No No No No No No No >2 ND ND ND ND No No 230 370 180 390 210 460 250 390 510 240 320 + + + — + + - 860 + _ SUNCTAZ.. 14 A.J.C.P. • January 1979 Table 5. Selective Clinical and Laboratory Information about Patients Recently Treated for Their Malignant Disease Type of Treatment Patient's Age (Years)/Sex Radiation therapy Patient 106 Patient 101 55/F 57/F Patient 98 Patient 80 Patient 42* Patient 25 Patient 5 Surgical treatment Patient 12 Patient 19 Patient 42* Patient 45 Patient 48 Patient 69 Patient 79 Patient 82 Patient 94 Fibrinogen (g/dl) Abnormal FDP (>10/xg/ml) 410,000 0.61 Yes 728,000 450,000 380,000 585,000 300,000 345,000 0.61 0.40 0.60 0.64 0.28 0.50 Yes Yes Yes No Yes Yes 443,000 > 1,000,000 585,000 400,000 742,000 167,000 190,000 360,000 774,000 0.38 0.26 0.64 0.19 0.67 0.39 0.19 0.38 0.32 Yes No No Yes Yes No Yes No No Platelets (/mm3) Primary Tumor 56/M 34/F 58/M 50/M 60/M Breast Unknown (large cell undifferentiated) Unknown (clear cell) Lung Lung Lung Lung 59/M 48/M 58/M 34/M 51/F 62/F 67/F 79/M 22/M Colonic Stomach Lung Glioblastoma multiforme Unknown Malignant melanoma, skin Unknown Malignant melanoma, skin Embryonal, testis * Same patient. of platelet and fibrinogen turnover rates, seen in patients with various kinds' of malignancy,15 have been attributed to their increased consumption within the tumor mass or in the process of intravascular coagulation.34 Deposition offibrin within a necrotic tumor mass has been found histologically. Incorporation of fibrin with tumor cells or an intravascular thrombus within the stromal capillaries of tumor nodules also is seen. Intravascular or intralymphatic tumor infiltration is not uncommonly observed in the histologic sections from surgical or autopsy specimens, and is often a sign of early dissemination of the disease and a poor prognosis. Thrombocytosis, hyperfibrinogenemia, and elevation of plasma levels of factors II, V, VII, IX, and X with acceleration of thromboplastin generation have been described.823,28 These findings have been interpreted as a compensatory overproduction secondary to a low-grade intravascular coagulation. Chronic and subacute DIC are probably far more common than the dramatic acute DIC.27 However, the definitive diagnoses of these conditions are rather difficult; they often require kinetic studies of platelets and coagulation factors.22 Such studies are not available in service laboratories. Thirteen routine tests were performed in our study. The platelet count, fibrinogen concentration, serum FDP assay, and thrombin time determination were the tests most important in evaluation of the balance between the clotting and fibrinolytic systems (Table 6). Microangiopathic hemolytic anemia was uncommon in this group of patients. A single abnormal coagulation test may not be significant, because the blood level of any one biological substance reflects the balance between its anabolism and catabolism. However, 88 patients had more than four abnormal test results. This indicates that hemostatic derangement is indeed quite common in cancer patients. The protamine sulfate test was proposed to be very sensitive and specific for intravascular coagulation,,732 but only 11% of our patients had a positive value. A significant correlation betwen protamine sulfate test results and FDP was observed (Table 3). The discrepancy in the incidences of positivity of FDP and protamine sulfate test results might be due to the differences in sensitivity and specificity of the methods or the differences in the clinical stages during the time the patients were studied. The present study indicates that coagulation abnormalities are rather common among cancer patients, even though the incidences of hemorrhage and thrombosis were low. Most patients appeared to be in a delicate hemostatic balance (compensated ICF) or in a hypercoagulable state (overcompensated ICF). An untoward effect might be provoked by stimuli such as surgery, radiation therapy, or infection. Thrombocytosis19 and hyperfibrinogenemia4 in cancer patients comparable to our findings have been reported in other studies. Interestingly, those patients who had hepatic metastasis had normal or high fibrinogen concentrations (14/27 had normal values, and 13/27 had fibrinogen values of more than 0.45 g/dl); 95% of the patients with metastatic disease of the bone had normal or elevated platelet counts (12 normal, two elevated, and two below normal). These results are consistent with the known reserve capacities of both liver and bone Vol. 71 • No. I 15 HEMATOSTATIC ABNORMALITIES IN MALIGNANCY marrow, and they are probably related to the mass of the metastases, the site of primary tumor, and the accuracy of the clinical staging evaluations. Digestion of protein by proteolytic enzymes other than thrombin or plasmin, the redistribution of protein between intra- and extravascular spaces that may occur in diseases, hemoconcentration, and fibrin deposition around cancer cells in the extravascular space may all contribute to the elevation or depression of fibrinogen and platelets. The situation is further complicated by the lack of sensitive tests for differentiating the fibrin from fibrinogen degradation products and the difficulty in differentiating local from disseminated intravascular coagulation and in differentiating intravascular from extravascular coagulation. In our opinion, FDP is the most sensitive test for the evaluation of the ICF syndrome. Its value is enhanced when it is combined with platelet count, fibrinogen concentration, and thrombin time determinations and correlated with clinical and pathologic conditions. We agree with McKay that DIC or ICF is a syndrome and is an intermediary mechanism of the disease.21 In a proper clinical setting, supported by certain abnormal hemostatic test results, this syndrome should be suspected. Furthermore, precautionary measures can be taken to avoid its complications and a grave outcome. Anticoagulants and fibrinolytic agents were utilized in the treatment of experimental malignant tumors, with favorable results.24-39 Clinical trials using warfarin sodium or heparin as an adjunct to conventional chemotherapeutic agents have demonstrated that they might have an enhancing beneficial effect."'37 Nevertheless, more extensive clinical or experimental studies are needed to determine their true value. Table 6. Comparison of Laboratory Data among Different Groups of Patients Test Bleeding time (min) Normal Prolonged APTT (sec) Shortened Normal Prolonged PT (sec) Shortened Normal Prolonged Fibrinogen (g/dl) Low Normal High Protamine sulfate test Negative Positive Euglobulin lysis time (hr) Shortened Normal Factor VIII (%) Low Normal High Factor VII (%) Low Normal High Factor X (%) Low Normal High Factor V (%) Low Normal High Thrombin time (sec) Normal High Normal Range No ICF* (Normal FDP) Compensated ICF (FDPf, Platelets < 300,000/mm3) Overcompensated ICF (FDPj, Platelets > 300,000/mm3) 20 (57%) 15 (43%) 22 (73%) 8 (27%) 32 (87%) 5 (14%) 5 (14%) 26 (74%) 4 (11%) 2 (7%) 20 (65%) 9 (29%) 2 (5%) 28 (67%) 12 (29%) 1 (3%) 32 (91%) 2 (6%) 0 24 (77%) 7 (23%) 1 (2%) 35 (83%) 6 (14%) 0 23 (66%) 12 (34%) 0 21 (68%) 10 (32%) 0 14 (33%) 28 (67%) 34 (97%) 1 (3%) 26 (84%) 5 (16%) 37 (88%) 5 (12%) 5 (14%) 30 (86%) 4 (13%) 27 (87%) 5 (13%) 36 (87%) 12 (36%) 17 (52%) 4 (12%) 11 (41%) 9 (33%) 7 (26%) 10 (25%) 18 (45%) 12 (30%) 5 (15%) 19 (58%) 9 (27%) 10 (37%) 11 (41%) 6 (22%) 19 (48%) 13 (33%) 8 (20%) 12 (36%) 15 (46%) 6 (18%) 9 (33%) 15 (56%) 3 (11%) 13 (33%) 24 (60%) 3 (8%) 24 (73%) 6 (18%) 3 (9%) 17 (63%) 5 (19%) 5 (19%) 12 (30%) 22 (55%) 6 (15%) 18 (51%) 17 (49%) 9 (29%) 22 (71%) 6 (14%) 36 (86%) <6 P < 0.020 30-43 P < 0.231 P < 0.332 11-14 0.18-0.45 P < 0.003 Negative P < 0.184 >2 P < 0.964 60-140 P < 0.271 80-155 P < 0.067 75-135 P < 0.635 88-138 P < 0.014 Control ± 2 * Intravascular coagulation and fibrinolysis syndrome. 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