- Email: [email protected]
Mucosal expression of the fragile histidine triad in gastric carcinoma in relation to type, grade and stage of tumour
Oncology
161 Mucosal expression of the fragile bistidine hiad in gastic carcinoma in relation to type, grade and stageof hsmolu Rocm A,1 SchandlL,2 Z. Tulassay,Z MalfertheinerP.3 Eberl M.3 Bud&n G,l NardoneG.l lDepart!aent of Clinical and ExperimentalMedicine, ‘Fed&co II’ University, Naples, Italy Xkpartment of Internal Medicine, SemmelweisUniversity, Budapest,Hungary 3Deparbnentof Gastmentemlogy,Otto-van-Guericke Umversity, Magdeburg,Germany Diaztimento di Me&&a Clinica e Stientale B&kgmund: Gastric cawer is cons&ted by two bistomorphologicalentities(“intestinal” and “di!Tbse”) that differ in epidemiology,pathogenesis,clinical outcome and geneticprofile. Alterationsof the fragilehistidine triad (Fhit) genehave beenidentified in a vari&y of human malignanciessuggestingthat Fhit might be a tumor suppresscr gene.Aim: To investigateFhit protein expressionin human primary gastric carcinoma in relation to type, gradeand stageof hunour. Material andmethods: The study population cons&d of 137patients(100 malesand 37 females;mean age60.5 years) who bad undergonesurgery for gastric carcinoma. Gastric cancer was evaluatedaccording to Lauren’s classification. Fhif protein exprpression was evaluatedon gastic samplesof tumor or tumor-fw areasby immunobistcchemis~xyusing specific polyclonal antibodies(Zymed Laboratories). In a subgroup of 30 patients,Fbit mRNA expression was detected by nestedRT-PCR Results: Gastric cancer was “intestinal” in 99 casesand “diffuse” in 38 cases; well differentiatedin 25, moderatelydifferentmledm 35 andpoorly differentiated in 77; early stage in 6 (all intes~ types) andadvancedin 131.In tumoral areas,Fhit protein expressionwas significantly lower in diffuse, in poorly differentiatedand in advancedcan- than in intestinal, well differentiatedand early gastric cancers (p
163 Microsatellite instability andtarget genemutationsin adenomatouspolyps of patientswith a single iirstdegres memberaffected by colon cancer. Luigi Ricciardiello, Ajay Gael*, Wilma Mantovam#,Lorenm Fuccio, StefaniaFossi, Dong K. Chang*, Veronica Lunedei, Filippa A&mini, C. Richard B&ad*, France Bazmli and Enrico Roda Dipartimentodi Medic& Interna e Gasmentemlogia Universit& di Bologna, #Laboratorio Centdizzata PoIiclinico S. Orsala Bologna, *Cancer CenterUC San Diego Paliclinico S. Orsola Background: Colorectal cancers arise fmm pre-existing adenomatouspolyps. It is widely accepted that the APC zeneis the natekeeoerfor thecolon carcinwenesis and subsequentclonal expansion might lead to&osatell& ins&lily (MSI) or chmnms&al instability (C-w. MS1 is the landmark phenomenonin hereditary non-palypasis colorectal syndrome (HNPCC) but can also be found in a-boutl5-20% of sporadic-colorectal&ncers, mostly due to hMLH1 promoter hypermetbylation. We have previously demonstratedthat fust degreemembersof a singlepatient having colon cancer have a doubledrisk for developingc&rectal adenomas.Aims: In this study we sought to evaluatethe 6-equencyof MSI and targetgenemutations in adenotnasof patientswith andwithout a single first degreemember affectedby colon cancer. Methods: A prelimii analysisW&Pperfomml on 44 polyps obtained fmm 44 patients. DNA was extracted by microdissecting the tumor and,separately,the swmunding normal tissue.A panel of five recotmnendedmarkers was usedto determineihe MSI status. Tumors ihat showed instabilitv in at leasttwo markers were classified as MSI-H, whereasotheaxwere classified as MSI-L (instability at one locus) or MSS (no instability). Microsatellites within key target genes(MBD4, TGF?RII, BAX, MSH3 andMSH6) were also examinedin all thesamples. Extracted DNA was PCR amplified. Fra@nentanalysiswas performedby an automatedsequencer. Family history was carefully obtainedby qualified personnel.Results: Of the 44 polyps, 5 were MS&H (11.3%), 14were MSI-L (31.8) andthe remainderswere MSS (43.2%). Moreover, 2 patients showed LOH (4.5%) and4 displayedboth MS&L andLOH (9%). Of the 5 MSI-H polyps, two had B somatic tieshift mutation of the MBD4 gene,oneof the MSH6 andone of BAX. None of tbe MSI-L, the MSILiLOH, the LOH andMSS polyps hadmutations at target genes.Four out of five of the patients with MSI-H polyps had apositive family history of colon cancer and 3 of them (75%) had also target genemutations. 9% of patienfswithpositive family histmy had MS&H. Conclusions: Almost 10%with a simplefamily history for CRC had MSI-H along with target genemutations (probably dueto defective DNA MMR functionisystem). The higher rate of MS&H and target gene mutations h patientswith a simplefamily history for CRC and bearingadenomasmight be the explanationfor the higher rate of polyps comparedthe generalpopulation.
162 Protein expression of tumor suppressor genep16 in gastric cancer A. Rocco,l L. SchandI,ZZ.Tulassay,Z P. Malfertbeiner.3M. Ebert,3G. Bud&m,1 G. Nxdone,l lDepmtment of Clinical and ExperimentalMedicine, Gastmentemlogy Unit, University “Federiw IP . NaplesI Italy 211Deparbmd of Internal Medicine SemmelweisUniversity, Budapest Hungary 3Depatment of Gaatmeniemlogy,Dttc-vowGuticke-University, Magdeburg,Germany Dip Medicina Clinica e SpedmentaleFedaiw II Background: Mounting evidence points to a close relationshipbetweentumorigenesis and alterationsof the proteins regulatingthe cell cycle. pl6 is a cell-cycle regulatay protein acting 85a &in-dependent kinwe inhibitor. Becauseof its antipmlifemtive effect, pI6 is thought to be B tumor-suppressor gene.Delefions, mutationsor functional inactivation involving p16 are frequent in most bumm malignancies.Gastic cancer consis4sof two bistomoq$~ologicalentities (“‘intestinal” and“diffuse”) chsrsctnizcd by different genetic and pathogen& pathways. Aim: To evaluate~16 pmtein expression in gastric cancer in relation to histotypc and differentiation of tumor. idated and methods:The study population consistedof 70 patients(M/F 50120;mean age 60.4 years) who bad undergonesurgery for gastric cancer. The expression of thep16 protein was evaltied in tumor and in tumor-&e ureasby bmmmobistochemistry with specific polyclonal antibodies(pbanningen). Results: Gastric cancarwas “intestinal” in 52 patientsand ‘diffuse” in 18. In gastric cancer tissues, p16 was overexpressedin both intestinal anddi&se gz&ic cancer types. However, the Iwde of inummoreactivitv was inversely relatedto the made of tumoral differentiation~Jntumor-f&e areas~16 &pression xv& weak and loc&zed predominantly in the astral glands.Conclusions: The expression ofpI appearsto be a common event in both the intestinal anddiEuse type of gastric cancer. ‘IXe low expression found in poorly differentiated tumors suggeststhat the pl6 protein might be considereda prognostic factor in gastric cancer
Mad-l is the Exclusive JC Vim Strain Presentin tbe Human Colon and Deletesa 98-bp Sequence fmm the Transcriptional Control Region in Colon Cancers Luigi ficciardiello, Dong K. Cbang,Luigi Laghi, Ajay Gc-sl, Christina L. CbangandC. Richard Boland Fmm the Departmentof Medicine and Cancer Center,University of Callfonda, San Diego Policlinico S. @sola Background: JC vims (JCV). along with other membersof the polyomaviras family, encodesa class of highly conserved proteins,T antigas, that are capableof inducing aneuplaidyin cultured cells. We have previously isolatedT-antigen DNA variants of JCV fmm both colon cancers andtie corresponding non-neoplasticgastmintestimdtissues, raising new questionscmthe role of JCV in the developmentof cbmmowmal instability of the colon. Basedcmtbe sequenceof the transaipticmal control region (TCR), JCV can be classified as archme or tandemrepeatvariants. Among the latter, Mad-l, the prototype vims fbxt isolatedfrom a patientwith Progressive Multifocal Lwkoencephalopathy (PML), is characterizedby lacking the 23- and 66-bp sequences that are presentin the arc&Q+ aadby duplicationof a 98-bp sequence.Aim: In this study, we evaluateddifferences in tbe TCR of JCV isolated fmm colon cancers and non-neoplastic epithelium. Methods: To chamctexizeJCV vatiaou, we first treatedeight pairs of tiNA samples from don cancers andnon-cancemus tissuewith topoisomemseI andthen amplified andcloned the JCV TCR Results: We obtained 285 recombinantcloneafmm the JCV TCR 157fmm nonneoplasticsamplesand 128 from colon cancers. Of theseclones,262 spannedthe length of the JCV Mad-l TCR: 99.3% fmm non-nmolwtic sanmlesand 82.8% fmm colon cancers. In sauencine 54 clones in both directions, we did not lind archJCV either in the non-neoplastictissue or & the cancer samples.From all colon cancer tissues, I8 cloneshad a deletion of one98-bp tan&m repeat. This deleted strain was not detectedin any of the non-nmplasti~tissues (14% vs. O%,c2= 23.6, P
A79
161 Mucosal expression of the fragile bistidine hiad in gastic carcinoma in relation to type, grade and stageof hsmolu Rocm A,1 SchandlL,2 Z. Tulassay,Z MalfertheinerP.3 Eberl M.3 Bud&n G,l NardoneG.l lDepart!aent of Clinical and ExperimentalMedicine, ‘Fed&co II’ University, Naples, Italy Xkpartment of Internal Medicine, SemmelweisUniversity, Budapest,Hungary 3Deparbnentof Gastmentemlogy,Otto-van-Guericke Umversity, Magdeburg,Germany Diaztimento di Me&&a Clinica e Stientale B&kgmund: Gastric cawer is cons&ted by two bistomorphologicalentities(“intestinal” and “di!Tbse”) that differ in epidemiology,pathogenesis,clinical outcome and geneticprofile. Alterationsof the fragilehistidine triad (Fhit) genehave beenidentified in a vari&y of human malignanciessuggestingthat Fhit might be a tumor suppresscr gene.Aim: To investigateFhit protein expressionin human primary gastric carcinoma in relation to type, gradeand stageof hunour. Material andmethods: The study population cons&d of 137patients(100 malesand 37 females;mean age60.5 years) who bad undergonesurgery for gastric carcinoma. Gastric cancer was evaluatedaccording to Lauren’s classification. Fhif protein exprpression was evaluatedon gastic samplesof tumor or tumor-fw areasby immunobistcchemis~xyusing specific polyclonal antibodies(Zymed Laboratories). In a subgroup of 30 patients,Fbit mRNA expression was detected by nestedRT-PCR Results: Gastric cancer was “intestinal” in 99 casesand “diffuse” in 38 cases; well differentiatedin 25, moderatelydifferentmledm 35 andpoorly differentiated in 77; early stage in 6 (all intes~ types) andadvancedin 131.In tumoral areas,Fhit protein expressionwas significantly lower in diffuse, in poorly differentiatedand in advancedcan- than in intestinal, well differentiatedand early gastric cancers (p
163 Microsatellite instability andtarget genemutationsin adenomatouspolyps of patientswith a single iirstdegres memberaffected by colon cancer. Luigi Ricciardiello, Ajay Gael*, Wilma Mantovam#,Lorenm Fuccio, StefaniaFossi, Dong K. Chang*, Veronica Lunedei, Filippa A&mini, C. Richard B&ad*, France Bazmli and Enrico Roda Dipartimentodi Medic& Interna e Gasmentemlogia Universit& di Bologna, #Laboratorio Centdizzata PoIiclinico S. Orsala Bologna, *Cancer CenterUC San Diego Paliclinico S. Orsola Background: Colorectal cancers arise fmm pre-existing adenomatouspolyps. It is widely accepted that the APC zeneis the natekeeoerfor thecolon carcinwenesis and subsequentclonal expansion might lead to&osatell& ins&lily (MSI) or chmnms&al instability (C-w. MS1 is the landmark phenomenonin hereditary non-palypasis colorectal syndrome (HNPCC) but can also be found in a-boutl5-20% of sporadic-colorectal&ncers, mostly due to hMLH1 promoter hypermetbylation. We have previously demonstratedthat fust degreemembersof a singlepatient having colon cancer have a doubledrisk for developingc&rectal adenomas.Aims: In this study we sought to evaluatethe 6-equencyof MSI and targetgenemutations in adenotnasof patientswith andwithout a single first degreemember affectedby colon cancer. Methods: A prelimii analysisW&Pperfomml on 44 polyps obtained fmm 44 patients. DNA was extracted by microdissecting the tumor and,separately,the swmunding normal tissue.A panel of five recotmnendedmarkers was usedto determineihe MSI status. Tumors ihat showed instabilitv in at leasttwo markers were classified as MSI-H, whereasotheaxwere classified as MSI-L (instability at one locus) or MSS (no instability). Microsatellites within key target genes(MBD4, TGF?RII, BAX, MSH3 andMSH6) were also examinedin all thesamples. Extracted DNA was PCR amplified. Fra@nentanalysiswas performedby an automatedsequencer. Family history was carefully obtainedby qualified personnel.Results: Of the 44 polyps, 5 were MS&H (11.3%), 14were MSI-L (31.8) andthe remainderswere MSS (43.2%). Moreover, 2 patients showed LOH (4.5%) and4 displayedboth MS&L andLOH (9%). Of the 5 MSI-H polyps, two had B somatic tieshift mutation of the MBD4 gene,oneof the MSH6 andone of BAX. None of tbe MSI-L, the MSILiLOH, the LOH andMSS polyps hadmutations at target genes.Four out of five of the patients with MSI-H polyps had apositive family history of colon cancer and 3 of them (75%) had also target genemutations. 9% of patienfswithpositive family histmy had MS&H. Conclusions: Almost 10%with a simplefamily history for CRC had MSI-H along with target genemutations (probably dueto defective DNA MMR functionisystem). The higher rate of MS&H and target gene mutations h patientswith a simplefamily history for CRC and bearingadenomasmight be the explanationfor the higher rate of polyps comparedthe generalpopulation.
162 Protein expression of tumor suppressor genep16 in gastric cancer A. Rocco,l L. SchandI,ZZ.Tulassay,Z P. Malfertbeiner.3M. Ebert,3G. Bud&m,1 G. Nxdone,l lDepmtment of Clinical and ExperimentalMedicine, Gastmentemlogy Unit, University “Federiw IP . NaplesI Italy 211Deparbmd of Internal Medicine SemmelweisUniversity, Budapest Hungary 3Depatment of Gaatmeniemlogy,Dttc-vowGuticke-University, Magdeburg,Germany Dip Medicina Clinica e SpedmentaleFedaiw II Background: Mounting evidence points to a close relationshipbetweentumorigenesis and alterationsof the proteins regulatingthe cell cycle. pl6 is a cell-cycle regulatay protein acting 85a &in-dependent kinwe inhibitor. Becauseof its antipmlifemtive effect, pI6 is thought to be B tumor-suppressor gene.Delefions, mutationsor functional inactivation involving p16 are frequent in most bumm malignancies.Gastic cancer consis4sof two bistomoq$~ologicalentities (“‘intestinal” and“diffuse”) chsrsctnizcd by different genetic and pathogen& pathways. Aim: To evaluate~16 pmtein expression in gastric cancer in relation to histotypc and differentiation of tumor. idated and methods:The study population consistedof 70 patients(M/F 50120;mean age 60.4 years) who bad undergonesurgery for gastric cancer. The expression of thep16 protein was evaltied in tumor and in tumor-&e ureasby bmmmobistochemistry with specific polyclonal antibodies(pbanningen). Results: Gastric cancarwas “intestinal” in 52 patientsand ‘diffuse” in 18. In gastric cancer tissues, p16 was overexpressedin both intestinal anddi&se gz&ic cancer types. However, the Iwde of inummoreactivitv was inversely relatedto the made of tumoral differentiation~Jntumor-f&e areas~16 &pression xv& weak and loc&zed predominantly in the astral glands.Conclusions: The expression ofpI appearsto be a common event in both the intestinal anddiEuse type of gastric cancer. ‘IXe low expression found in poorly differentiated tumors suggeststhat the pl6 protein might be considereda prognostic factor in gastric cancer
Mad-l is the Exclusive JC Vim Strain Presentin tbe Human Colon and Deletesa 98-bp Sequence fmm the Transcriptional Control Region in Colon Cancers Luigi ficciardiello, Dong K. Cbang,Luigi Laghi, Ajay Gc-sl, Christina L. CbangandC. Richard Boland Fmm the Departmentof Medicine and Cancer Center,University of Callfonda, San Diego Policlinico S. @sola Background: JC vims (JCV). along with other membersof the polyomaviras family, encodesa class of highly conserved proteins,T antigas, that are capableof inducing aneuplaidyin cultured cells. We have previously isolatedT-antigen DNA variants of JCV fmm both colon cancers andtie corresponding non-neoplasticgastmintestimdtissues, raising new questionscmthe role of JCV in the developmentof cbmmowmal instability of the colon. Basedcmtbe sequenceof the transaipticmal control region (TCR), JCV can be classified as archme or tandemrepeatvariants. Among the latter, Mad-l, the prototype vims fbxt isolatedfrom a patientwith Progressive Multifocal Lwkoencephalopathy (PML), is characterizedby lacking the 23- and 66-bp sequences that are presentin the arc&Q+ aadby duplicationof a 98-bp sequence.Aim: In this study, we evaluateddifferences in tbe TCR of JCV isolated fmm colon cancers and non-neoplastic epithelium. Methods: To chamctexizeJCV vatiaou, we first treatedeight pairs of tiNA samples from don cancers andnon-cancemus tissuewith topoisomemseI andthen amplified andcloned the JCV TCR Results: We obtained 285 recombinantcloneafmm the JCV TCR 157fmm nonneoplasticsamplesand 128 from colon cancers. Of theseclones,262 spannedthe length of the JCV Mad-l TCR: 99.3% fmm non-nmolwtic sanmlesand 82.8% fmm colon cancers. In sauencine 54 clones in both directions, we did not lind archJCV either in the non-neoplastictissue or & the cancer samples.From all colon cancer tissues, I8 cloneshad a deletion of one98-bp tan&m repeat. This deleted strain was not detectedin any of the non-nmplasti~tissues (14% vs. O%,c2= 23.6, P
A79