HNPCC的突變篩查

1、HNPCC的突變篩查的突變篩查復(fù)旦大學(xué)附屬腫瘤醫(yī)院復(fù)旦大學(xué)附屬腫瘤醫(yī)院 大腸外科大腸外科劉方奇劉方奇2010-4-29Contents1.1.研究背景研究背景2.2.材料與方法材料與方法3.3.結(jié)果及討論結(jié)果及討論研究背景研究背景介紹 何為何為HNPCCHNPCC？常染色體顯性遺傳??；常染色體顯性遺傳病； 5-10%5-10%發(fā)生率；發(fā)生率；MMRMMR缺陷引起缺陷引起 如何研究？如何研究？臨床標(biāo)準(zhǔn)；選擇臨床標(biāo)準(zhǔn)；選擇MLH1MLH1和和MSH2MSH2基因篩查突變基因篩查突變 研究現(xiàn)狀研究現(xiàn)狀歐洲多，亞洲少歐洲多，亞洲少90%90%突變出現(xiàn)在突變出現(xiàn)在MLH1MLH1和和MSH2MSH2無熱

2、點突變無熱點突變研究背景介紹 臨床標(biāo)準(zhǔn)臨床標(biāo)準(zhǔn)Amsterdam I&IIAmsterdam I&II；BethesdaBethesda； 數(shù)據(jù)庫數(shù)據(jù)庫最權(quán)威最權(quán)威InSightInSight ( () ) 研究的意義是什么？研究的意義是什么？遺傳性疾病，早預(yù)防早治療遺傳性疾病，早預(yù)防早治療中國研究較少中國研究較少希望找到亞洲或中國的熱點突變希望找到亞洲或中國的熱點突變?yōu)橹袊呐R床檢測提供指導(dǎo)為中國的臨床檢測提供指導(dǎo)材料與方法材料與方法篩選98個疑似HNPCC病人根據(jù)臨床

3、標(biāo)準(zhǔn)：Bethesda, Amsterdam和復(fù)旦標(biāo)準(zhǔn)外周血樣提取PCR測序突變篩查突變判定新突變及熱點突變篩選文獻InSight數(shù)據(jù)庫結(jié)果與討論結(jié)果與討論臨床資料歸納與整理臨床資料歸納與整理1.突變結(jié)果討論突變結(jié)果討論2.臨床左半左半 右半右半64.7% vs. 35.3% 5222433202818154405101520253035404550 Ascending colon Hepatic flexure Transverse colon Splenic flexure Descending colon Sigmoid colon RectumAmsterdam criteriaBet

4、hesda criteriaAmsterdam I&II criteriaBethesda criteriaAlln(%)15(15.3%)83(84.7%)98(100%)Age(median, range) years48.1(35-67)47.6(19-82)47.7(19-82)Sex n(%) male12(80.0%)45(54.2%)57(58.2%) female3(20.0%)38(45.8%)41(41.8%)Location n(%) Ascending colon5(23.8%)20(20.4%)25(21.0%) Hepatic flexure2(9.5%)2

5、(2.0%)4(3.4%) Transverse colon2(9.5%)8(8.2%)10(8.4%) Splenic flexure2(9.5%)1(1.0%)3(2.5%) Descending colon4(19.0%)8(8.2%)12(10.1%) Sigmoid colon3(14.3%)15(15.3%)18(15.1%) Rectum3(14.3%)44(44.9%)47(39.5%)Multiple cancer synchronous tumours3(20.0%)3(3.6%)6(6.1%) metachronous tumour4(26.7%)17(20.5%)21(

6、21.4%)Pathology adenocarcinoma13(86.7%)69(83.1%)82(83.7%) mucinous carcinoma2(13.3%)11(13.2%)13(13.3%) Others0(0.0%)3(3.6%)3(3.1%)Family history155166同時異時腫瘤發(fā)生率高粘液腺癌10% 31 CRC happened, 11 were left side and 20 were right (64.5% vs. 35.5%), 20 related cancers happened, most of them were gastric cance

7、r and endometrial carcinoma, 5 each (25%). Family IDCRC numberCRC frequencyCancer numberCancer frequency MaleFemaleOnset age of CRCOnset age of cancerMultiple primary CRCMultiple primary tumorsRight siteLeft siteExtracolonic cancer (type) Mucinous carcinomaH94646314848225100H765555413737001400H86123

8、521474711113(1St, 1Es, 1E)1H88113330191900102(2L)0H1664545223636112300H167465732292922151(1E)0H224115723445102016(2St, 2E, 2Br)0H231112311474401012(1L, 1Br)0H236333412404001031(1E)1H245113612565001015(2St, 1Li, 1L, 1O)0 AC組組15例，突變例，突變4例例 如在腸外腫瘤中加入胃癌，符合如在腸外腫瘤中加入胃癌，符合30例，突例，突變變8例例 檢出率相同，避免遺漏檢出率相同，避免遺漏

9、 復(fù)旦推薦標(biāo)準(zhǔn)復(fù)旦推薦標(biāo)準(zhǔn)突變Family IDMutationClinical criteriaHaving been reported (times)GeneExonNucleotideConsequenceTypeH9MSH21c.23CTp.Thr8MetMissense AmsterdamYes (5)H76MLH12c.157del GAGG Frameshift AmsterdamNoH86MLH11c.-64GTUncertainBethesdaNoH88MSH27c.1168CTp.Phe390LeuMissenseBethesdaYes (34)H166MLH18c.655

10、AGp.Ile219ValMissense AmsterdamYes (224)H167MLH117c.1989GAAberrant splicingAmsterdamYes (2)H224MLH119c.2159insGFrameshiftBethesdaNoMSH27c.1168CTp.Phe390LeuMissenseYes (34)H231MSH27c.1168CTp.Phe390LeuMissenseBethesdaYes (34)H236MLH118c.2042CTp.Ala681ValMissenseBethesdaYes (1)H245MSH27c.1168CTp.Phe390

11、LeuMissenseBethesdaYes (34)該患者同時有兩個突變，且滿足復(fù)旦推薦標(biāo)準(zhǔn) 四個患者突變一致MSH2 c.1168CTInSight數(shù)據(jù)庫中該位點報道結(jié)果亞洲占絕大多數(shù)潛在亞洲人種熱點突變 or SNP4%9%5%5%45%32%FrenchAmericanAustralianKoreanJapaneseChinese參考文獻1.Lee S-C, Guo J-Y, Lim R et al. Clinical and molecular characteristics of hereditary non-polyposis colorectal cancer families

12、 in Southeast Asia. Clinical Genetics 2005: 68: 137-145.2.Landis SH, Murray T, Bolden S et al. Cancer statistics, 1999. CA Cancer J Clin 1999: 49: 8-31, 31.3.Lynch HT, Smyrk T. Hereditary nonpolyposis colorectal cancer (Lynch syndrome): An updated review. Cancer 1996: 78: 1149-1167.4.Ricciardiello L

13、, Boland C. Lynch syndrome (hereditary non-polyposis colorectal cancer): Current concepts and approaches to management. Current Gastroenterology Reports 2005: 7: 412-420.5.Lynch HT, Lynch JF, Lynch PM. Toward a consensus in molecular diagnosis of hereditary nonpolyposis colorectal cancer (Lynch synd

14、rome). J Natl Cancer Inst 2007: 99: 261-263.6.Lynch HT, de la Chapelle A. Hereditary Colorectal Cancer. The New England Journal of Medicine 2003: 348: 919-932.7.Vasen HFA, Watson P, Mecklin J-P et al. New clinical criteria for hereditary nonpolyposis colorectal cancer (HNPCC, Lynch syndrome) propose

15、d by the International Collaborative Group on HNPCC. Gastroenterology 1999: 116: 1453-1456.8.Lynch HT, de la Chapelle A. Genetic susceptibility to non-polyposis colorectal cancer. Journal of Medical Genetics 1999: 36: 801-818.9.Vasen HFA, Wijnen JT, Menko FH et al. Cancer risk in families with hereditary nonpolyposis colorectal cancer diagnosed by mutation analysis. Gastr