Document LJDJqkoVxaxQKKzbbEYnYeKnX

TO: FROM: RE: DATE: MEMORANDUM Doug Cohen Study Showing Statistical Correlation between PCBs and Breast Cancer March 25, 1992 Bill Snyder sent me the attached study which shows among other things that mean concentrations of PCBs were 50-60% higher in tissues of women who had breast cancer compared with women in control group who turned out to have non-cancerous growths. Study also indicates that an increase of only 10 parts per billion of PCBs in tissue level corresponds to a 1% increase in risk of breast cancer. This is dynamite. And the defendants still claim PCBs are safe! M A R --2 5 --0 2 M E S I O : 2 e> Eh. . CALL BEFORE SE N SIN G i.k - R-02 Pesticides and Polychlorinated Biphenyl Residues in Hum an Breast Lipids and Their Relation to Breast Cancer FRANK FALK, JfL Department of Ophthalmology University of Michigan Ann Arbor, Michigan ANDREW RICO, Jr. Departments of Pathology and Surgery Hartford Hospital Hartford, Connecticut MARYS. WOLFF JAMES GODBOL Division of Environment*) an d Occupational Mpdirino Department of Community Medicine M ount Sinai School of Medicino New York, New York PETER DECKERS Departments of Pathology and Surgery Hartford Hospital Hartford, Connecticut ABSTRACT, The etiology of human breast cancer Is unknown; accepted risk factors, e.g., menstrual, re p ro d u c e , and family historic*, ore implicated in less than half of II Various hafagenated hydrocarbons--Acting as either co-cardnogens or promoting agents --which are derived from rii* environmenr and are amccniratcd In human fatty Mlcr&i, may also ptay a role In breast cancer risk. A pilot study was undertaken to measure and comparo levati of chemical residues in mammary adipose tissue from women wRH malig nant and nonmalignant breast d in a . Elevated levels of polychlorinated biphiiyfa, bis (4-chforophenyQ-l,l diehlomethene, and h{sC4-diIoropheny0-1,1r1 trichloroethane were found In fat samples from women with cancer, compared with those who had ben ten breast disease* These results, although preliminary, suggest a role for environmentally derived suspect carcinogens fa the genesis of mammary carcinoma WIDESPREAD ENVIRONMENTAL CONTAMINA TION has occurred with several pesticide residues of the halegetuted hydrocarbon family, Including dlchlorodiphenytolchloroethane (DDT), dichlorodlphenyldi* chlcro-othylcne (DDE], polychlorinated biphenyls tPCBs], polybrominated biphenyls (PBBs), benzene lexachtorid? (BHQ, hexachlorobenzene (HCB), diet- drin, and dilofdane. These highly persistent, lipophilic, environment! chemicals have been detected world wide In wildlife and In humans.1These c h e rn M s cause cancer in animals, but evidence for human carcino genicity Is scant. It has also been reported that these compounds possess estrogenic activity,2 suppress Im mune function,* and induce hepatic microsomal en zymes.* These observations, and early studies of breast tumor tissue,5 suggest that these com pounds may in crease the risk for some cancers, Including mammary carcinoma. Another study found no difference In DDE VUtrh/April 1992 [VqI. 47 (No- 2)] 143 M A R - 2 5 - 9 2 LJFT1 5 . ? . ^ - , , . S J f j - h h i - *z P J? 0 3 .'2<i,-62 19:23 3 0 . J8 9888 ASA/DOXCOM . . s e n b a m g .. ik R. 03 0003 and PCBs bitw een cases in - 14) and controls tn - 21), but the numbers were not large, ana the findings deserve corroboration * ,. We undertook a pilot study to determine whether levels of several chemical residues, including DDT and PC8s, were higher In mammary adipose tissue from women who had malignant breast disease, compared with mammary adipose tissue from women who had benign breast disease. Materials and methods Patient population. The study population was de rived from a series of 50 Caucasian females seen for in itial surgical evaluation of & palpable breast mass or mamrnographTc abnormality a t Hartford Hospital (Hartford, Connecticut) from May through September 1987 After frozen section consultation was completed, 0.5 g of grossly nondiagnostic fat was separated from the biopsy or mastectomy specimen, coded, and stored at - 7 0 C in prewashed (detergent, water, acetone, hexane) vials (American Scientific Products) until for warded for chemical analysis. The samples were col lected consecutively by one pathologist in a nonbiased fashion. The only selection criterion was the presence of gross fat In an amount considered adequate for chemical analysis. Informed consent for tissue ex amination by the laboratory was obtained in each case. Routfoe Informed consent was adequate for the pur pose of this study because the chemical analyses were performed on fat destined to be discarded without fur ther processing. Twenty-three fat samples were obtained from wom en who had mammary carcinoma, and the remaining V samples were from wom en w ho had only benign disease. Fat samples from 40 patients In this Initial group (20 benign controls and 20 malignant cases) were chosen for chemical analysis; therefore, the ages for the two groups were similar. Patient height, w eight and smoking histories were obtained from the medical records or from brief telephone Interviews. Other data, such as specific dietary histories, were unavailable. Hlstopathofogy. Histologic sections of all breast le sions were prepared from formalin-fixed, paraffinembedded tissue, stained with hematoxylin-eosln, and examined by one of the authors CAR). Benign disease was classified according to the 1985 Pathologists' Con sensus Statement,' Carcinomas w ere classified as In situ and/or invasive ductal or lobular, and all patients were staged according to the cancer criteria of die American Joint Committee. Chemical analysis. Standard techniques were used to analyze 0.5-g coded samples for hexachforobenzene; nexachforocydohexane (lindane); three chlardano residues (heprachloroepoxkfe, oxychlordane. transmonachloi},* DDT residues (bbl4-chforOpfnyil-1,i dienforoethene fp,p#-DDC), bls[4-chlorophenyiJ-T,'I,T trichloroethane fop'-DDT]); and PCBs. The method was modified from that reported9; 1:1 dlchloromethane-cycloh exape was used for autom ated gel permea tion chromatographic lipid removal, arid ? Flarisil col umn was used to enrich PCBs and pesticides in two 144 fractions. PCBs were calculated as Arcelor 1260 (peaks with retention time greater than that for p.p'-D D by the method of W ebb and McCall,1 C o n cilia tio n s were reported on the basis of both lipid weight (uncorrected) and wet weight, corrected for recovery of an in ternal standard (a(cirin), because variance introduced by the lipid weighing step and by the clean-up steps was significantly reduced by this calculation (F test for among-/wltbin-run variance) for all chemical residues, except HCB. Results for 14 quality control samples (he-, chicken fat fortified with pesticides and Arodor 1260-PCBs) were, based on lipid weight, 74 22% for HCB and 99-109 5-19% for other residues; and on the basis of wet weight, corrected for aldrin recovery, 59 18% for HCB and 60-88 12-20% for other resi dues. Lipid constituted 74 5% of the wet weight for the 40 human samples. In addition to the t tests, comparisons w ere m ace for which nonparametric techniques (Wilcoxon rank-sum) were used, but the results were very similar. The me* dians and geometric means w ere similar to th e arith metic means. Calculations and statistical analyses were performed with the Statistical Analysis System (SAS In stitute, Cary, N Q at the City University of New York Computer Center. Results Patient population. The mean egos w ere similar for the two groups (63 y for cases, 59 y for controls), but age distributions were different; tne standard devia tions, medians, and ranges for the cases and controls were 13, $4, 36-B6 and 8, 58, and 45-76, respectively. More female controls had a history of current or past smoking (15 of 20) than did cases (6 of 20). Average heights and weights, including Q uetelet's index (w/h5), were almost Identical for th e two groups, i.e., average height and weight for cases (n " 18) w ere 161 5 cm and 68 14 kg, respectively, and 162 6 cm and 70 15 kg, respectively, for controls (n - 78). (Data were not available for four persons.) Hbtopathofogy. The majority of controls showed nonproliferative fibrocystic changes, which w ere some time macrocysts or combined with small fibroadeno* mala. Other histological diagnoses Included physio logic changes, proliferative fibrocystic changes without atypla (Including one solitary papilloma), and solitary fe bmadenoma. Tliere were no examples of atypical hy perplasia in this series. Sixteen of 20 (60%) cases had ductal carcinoma with an Invasive com ponent, 2 of 10 (10%) had invasive lobular carcinoma, and 2 (10%) had only in situ (ductal) disease. Oim ropaihoiogtc stages were Tis-2/20, 1-7/20, lf-8/20, and 111-3/20. O ne fourth of the cases had a history of contralateral carcinoma. O ne patient who had Invasive ductal carcinoma also had lobular carcinoma in situ in the same breast and 3 of 20 patients had 2 discrete breast tumors. Chemical analysis. Mean concentrations o f PCBs and p,p'-DDE were 50-60% higher in tissues of women who had breast cancer (wet weight basis, Table I). Levels of p,p'~ODT were also elevated among cases. The differences w ere statistically significant, regardless of Archives of Environmental Health o ij M A R - 2 5 - 9 2 MED 1 0 = 2 7 C A LL BEFORE SENDING i . E\ P . 04 Tab! 1---F&trrffte and PCs Concenintiores in Mammary Adipose Tissue C^g/g) from Cases will ertii* Capeer (n m 20) and from Controls with Bqnfoi P isw e fn 20) vr.-wwjsysswcrr-=vw Posted! Of PCS concentrations Cet Rarifc Controls Jf* SD Range um HC5 s * OC TN DDE DDT K to Wet weight heft. COtr&ttd h r rsavsrfy fng/jp 23* a 16 i 50 e r * 37 1 B7? ft 1 203 irs * las 1 669 * 894 13- 4J 41- 203 24- 182 337-4982 60- 6B6 733-4 674 2 0 * 10 5 / 49 9 6 80 1 174 630 14 49 1 105 424 12- 54 25- 249 2?A- 39* 237-2 2*6 4 4 - 248 5SZ-2 6CI9 .32 .22 .$$+ .04+ .05+ 02r HCS HX + OC TN DOE bor PCfiJ 28 * 11 136 52 103 43 2 200 1 1 470 216 174 1 963 927 Upfd basis, unM?cied fcg/& 76- 61 66- 243 38- 197 425-6 396 72- aai 627-4 562 26 11 121 53 116 87 1487 642 140 73 T395 t 469 14- 80 33- 278 S3- 439 303-3 674 4 2 - 405 823-2 87# .54 .36 . .49+ " j0 7 + .12+ >02t Nates HCH -i ftexachlor&tenzene, HX + OC - hepiacWoropcxIde and oxychlonfane the sum), Tn wanviiun*. chtor, ap'-D D E * fe1fi(4<+t6nCiit:nylMi 1-cJlchtoroethene, p.o1-DOT i- W8l4-chloroj)hny()il1(l `tftcWo'WrtK<.`i&. end m polychlorinated biphenyls. 'TwMalted probability for t, iro* r9unequal variant. whether parametric or nonparametrlc (Wllcoxon ranksum) methods we* used. O ther pesticides were not significantly different in eases and controls. There were differences In age distribution between cares and con trol#; therefore, logistic regression model# were used to confirm the relationship between level# of PCBs, DDE, and DDT and breast cancer status with respect to po tential confounding factors (age, Quetetet's index, smoking status), Even though smoking is not accepted as a significant risk factor for breast cancer, It was in cluded because of the marked difference between cases and controls in our study. In these models, PCBs remained significant, DDE remained significant when smoking was not in the model (with smoking, p -* .06&), end DDT was no longer related to cancer status. Because smoking I# not considered a major risk factor for breast cancer, w e conclude that the logistic regres sion models support the simple statistical tests that show differences between cancer and poncancer tissue levels of DDE abd PCBs. Logistic regression coefficients are shown in Table 2. The coefficients for DDE end PCBs ire approximately 0.001, which suggests that a 10-ppb increase fn tissue level corresponds to a 1% In crease In risk of breast cancer. However, because other important risk factors for breast cancer were not studied, there are uncertainties in interpretation of these findings, which require verification Jn a larger study designed to control for other known risk factors. Discussion Breast cancer is th? most prevalent cancer among American wom en.11 However, the Identified risk fac Miifdi/AprU1992JVol, 47 (No. 2)] tors account for considerably less than half of breast cancer In the United States. Breast cancer risk has been correlated with reproductive factors, e.g., early age at menarche and late onset of menopause, which sug gests that hormonal factors may be Involved. Genetic markers and dietary contributions have been impli cated in some studies. Variations In rates of breast cancer internationally are not readily explained by these factor?/* and, therefore, other environmental facto/?, such a# exposure to carcinogenic chemicals, may be part of the explanation. Differences in such ex posures might account for disparate rates of breast cancer, e,g., between racial groups and between coun tries with similar dietary profiles. The difference in our findings from those of Unger ct a i / may result from chance or from differences In the study groups, e.g., nationality or other confounding factors. The finding of higher tissue levels among cancer eases may also . signify a redistribution of chemicals to the breast during the disease process. These caveats, and the preliminary (or pilot) nature of our findings, necessitate further Investigation of the question of environmental chemicals and breast cancer with better age matching and attention to other factors known to contribute to breast cancer, such a# parity, age, and menopausal status. Because these chemicals are probably promoters or induce, rather than direct ] 7 carcinogens, their Involvement, If any, in human 1 * cancer may be part of a complex array of genotoxle 1 and epigenetic effects. In light of Increasing rates of breast cancer1* and our llmlteo knowledge of the causes. Investigation of car cinogenic environmental chemical exposures Is a 145 M rtR --2 5 -- * * W ED 1 0 = 2 v 61^. CPLL B EFO R E SEHDXMG 4-k T a h iti.--LugUdc Regression CotffieltpU tor Breast Cancer In C u ts and Control Residue rniecept Age Age 2 Residue Smoking DDE 29,052' --1,0SQ" 0.00896* q^xwast - 2^36^ -1.162' 0.00927" 0-00122* -2.662* PCB 3.552* -1.059" O.OCS64' 00022* - 3US1" -1.088* 0,00859* 000234* --1.658t .vosrT AiQtf V?>3^5; residue UIn parts per btlDnn: and smoking ever 1, ra w m 0. Queteteds lndc>h y xtX v y ^ t $4f>ftlcdt significance In these models. The standardized coefficients were ppf^nstsiy 03. 'ft < JOS. *<-<37*, F1- 0 5 pithTiisiiTg sv-stae to explore in our efforts to further e^xiriLve cte origins of this disease. Supported In pan by National InnTturo of Environmental Health Sciences grant ESC0928, The authors thank Dr. Robert Rippey for oatlsttcU conjugation end Ms. Marilyn Rivera for technic*! assistance. Submitted for publication November 2d. 1390; revised; accepted for publication June 26, 1991. deques* for reprints should be sent to: Dr. Frank Falck, Jr., Department of Ophthalmology,, University of Michigan, 1000 Wll Street, Ann Artwr, Ml 401OS. V ***I**P References 1. Murphy R, Harvey C. Residues and metabolites of Selected per sistent halogcrtated hydrocarbons In blond specimens horn a general population survey. Environ Health Hetvpecl 60:115-30. 2. Meson PR, Schulte CJ. Interaction of o,p'-DDT with the estrogen-bindln* protein (EBP) of DMBA*Inducod rat mammary tumor. Res Comma Chem Pathol Pharmacol 1981: 33:119-26, 3. Exon JH. Kerkvlle? Nl, Tgleon PA, ImmunoLovicity nf car cinogenic pesticides and related chemical*, J Environ 5 a H??Wl Part C Environ Circin Rev IS87; C5:73-120l a. Partwcon a, %ertson U Safe L Safe S. Pofychlorlrwed bt^tws^b as hidveer? of hepatic midrowmal er<zymes*. structure. foJoSL Cbem gid idee 19BO: 3%27*-d5. 5. iVasjafflar M, Ncgueiia DP, Tomails L Mar* A?, Shibata H, Arie G, C ue Sy Waxserman D. Oiganadtlcttne compounds In neoptasde end adlacenr apparently nomuJ brew: tune. Bull EnAon Conam Tdcoi 1976; 15:470-64, 6- Unger M, JOeor H, BUchert-Toft M. Olsen J- Clausen J, Organochlofrte compounds In human breafl & from deceasedwithout breast cancer and In a biopsy materiel from newly diagnosed patients undergoing breast surgery. Environ Res 1934s 3434-28, 7. Mutter RVP. Is fibrocystic disease of the b re w precamrerousl Arch Pathol bib Med t98fi; U 9:171. a. Beahrt OH, Myers MH, Ed. Manual for staging a n /-, 2nd ed. America Joint Committee on Cantor. Philadelphia. Pa: 10 Llppltv COtV 1983; 127-34, 9. Wolff MS, Fischbelft A, Thomtbn }. Rice C URs R. Sel*koff IJ. Body burden of polychlorinated biphenyls among persons employed In capacitor manufacturing. Ini Ardh Occup Environ Health 1982) 4*199-2. 10. Webb RG, McCall AC, Quantitative PCB standards for electron capture MS dittmetography. J Chromat Sci 1973; ll;366-73. If, Kelt/ Jb Bericowitt GS. Breast cancer epidemiology. Con ftej 1983; 40:5615-23. 12. Willett W, The Search for the causes of breaA and colon eencer, Nature 1989; 338:309-94. 13. Class AC, Hoover RN. Rising Incidence of breast cancer; relattofehlp to stole and receptor sam s, J Natl Cancer im 1990; 82: 93-96, 146 Archives of Envfaontnenftf Health