and ‘8FDGin nonHodgicin's lymphoma (14). The purposeof this research was to examine the relationship between“C-methionineuptakeand the flow-cytometric parametersin NSCLC.

Methionine is either utilized for protein synthesis orserves as the biological methyl donor for the methylation ofDNA, transfer-RNAand other compounds (transmethylation) after formationof 5-adenosylmethionine (SAM). Therelative magnitude of these alternates is not well understood. In vitro experiments have shown that cultured celllines of human tumor (15,16) and activated lymphocytes(lymphoblasts) (17), are extremely dependent on methionine supply in the culture medium because of the reducedactivity of methionine synthesis from homocysteine, contrary to all other normal cells. Bustany et a!. (18) reportedthat@ ‘Cmethionineaccumulation in brain tumors was related to its grade. Leskinen-Kallio et al. (13) reported thatthe uptake of “C-methioninein breast cancer was associated with a large S-phase fraction measured with flowcytometry.Leskinen-Kallio et al. (14) found the correlation of“C-methionineuptake to S-phase fraction was higherthan that of ‘8FDGuptake to the S-phase fraction innonHodgkin's lymphoma. Fujiwara et al. (19) reportedthat the ‘‘C-methionineuptake in human lung cancers varied among its different histological types.From the above reports it was expected that ‘‘C-methionine could be used for visualizing lung cancer, withthe intensity of uptake rate proportional to the tumorgrowth rate.

The abnormalDNA content (aneuploidy) and proliferative characteristics as shown by S-phase and G2IM-phasecell population can be determined from the tumor cellsuspension. It is usually prepared from freshly frozen tissue specimens. The paraffin-embedded material can be employed also, but it is only good for the ploidy analysis(20,21). It has been advised that the cell-phaseanalysiswith the latter material be avoided. In this work, flowcytometiy was performedusing paraffinblocks and freshlyfrozen specimens for ploidy analysis. Only freshly frozen

Carbon-i1-methioninePETscanswereobtainedfrom 24 patientswith non-small-celllungcarcinomafor whom surgicaltreatmentwas considered.The tumor masswas VISUalIZedwith deardelineation. After PET scanning, the tumor was removed bylobectomy or pulmonectomy. The tumor tissue was first processed to @dtumor cell suspensions and then subjected toDNAflowcytometry.Compatisonbetween11Cuptakerateandflow-cytometricdata gave the following results: 11Cuptake rateinthe tumorcorrelatedwellwiththecellularDNA content(DNAindex)of tumorcellsat the restingstateof celldivision(GO+Gi-phase) (r = 0.67).The correlationbetween 11Cuptake rateand S-phasecell percentagewas markedtyhigh (r = 0.76), andthecorrelationbetwee11CuptakerateandS + G2/M-phasecellpercentagewasextremelyhigh(r = 0.86).ftwascondudedthatthetumoruptakerate@ 1C-methioninewasrepresentiveoftumorgrowthratein thistumortype.

J NuciMed1993;34:1886-1891

he clinical staging of non-small-cell lung carcinoma(NSCLC) depends on the TNM classification. Only theanatomical (morphological) characteristics of the tumorsare considered, but the biologic parametersof tumortissueare not included in this staging procedure. Ploidy and proliferatingactivity of tumor cells analyzed by flow cytometry are widely accepted as good indicators of tumor malignancy (1—9).The positron-emitting radiopharmaceuticals,L-[methyl-―C]methionine, “C-methionineand ‘8F-2-fluoro-2-deoxy-D-glucose (‘8FDO)are widely used for tumor imaging,and the uptake rateof these tracers in tumorshave been studied in comparison with flow-cytometric parameters: with 18FDG in head and neck cancer (10—12),“C-methioninein breast cancer (13) and “C-methionine

ReceivedJul 28,1992;revisionacceptedJun.24,1993.Forcorrespondenceor reprintscon@ ToshihkoHera,InternationalMedical

CenterHospital,1-21-1Toyaria,Shir@ukuku,Tokyo,J@an.

1886 TheJournalofNudearMedicine•Vol.34•No.11•November1993

PET Imagingof Non-Small-CellLungCarcinoma with Carbon-11-Methionine:Relationship Between Radioactivity Uptake andFlow—CytometricParametersHideki Miyazawa, Takashi Arai, Masaaki ho and Toshihiko Hara

Departments ofSuigery and Radiolo@j@,National Nakano Chest Hospita4 Tokyo, Japan

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Patientno.*Age Sex Diagnosist pTNM*

—

*Corr@1_ndstoTable2.tH@gIc@Jdiagnosisdeterminedon surgicallyresectedmaterial:

Meno = adenocarcinoma;Squamous= squamous-cellcardnomaandMenosquamous = adenosquamouscardnoma.

@Pathdo@ThM classlflcatlon.•Pathdo@stage.

specimens were used for the S-phase and G2IM-phase cellfraction measurements.

PA11ENTSANDMEThODSPatients

PatientswithuntreatedNSCLCunderwentthisstudy.All 24patients were treated by tumor resection within 2 wk after thePET study. Twentyof the patientswere male and fourwerefemale. The mean age was 62.3 yr (range, 46—82yr). All patientshada tumor>2 cmdiameteron CFfilm.Morphologicclassificationof thebronchogeniccarcinomaswasdoneaccordingtoWHOcriteria(1981)(22).The tumorswere comprisedof 12squamouscell carcinomas, 10 adenocarcinomas, and 2 adenosquamous carcinomas.Theageandsex of thepatients,aswellas thehistological characteristics of the tumor, pathological TNM classificationand staging of each patient are summarized in Table 1. Stagingwas determinedby the guidelinesof the InternationalStagingSystem for LungCancer(UICC 1987)(23). Informedconsent wasobtainedfrompatientspriorto eachPETstudy.

ImagIng wfth Carbo@11-MsthIonk@A whole-bodyPETscanner,HeadtomeIV(Shimadzu,Kyoto)

(24), was used to image ‘1C-methionineuptake. For emissionscanning, we customarily employ a 6-mm spatial resolution in apatient cross-section by adjusting the Butterworth filter level.(During transmission, the spatial resolution was adjusted to 12mm.) The field of view was 512 mm in diameter. The slice thick

TABLE 1PatientCharacterisbcs

ness of direct and cross planeswas 11mm. The make-upof ourmachine supplies five slices simultaneously with a slice interval of13mm(witha totalof 63mm).Duringimagereconstruction,theimage matrix was constructed from data in the central part of thefieldof view (256x 256pixelsof 1mmsquareon therealscale).The total numberof counts for the transmission scan was morethan6 X 106,andat least 1.5 x 1O@fortheemissionscan, forasinglefieldof view.

Emissiondatawereacquiredusinga respiration-synchronizedgating device we fabricated which is connectable to the inputtriggersystemofthe PETsystem.It minimizedthe partialvolumeeffect caused by respiratory movements; it was monitored by abody impedance pletysmograph (“Respi-trace―,AmbulatoryMonitoring,NY)whilesensingchangesin the patient's abdominalcircumference.Radioactivitydatawereaccumulatedonlyduringtheexpiratorystateforthefirstthirdof therespiratorycycle.Thetransmissionscanwas performedwithoutrespirationgatingwhilethepatientwas askedto breatheshallowlyduringscanning.Correct positioning ofthe patient, placed in the supine position on thePETcouch,was achievedwithreferenceto chestx-rayandCTfilms, and by observation of the transmiss@n image.

Carbon-11-L-methioninewas synthesizedby Comar'smethod(25) with a slight modification using L-homocysteine thiolactoneinstead of L-homocysteine and “C-methyliodideduring transmission scanning. Carbon-11-methionine(about 10 mCi (370 MBq))was theninjectedby bolusintothe antecubitalvein andflushedwith saline. Data were collected at 0—2mis and 10—20mm, sep.arately, with respiration gating and were decay-corrected to timezero automatically. The first time-frame image showed the bloodpool imagethat facilitatedrecognitionof the anatomicallocationof bloodvessels in themediastinum.Thesecondtime-frameimage(laterimage)showedtheareaof 11C-methionineaccumulationin the tissue. The “Cactivity trapped in tumors during the first 10mm remained almost constant for 1 hr with decay correction.

The appropriatetimeperiodfor tumorimagingas describedabove was determined in a preliminary experiment by measuringthe time course of radioactivity of lung tumors (an adenocarcinomalocatedin thepulmonaryapexanda squamouscell carcinomalocatedin the hilusof the lung),withoutemployingtherespiratorygating.We selected these tumors expectingthat theinfluence of respiratory motion to the location of the tumor mightbe minimal. The radioactivity in the tumor remained unchangedbetween 10mm and 60 min after bolus injection of “C-methionine(Fig.1).

Thecumulativetumorradioactivitywas calculatedin the region of interest (ROl) placed over the later image. The “Cuptakewasquantitatedasfollows.First,we displayedthelaterimageona CRT screen to see the contour of tumor; the color displayenabledtheradioactivityconcentrationto appearindifferentcolors. Next, we placed a ROl, corresponding to 1.2 cm2 in bodysize, over the most radioactive area—usually the center—andobtainedtheradioactivityconcentrationintermsof cps/cm@.Carbon-il uptake was expressed by the differential uptake ratio(DUR) (19) as follows:

DUR - Tissue radioactivity concentration

- Total injected dose/Body weightS

The total injecteddose was determinedfromthe well-countercount of the injected solution after appropriate dilution, correctedby the cross-calibration factor between the well counter and PET.

@*156FAdenoTIN2MOlilA277MSquamousT1NOMOI359MSquamousT3NOMOlilA469FMenoT2N2MOlilA566FAdenoT1NOMOI661MSquamousT3N2M1IV782MSquamousT4N2MOhUB874MSquamousT2NIMOII975MSquamousTINOMOI1047MAdenosquamousT4N2MO1I1B1160MAdenoT3NOMOlilA1250MAdenosquamousT3N1

MOlILA1360MAdenoT1NOMOI1457MSquamousTINOMOI1546MAdenoT2N1MOII1667MSquamousT2N2MOlilA1755MSquamousT3N1MOlilA1867MAdenoT2N2MOlilA1973MSquamousT2N2MOhILA2071MAdenoT2NOMOU2155MSquamousT3NOMOlILA2260MAdenoT3NOMOlILA2360MSquamousT3NOMOlILA2446MAdenoT2N1MOII

Non-Small-CellLungCaranomaPETw@ 11C-Methionine•Muyazawaat al. I 887

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Table 2 is the list of “C-methionineuptake (“C-DUR),DNA ploidy, DNA index and the percentageof cells in theS-phase and G2IM-phaseofthe cell cycle. The “C-DURintumor was always high and demarcated sharply from thesurrounding tissue, as seen by direct observation of theresected lung. Figure 2 shows a typical PET image.

There was a significantdifference of “C-DURbetweenStage I (mean ±s.d., 3.88 ±0.47) and Stage II (5.28 ±0.12, p = 0.001) and between Stage I and Stage lIlA (4.91±1.14, p = 0.025). The 11C-DUR for Stage IIIB (n = 2)was 3.75 and 3.98, and 4.43 for Stage IV (n = 1) (Fig. 3).The cell type differenceof NSCLC, however, did not bringabout any significant difference in the “C-DURvalue.

Of 24 cases examined, DNA diploidy was found in fourwhile the rest showed aneuploidy with the DNA indexranging from 1.29 to 2.36. There was no multiploidy. Carbon-il-DUR ranged from 3.33 to 3.85 (3.62 ±0.22) indiploidy cases, and 3.46 to 8.05 (4.78 ±1.00) in aneuploidycases. The difference between the two groups was signiflcant (p = 0.0072). The correlation between “C-uptakerate, “C-DURand cellular DNA content at the restingstate of cell division was also significant (r = 0.67, p =0.0003, n = 24) (Fig. 4).

The composition of cells in different stages of the cellcycle was measured in 10 cases, using the freshly frozenspecimens. The relationship between the cell-cycle phasesand 11C-DUR is as follows: The percentage ofS-phase cellscorrelated with 11C-DUR(r = 0.76, p = 0.010, n = 10)(Fig.5). A marked correlation was noted betweenthe percentage of proliferativecells (S + G2/M-phase cells) and “C-DUR (r = 0.86, p = 0.0013, n = 10) (Fig. 6). On thecontrary, as a logical consequence, correlation betweenthe sum of percentage of resting cells (GO)plus proteinbut-not-DNA-synthesizing cells (Gi) with “C-DURwasinverse (r = —0.86).

Although the purpose of this study was to analyze therelationshipof “Cuptakerate in tumorsto the proliferativeactivity of the tumors, we simultaneously examined theeffectiveness of the “C-methionine method as a means ofdetecting tumor metastasis to the hilar lymph nodes. Carbon-li-methionine PET gave positive images (with DUR>3) in 12 lymph nodes (three patients), which appearednormal in size on CF film but were proven to be a tumormetastasis pathologically (PET true-positive). On the otherhand, “C-methioninePET was false-positive in only twolymph nodes (one patient) of 109CT-enlarged lymph nodes(27 patients). Following is the description of the only patient with false-positive lymph nodes by PET. This patient(Table 1, Patient 2) had moderately differentiated squamous-cell carcinoma in the right lower lobe. The CT filmshowed enlargement of pretracheal and subcarinal lymphnodes. With “C-methionine,the DUR for the tumor was3.33, and the DUR for the lymph nodes was 3.59 and 3.92,respectively (Fig. 7). Sternum and vertebra bone marrowalso exhibited an extraordinarily high uptake in this case.

5

4

3

0

U 10 20 30 40 50 60Minut.s

FiGURE 1. Typ@ bmecourseof11C-methkxiineuptakeinlungtumors.Carbon-i1-OURis thetumoruptakeof 11Cexpressedbytheratioofthetumorconcentrationof11Ctothehypotheticaltissueconcentrationof 11Cwhere the injected11Cis supposedto bedistnbutedunifomilythroughoutthe whoie body.S = adenocardnoma:0 = squamous-eelcarcinoma.

DNA Flow CytometryThe tumor samples for flow cytometiy were taken from the

resected lung in order to pick out the tumor tissue from the sameposition as used for the DUR measurement. Fourteen of the 24samples were taken from the paraffin-embedded blocks and 10weretakenfromthefreshlyfrozenspecimens.Thesampleswereexamined histologically and determinedto be free from contaminationwithnormalpulmonaryparenchymalcells.

The following procedure was conducted in collaborationwithOtsuka Assay Laboratory, Tokuyama City. When paraffin-embedded blocks were used, four or five 40-sm thick slices weretreated by the method of Schutte et al. (21). Normal pulmonaryparenchymal cells were used as an internal standard. For thefreshly frozen specimen, a 200-mgpiece of tumortissue stored ina deep freezer was cut into smallpieces and placed in phosphatebuffered saline. Human normal monocytes were used as a standard. The sampleswere stainedwith propidiumiodide.

The nuclearDNA contentand its histogramwere analyzedwith a FACSCAN/Cellflt DNA system (Becton-Dickinson, CA).The tumor cell preparationexhibiting only a single GO/Gi-peakwas regardedas diploid.TheDNAindexwas definedas theratioof aneuploid GO/Gi-peakchannel number-to-diploidGO/Gi-peakchannel number. Histogram samples with a variation coefficient

<8%wereconsideredgoodqualityandweresubjectedtoanalysis. Thecountof S-phaseandG2IM-phasecellswerecalculatedby the sum of the broadened rectangles (SOBR) method (26) oftheCeilfitDNA system.

All the specimens were used for ploidy analysis. Only thefreshlyfrozenspecimenswere usedforanalysisof S-phaseandG2/M-phase cell population, because the paraffin-embeddedblocks are not suitable (21).

Statistical AnalysisMeanandstandarddeviationwerecalculatedforeachgroup.

The differenceof group means was examined with a t-test. Scattered plot was analyzed by linearregression analysis. The coefficient of correlationwas analyzed by t-test.

1888 TheJournalofNudearMedicine•Vol.34•No.11•November1993

RESULTS

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Ao@pemmatarPatient

S-phase S +GVM-phaseno.11CDUR Ploldy DI cells(%) eels(%)Speclmen@1

3.60 D 0.98 ——p23.33 D 1.00 ——p33.71 D 1.00 1617f43.85 D 1.00 1420f53.46 A 1.29 ——p64.43 A 1.33 1727f73.98 A 1.36 ——p85.35 A 1.42 3155f94.23 A 1.46 ——p103.75 A 1.50 ——p114.71 A 1.52 3136f124.10 A 1.63 ——p133.46 A 1.66 ——p144.46 A 1.70 ——p155.38 A 1.78 ——p168.05 A 1.79 8791f175.16 A 1.80 ——p185.29 A 1.85 1625f195.31 A 1.86 ——p204.31 A 1.90 1221f214.65 A 1.91 2227f225.38 A 1.93 ——p235.05 A 1.97 ——p245.12 A 2.36 2835f*Specimars

usedforflowcytometryp = paraffin-embedded(n= 14)andf = freshlyfrozen(n=10).licDUR = 11Cdifferentialu@Mkeratio,Dl = DNAindex,D = d@loId,andA = aneuploid.

8.0•Mw@±$.@7.00

I

.9—6.0

50•

4.0

3.0t•*

Sfl±OI2

)if3@±047

VA@14,I±,,4

#1

@,1 :i3S7±0122.01.0I

III@-I I

(n6) (n=3)IAlB

(n@12) (n2)N(n1)Pathological

stage

TABLE 2Carbon-i 1 Uptake After lnjec@onof Carbon-i 1-Methionine, DNA Pkkty, DNA index and Cell Cyde DiStribUtiOn

The resected lymph nodes were examined histologically.They were somehow differentin appearancefromthe commonly seen lymph node hyperplasia induced by tumorreaction in that they were occupied by copious lymphoidfollicles containing large germinal centers with prominentmitotic activity. Histiocyte clusters were found sporadically between the fofficles. Tumor metastasis was not cvident in the lymph nodes.

DISCUSSION

In this study, DNA ploidy and the percentage of proliferative cells of NSCLC were compared with “C-methionine uptake rate. We used only freshly frozen specimens(10 samples for the cell-cycle analysis), because paraffinembedded blocks would contaminate background debris

and cell aggregates (21). The mean percentage of S-phasecells in our specimens (27.4%) was higher than those ofother reports (27—30).The highest percentage of S-phasecells (87%)observed in Patient 16 corresponded with thehighest accumulationof “C.

In order to keep anatomical correspondence betweenflow cytometiy and PET, we worked carefully in cuttingout the tumor tissue from the most radioactive area in the“C-methioninePET image and rendered it to flow cytomdry. This was facilitated by the respiration-synchronizedscanning method, which the apparently stopped respirationreduced the partial volume effect on the tumor image.

FiGURE2. (Left)PETvasctiarimage0-2 mmafterinjection. --Svc=supenorvenacavaandAo=ao.ticarch.(R@ht)PETimageFiGURE3. Comparisonof11C-DURwfthpathdoglcalstages.of lungtumor (lateimage)10—20mmafter Injection. E@hvaluerepresentsmeanand standarddeviation.

Non-Small-CallLung Carcinoma PET w@i11C-Methionine•M@razawaSt al. 1889

,

@ &

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90 9.0

1.0

7.0

fr34.0 •0

3.0

r . 0.662.0 p.0.0013

1.0

0.1 , , I I I@ I@@@ 40 SO 66 70 66 90 100@'5+01 /M-phau cilia

0@

•o@.0@a

f •0.67

p •0.0003

to•

7.0

60

@ to

@ 40

3.0

1.0

0 †1̃.0 2.0DNA Indsx

FIGURE4. Conelatlonbetween11C-DURandDNAindexfor24NSCLC.O = Stagel;®= StagelI;c@=Stagelhl;and• = Stagelv.

When we used only freshly frozen specimens, a veryclose correlation between the percentage of proliferativecells and the “C-methionineaccumulationwas observed.If we included (not shown in Results) both freshly frozenspecimens and paraffin-embeddedspecimens (n = 12 instead of 10), the correlation coefficient dropped from 0.76(p = 0.010) to 0.54 (p = 0.070) for S-phase cells, and from0.86 (p = 0.0013) to 0.75 (p = 0.0052) for S + G2IM-phasecells.

It is well known that the biosynthesis of methionine isdeficient or extremely reduced in malignant tissues (15,16).It follows that the malignantcells have high demands forexternally added methionine. Once the “C-methioninemolecule is taken up in the cell, it will undergo one of thefollowing alternative pathways: integration into protein orincorporation of its methyl group into DNA (addition ofmethyl group to purine and pyrimidine bases of alreadyformulatedDNA molecules) and other compounds.

Proteins within cells are in a continuous steady state ofsynthesis and degradation(and excretion). Ishiwata et al.(31) reported that, in rats bearing Walker 256 carcinosarcoma, the major tumor cell component for “Cincorporation after injection of “C-methioninewas proteins.

3.0

FiGURE6. CorrelatIonbetween11C-DIJRandproportionofS+G2i1@A-phasecells(n= 10).0 = StageI; @D= StageII;G = StageIII;and•= StageIV.

Our study in humans demonstrated a close correlationbetween accumulation of “qmethyl)-methionineand incremented cellular DNA content (aneuploidy) in NSCLC.There was also a close correlation between high methionine uptake in tumor and high percentage of proliferativecells in tumor, where the nuclear DNA is duplicating (Sphase), or the DNA content is already doubled (G2/M-phase). This observation seems to suggest a large contribution of transmethylation process to the “Cuptake inhuman lung tumors.

It has been verified hat nuclear DNA content and thepercentage of proliferative cell fraction are correspondingto the aggressiveness of human lung cancer and patientprognosis (2—6).Our study indicated that the measurementof “C-methionineuptakewould help evaluate the proliferation rate of lung cancer.

The lymphoblasts, either inflammatoryor immunoreactive in origin, are methionine-dependent (15,16) and areexpected to have a tendency for high methionine uptake.We observed this in one case of mediastinal lymph nodeenlargement caused by tumor reaction. This lymph nodewas characterized by a markedly high lymphoblastic proliferation.

Leskinen-Kallio et al. (14) reported that there was nocorrelation between “C-methionineuptake and S-phase

2.0

1.0

0@@@@ 66 70 60

S-phas. cilia

head

_@)@body

FIGURE7. Asingiecaseofpositiveuptakeof11C-methkxiineexhibitedby lymphnodesof nonspecificreadffvehyperplasia(histologicaltyproven).I = vertebra;2 = sternum;3 = myocardum;4= lung tumor (DUR, 3.33); 5 = pretracheal lymph node (DUR, 3.59);

6 = subcarinallymphnode(DUR,3.92).

FiGURE5. Corretationbetween11C-DURandproportionofS-phasecells(n = 10).0 = StageI;@D= Stagell;@ = Stagelii;and S = StageIV.

1890 TheJournalofNudearMedicine•Vol.34•No.11•November1993

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cell percentage in nonHodgicin's lymphoma. However, aproblem seems to be involved with the paraffin-embeddedspecimens they used for the flow-cytometric analysis, aswe previously mentioned. Inbreast cancer, they also foundan association of “C-methionineuptake with the size ofS-phase fraction (r = 0.77, p = 0.01) (13) using paraffinembedded specimens. They concluded that the accumulation of 1'C-methioninemay correlatewith the proliferationrate of breast cancer.

Kubota et al. (32) reported that the radiation responsemonitored by radiotracer uptake was similar betweenL-[methyl-'4C@methiornne and [6-3Hjthymidine, but verydifferent from the response with ‘8F-FDGand 67(j@in anexperiment with a rat tumor model combined with radiotherapy. This observation is suggestive ofa close link in themetabolism of methionine and thymidine in the rat tumor.

In summary, NSCLC was visualized with “C-methionine. In this tumor type, the intensity of “C-methionineuptake was strongly associated with the cellular DNA content and the extent of duplicating DNA (the latter representing the proliferative activity of the tumor). Carbon-limethionine imaging seems to be a reliable technique forattaining a noninvasive diagnosis of aggressiveness ofNSCLC.

ACKNOWLEDGMENTSThe authors would like to thank Mr. Toshiaki Abe of the

Institute of Physical and Chemical Research, Wako City, for hiscollaboration in fabricating the respiration-synchronizedgatingdevice; and Drs. Hitoshi Niino and Seiichi Serizawa for theirinterestand technical assistance. This work was supportedby theJapanese Ministry of Education, Science and Culture Grant-inAid for Cancer Research (02151072) and by the Japanese Scienceand Tecimologj@Agency.

REFERENCES1. Akin NB, Kay R. Pmgnosticsignificanceof modalDNA valuesandother

factorsin malignanttumorsbasedon 1465cases.BrJ Cancer197940:210-22k.

2. vo1n@M, Mattern J, Sonka J, et al. DNA distribution in non-small-cell lungcarcinomasanditsrelationtoclinicalbehavior.Cyto,nebyl985;6:348-356.

3. Vohn M, Drings P, Mattern J. Pmgnostic significance of DNA patterns andresistance.predictive tests in non-small.cell lung carcinoma. Cancer 1985;56:1396—1403.

4. ZimmermanPv, TawsonGA,BintNH, ParsonsPG.Ploidyasaprognosticdeterminant in surgically treated lung cancer. Lancet 1987@:530-533.

5. Tirinddlli-DanesiD, TeOdOnL, Mauro F, Ctal. Prognosticsignificanceofflowcytometiy in lungcancer: a five-yearstudy. Cancer 1987;60:844-851.

6. VolmM, HahnEW,MatternJ,etal.Five.yearfollow-upstudyofindepen.dent clinicaland flow cytometricprOgnOsticfactorsfor the survivalofpatientswith non-small-celllungcarcinoma.CancerRes 1988;4&2923-2928.

7. JoensuuH, KlemiPJ, KorkeilaE. Prognosticvalueof DNA plOidyandproliferative activity in Hodgkin's disease. Am I Clin Pathol 1988;90:670-673.

8. Ensley JF, Maciorowski Z, Hassan M, Ctal. Cellular DNA content parametersin untreatedandrecurrentsquamous.ceilcancersof the headandneck. Cytomet,y 1989;1O:334—338.

9. MerkelDE, McGuireWL PIOldy,proliferativeactivityandprognosis.DNAflowcytometiyof solidtumors.Cancer1990;65:1194-1205.

10.MumH, JoensuuH, AhonenA, KlemiP.Fluorodeoxyglucoseimaging:amethodtoassesstheproliferativeactivityofhumancancerinvivo.Corn.parisonwithDNAflowcytornetiyinheadandnecktumors.Cancer1988;61:1776—1781.

11. Minn H, Paul R, Ahonen A. Evaluation of treatment response to radiotherspy in head and neck cancer with fluorine-18.fluorodeoxyglucose.I NuciMed 198829:1521—1525.

12.HaberkornU, StraussLO, ReisserC,Ctal.Glucoseuptake,perfusionandcell proliferationin head and neck tumors: relationof positron emissiontomographytoflowcytornetry.JNuclMed1991;32:1548-1555.

13.Leskinen-KallioS. NagrenK, LehikoinenP.Ctal.Uptakeof “C-methionine in breast cancer Studiedby PET. An associationwith the size ofS-phasefraction.Bri Cancer1991;64:1121-1124.

14.Leskinen-KallioS,RoutsalaincnU, NagrenK, etal.Uptakeofcarbon-il.methionineandfluorodeoxyglucosein nonHodgkin'slymphoma:a PETstudy.INuci Med 1991;32:121l—1218.

15. HoffmanRM.Alteredmethioninemetabolismandtransmethylationincancer.AnticancerRes 19855:1-30.

16.JuddeJG,FrostP.Patternsofmethionineauxotrophyinnormalandneo@ticcells: the methionine independenceof lyInphOCytICmitogenesisand

low frequencyof the methionine.dependentphenotype in human tumors.CancerRes1988;48:6775—6779.

17. Kano Y, Sakamoto S, Kasahara T, et al. Methionine dependency of cellgrowthinnormalandmalignanthematopoieticcellsCancerRes1982;42:3090-309@

18. BustanyP, ChatelM, DerlonJM,CtaLBraintumorproteinsynthesisandhistologicalgrade:a studyby positronemissiontomography(PET)with“C-L-methionine.INewol Oncoll986;3:397-404.

19. FujiwaraT,MatsuzawaT,KubotaK, et al.Relationshipbetweenhistolo@ctype of primaiy lung cancer and carbon-11-L-methionineuptake withpositronemissiontomography.INuciMed 198930:33-37.

20.HeadleyDW,FriedlanderML, Taylor1W,Ctal.Methodforanalysisofcellular DNAcontentofparaffin.embedded pathologicalmaterial usingflowcytometly. JHLstOChen* C)'tochem 1983;43:3982-3997.

21. SchutteB, ReyndersMJ,BlijhanGH,Ctal. Flowcytometricdeterminationof DNA ploidylevel in nucleiisolatedfromparaffin-embeddedtissue.C)@tometiy1985;6:26—30.

22. World Health OrganiZatiOn.Histologicalaping of lw@ tumors, secondedition.Geneva:WHO; 1987.

23.HarmankP,SobinUi. TNMclassificithonofmalignanttuna@.Geneva:Union intumationale contre le Cancer, 1987.

24.lidaH, MiuraS,KannoI, Ctal.DesignandevaluationofHeadtomeIV, awhole-body @ronemissiontomograph.IEEE TransNuci Sd 1989;36:1006—1010.

25. CoinerD, Canon JC, MaziereM, MarazanoC. Labelingandmetabolismofmethionine-methyl.11C.EurlNuciMed 1976;1:11-14.

26. Dean PN. Data analysisin cell kineticsresearch. In: Gray JW, DarzynkiewiczZ, eds. TeCIUIIqUeSÜIceilcyckan4@sis.Clifton,NJ: HumanaPress;1987:207—253.

27.RaberMN,BarlogieB,FarquharD.DeterminationofploidyabnormalityandcellcycledistributioninhumanlungcancerusingDNAflowcytometly.PmcAmAsa:@c CancerRes198021:159.

28. Bunn P, SchlamM, GazdarA. Comparisonof cytology and DNA contentanalysisbyflowcytomet,yinspecimensfromlungcancerpatients.PmcA,nAssoc CancerRes 1980',21:160.

29. Olzewski W, Darzynkiewicz Z, Claps ML, MelamedMR. Flow cytometiyof lungcarcinoma:a comparisonof DNAstemlineandcell cycle distubu.tion with histology. Anal Quant C@ytd19824:%—94.

30. Teodori L, Thindalli.Danesi D, Mauro F, Ctal. Non-small.cell lung cardnoma:tumorcharacterizationonthebasisofflowcytometricallydeterminedcellularheterogeneity.Cytonietiy1983;4:174-i83.

31. IshiwataK, vaan,@ W, ElsingaPH,etal.Comparisonof L-[1.'1C]methionine and L-[methyl.―qmethionine for measuring in vivo protein syn.thesis rates with PET JNuclMed 1988;29:1419-1427.

32. KubotaK, IshiwataK, KubotaR, etal.Tracerfeasibilityformonitoringtumorradiot&rapy a quadrupletracer studywith fluorine.18-fluorodeoxyglucoseorfluorodecxyuridine,Umeth@.'@methionine,[6.3H@thymidine,andgallium-67.JNucIMed1991;32:2118—2123.

Non-Small-CallLung Carcinoma PET w@i11C-Methuonine•Miyazawa at al. 1891

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1993;34:1886-1891.J Nucl Med.   Hideki Miyazawa, Takashi Arai, Masaaki Iio and Toshihiko Hara  Relationship Between Radioactivity Uptake and Flow-Cytometric ParametersPET Imaging of Non-Small-Cell Lung Carcinoma with Carbon-11-Methionine:

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