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Germline mutations of the MYH gene in Japanese patients with multiple colorectal adenomas
Mutation Research 578 (2005) 430–433
Short communication
Germline mutations of the MYH gene in Japanese patients with multiple colorectal adenomas Michiko Miyaki a,∗ , Takeru Iijima a,e , Tatsuro Yamaguchi a,b , Tsunekazu Hishima c , Kazuo Tamura d , Joji Utsunomiya d , Takeo Mori b a
Hereditary Tumor Research Project, Tokyo Metropolitan Komagome Hospital, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8677, Japan b Department of Surgery, Tokyo Metropolitan Komagome Hospital, Tokyo 113-8677, Japan c Department of Pathology, Tokyo Metropolitan Komagome Hospital, Tokyo 113-8677, Japan d Institute for Advanced Medical Sciences, Hyogo College of Medicine, Nishinomiya 663-8501, Japan e Institute of Molecular Oncology, Showa University, Tokyo 142-8666, Japan Received 17 November 2004; received in revised form 7 January 2005; accepted 11 January 2005 Available online 10 May 2005
Abstract Germline mutations of the MYH gene have been revealed to associate with the recessive inheritance of multiple colorectal adenomas in Caucasian population. However, MYH mutations in Japanese patients have not yet been clarified. In an assessment of MYH mutations, we examined 35 Japanese patients with multiple colorectal adenomas who had neither dominant inheritance of colorectal tumors, nor germline APC mutations. One patient had a homozygous biallelic MYH mutation, R231C and three independent patients had monoallelic MYH mutations at a splice-site on exon 11 (IVS10-2 A to G). These four patients had 21 to around 100 colorectal adenomas and 1–3 synchronous colorectal carcinomas. The most common mutations in Caucasian patients, Y165C and G382D, were not detected in our Japanese cases. The MYH mutations detected in Japanese patients were novel and different from those detected among Caucasian, Indian and Pakistani patients, which suggests the existence of ethnic differentiation in MYH mutations. © 2005 Elsevier B.V. All rights reserved. Keywords: MYH; Germline mutation; Multiple colorectal adenomas; Japanese patients
1. Introduction A typical colorectal polyposis is familial adenomatous polyposis (FAP) (MIM 175100), an autosomal ∗ Corresponding author. Tel.: +81 3 3823 2101x4425; fax: +81 3 3823 5433. E-mail address: [email protected] (M. Miyaki).
0027-5107/$ – see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.mrfmmm.2005.01.017
dominant disorder, which is predisposed to hundreds or even thousands of colorectal adenomatous polyps and has a high risk of colorectal cancer at a young age. FAP cases with a small number of polyps are classified as attenuated FAP (AFAP). Both types of FAP are caused by germline mutations in the APC gene, and analyses of APC gene have been the major method for the determination of patients with multiple colorectal
M. Miyaki et al. / Mutation Research 578 (2005) 430–433
adenomas. To date, more than 800 germline APC mutations have been identified in FAP families. We also have identified 90 germline APC mutations in Japanese FAP families. However, there are a considerable number of cases without detectable APC mutations. Such cases include those without any obvious vertical transmission of adenomas, and therefore, are assumed to be FAP or AFAP patients arising as new (de novo) mutants. Recently, the base excision repair gene MYH has been revealed as being the causative gene of multiple colorectal adenomas (MIM 608456), which is an autosomal recessive disorder with a high risk of colorectal cancer. Biallelic (homozygous or compound heterozygous) and monoallelic MYH germline mutations have been detected in Caucasian patients with multiple colorectal adenomas having 3–100 polyps [1–6]. Similar mutations have also been detected in patients who appear to be classic FAP, having 100–1000 polyps [4,6,7]. Two MYH mutations, Y165C and G382D, have been found to be the most common in Caucasian patients [3,4,6,7], while E466X was detected in Indian, and Y90X in Pakistani patients [2]. However, MYH mutations in Japanese patients have not yet been clarified. To assess the characteristics of MYH mutations in Japanese patients, we selected patients who were assumed to fall into the category of multiple colorectal adenomas. Among these patients we found novel germline MYH mutations that are different from those detected in Caucasians.
2. Materials and methods We selected 35 patients who were assumed to fall into the category of multiple colorectal adenomas. Peripheral blood or normal tissue samples and family history information were obtained with informed consent, and the analysis of samples was approved by the Komagome Hospital Review Committee. Genomic DNA was extracted from peripheral blood or normal tissue using proteinase K–phenol–chloroform. DNA samples were analyzed for exons 1–16 and the exon–intron boundaries of the MYH gene by the PCR–SSCP. The PCR reaction mixture (6 l) contained 300 ng of genomic DNA, each 0.2 M primer, 25 M each of four dNTPs, 1× PCR buffer, Taq polymerase and [␣-32 P]dCTP. PCR primers were the same as those
431
previously reported [1]. PCR was performed under the following conditions: 5 min at 97 ◦ C once, 1 min at 94 ◦ C, 1 min at 58 ◦ C and 1 min at 72 ◦ C for 35 cycles and 10 min at 72 ◦ C. The products were diluted fivefold with formamide dye solution, followed by heating for 5 min at 80 ◦ C. For SSCP analysis, 2 l of each diluted sample was applied to 5% polyacrylamide gel containing 5% glycerol, electrophoresed and gel was exposed to X-ray film. Abnormal single-strand DNA fragments were eluted from the SSCP gel, amplified by asymmetrical PCR, and then sequenced by the dideoxy chain-termination method as previously described [8]. Homozygous mutations were confirmed by the absence of normal bands in the SSCP analysis, and the presence of two alleles in the PCR analysis using D1S2677.
3. Results and discussion We selected 35 patients who were assumed to fall into the category of multiple colorectal adenomas on the basis of the following criteria: (i) without any vertical transmission of colorectal adenomas or carcinomas, (ii) with a comparatively small number of adenomas, (iii) with synchronous colorectal carcinomas and (iv) without a detectable germline APC mutation. A biallelic germline MYH mutation was detected in patient PLK358 (41-year-old male). The mutation was homozygous for the exon 9 missense mutation, CGT (Arg) to TGT (Cys) at codon 231 (891 C to T). The homozygosity was confirmed by the absence of normal bands in the SSCP analysis, and the presence of two alleles in the PCR analysis using D1S2677 (Fig. 1a). This mutation was not detected in 80 controls without colorectal adenomas. This patient had more than 100 polyps, which were tubular adenomas with moderate to severe dysplasia, and a synchronous moderately differentiated adenocarcinoma (Dukes’A) of the rectum. There was no vertical transmission of colorectal adenomas or carcinomas, but a sibling developed lung cancer at the age of 39. Monoallelic germline MYH mutations were detected in three other independent patients, PLK68, PLK174 and PLK285. The mutation in these three patients was a splice-site variant, IVS102 A to G (892-2 A to G) (Fig. 1b), which was assumed to form aberrant mRNA. This mutation was not detected in the 80 controls. Patient PLK68 (20-yearold male) had approximately 50 colorectal polyps and
432
M. Miyaki et al. / Mutation Research 578 (2005) 430–433
Fig. 1. Examples of germline mutation of MYH gene. (a) SSCP pattern and sequence in the case of homozygous biallelic mutation. (b) SSCP pattern and sequence in the case of heterozygous monoallelic mutation.
a synchronous mucinous carcinoma of the ascending colon. He also had gastric polyps, an epithelial cyst and mandibular osteoma, but no vertical transmission of colorectal tumors was recognized. Patient PLK174 (50year-old male), without any family history, had more than 100 colorectal polyps, a carcinoma of the transverse colon and multiple gastric polyps. One of large colorectal polyps of this case had a K-ras mutation of GGT to TGT at codon 12, and two polyps had somatic APC mutation of GAA to TAA at codon 1306. Patient PLK285 (52-year-old male) had 21 colorectal adenomas with moderate to severe atypia and 3 synchronous well-differentiated adenocarcinomas, with 1 having developed in the ascending colon and 2 in the descending colon. His parents had no colorectal tumors, but one sibling had colorectal polyps. It has been reported that the hMSH6-interacting domain was located to the region that include codons 232–254 of hMYH [9]. The mutation at codon 231 detected in patient PLK358 appears to affect the interaction between hMYH and hMSH6, since codon 231 is adjacent to codon 232. Accordingly, the biallelic mutation of CGT (Arg) to TGT (Cys) at codon 231 is assumed to be pathogenic. With respect to the splice-
site variant, IVS 10-2 A to G, characteristics of this mutation have recently been studied in detail [10]. The authors reported that RT-PCR analysis using primers located in exons 10 and 12 revealed an mRNA transcript retaining the full intron 10 sequence, leading to generation of the premature stop codon within intron 10. Consequently, the IVS 10-2 G allele encodes a truncated MYH protein without a nuclear localization signal (NLS) and the binding sites for APE1 and PCNA proteins. These findings suggested the IVS 10-2 A to G variant is pathogenic. The present study revealed 1 (3%) biallelic and 3 (9%) monoallelic germline MYH mutations among 35 Japanese patients with multiple colorectal adenomas (Table 1), although the pathogenic significance of the monoallelic mutations remains to be clarified. Since these heterozygotes had no vertical transmission of colorectal tumors, it is possible that second MYH mutations may be present but were not detected. Alternatively, it remains possible that monoallelic MYH mutations may be one of the mild causative events [11] in multiple colorectal adenomas. It is also possible that monoallelic mutation, IVS 10-2 A to G, detected in our three patients may function as a
M. Miyaki et al. / Mutation Research 578 (2005) 430–433
433
Table 1 Germline MYH mutations in Japanese patients with multiple colorectal adenomas Patient
First MYH mutation
Second MYH mutation
Sex
Age (years)
Colorectal polyp
Colorectal carcinoma
Vertical transmission of colorectal tumor
PLK358 PLK68 PLK174 PLK285
R231C IVS10-2 A > G IVS10-2 A > G IVS10-2 A > G
R231C ND ND ND
Male Male Male Male
41 20 50 52
>100 50 >100 21
1 1 1 3
No No No No
ND: not detected.
dominant-negative, since three polyps from one of these patients showed somatic G to T mutations of K-ras and APC genes which have been reported to be specific to colorectal tumors among MYH-associated patients [1,2,12]. Such monoallelic MYH mutations have also been reported among patients with multiple adenomas and classic adenomatous polyposis [4,6]. In the case of Caucasian patients, two common MYH mutations, Y165C and G382D, have been detected in more than 80% of all cases [3,4,7]. However, these mutations were not detected in our Japanese cases. The MYH mutations detected in Japanese patients are novel and different from those reported in Caucasian, Indian and Pakistani patients. These results suggest the existence of ethnic differentiation in MYH mutations.
[6]
[7]
[8]
References [1] N. Al-Tassan, N.H. Chmiel, J. Maynard, N. Fleming, A.L. Livingston, G.T. Williams, A.K. Hodges, D.R. Davies, S.S. David, J.R. Sampson, J.P. Cheadle, Inherited variants of MYH associated with somatic G:C > T:A mutations in colorectal tumors, Nat. Genet. 30 (2002) 227–232. [2] S. Jones, P. Emmerson, J. Maynard, J.M. Best, S. Jordan, G.T. Williams, J.R. Sampson, J.P. Cheadle, Biallelic germline mutations in MYH predispose to multiple colorectal adenoma and somatic G:C > T:A mutations, Hum. Mol. Genet. 11 (2002) 2961–2967. [3] J.R. Sampson, S. Dolwani, S. Jones, D. Eccles, A. Ellis, D.G. Evans, I. Frayling, S. Jordan, E.R. Maher, T. Mak, J. Maynard, F. Pigatto, J. Shaw, J.P. Cheadle, Autosomal recessive colorectal adenomatous polyposis due to inherited mutations of MYH, Lancet 362 (2003) 39–41. [4] O.M. Sieber, L. Lipton, M. Crabtree, K. Heinimann, P. Fidalgo, R.K.S. Phillips, M.-L. Bisgaard, T. Orntoft, L.A. Aaltonen, S.V. Hodgson, H.J.W. Thomas, I.P.M. Tomlinson, Multiple colorectal adenomas, classic adenomatous polyposis, and germ-line mutations in MYH, N. Engl. J. Med. 348 (2003) 791–799. [5] S. Enholm, T. Hienonen, A. Suomalainen, L. Lipton, I. Tomlinson, V. Karja, M. Eskelinen, J.-P. Mecklin, A. Karhu, H.J.
[9]
[10]
[11]
[12]
Jarvinen, L.A. Aaltonen, Proportion and phenotype of MYHassociated colorectal neoplasia in a population-based series of Finnish colorectal cancer patients, Am. J. Pathol. 163 (2003) 827–832. L. Wang, L.M. Baudhuin, L.A. Boardman, K.L. Steenblock, G.M. Petersen, K.C. Halling, A.J. French, R.A. Johnson, L.J. Burgart, K. Rabe, N.M. Lindor, S.N. Thibodeau, MYH mutations in patients with attenuated and classic polyposis and with young-onset colorectal cancer without polyps, Gastroenterology 127 (2004) 9–16. V. Gismondi, M. Meta, L. Bonelli, P. Radice, P. Sala, L. Bertario, A. Viel, M. Fornasarig, A. Arrigoni, M. Gentile, M. Ponz-deLeon, L. Anselmi, C. Mareni, P. Bruzzi, L. Varesco, Prevalence of the Y165C, G382D and 1395delGGA germline mutations of the MYH gene in Italian patients with adenomatous polyposis coli and colorectal adenomas, Int. J. Cancer 109 (2004) 680–684. M. Miyaki, M. Konishi, R. Kikuchi-Yanoshita, M. Enomoto, T. Igari, K. Tanaka, M. Muraoka, H. Takahashi, Y. Amada, M. Fukayama, Y. Maeda, T. Iwama, Y. Mishima, T. Mori, M. Koike, Characteristics of somatic mutation of the adenomatous polyposis coli gene in colorectal tumors, Cancer Res. 54 (1994) 3011–3020. Y. Gu, A. Parker, T.M. Wilson, H. Bai, D.-Y. Chang, A.-L. Lu, Human MutY homolog, a DNA glycosylase involved in base excision repair, physically and functionally interacts with mismatch repair proteins human MutS homolog 2/human MutS homolog 6, J. Biol. Chem. 277 (2002) 11135–11142. H. Tao, K. Shimmura, T. Hanaoka, S. Natsukawa, K. Shaura, Y. Koizumi, Y. Kasuga, T. Ozawa, T. Tsujinaka, Z. Li, S. Yamaguchi, J. Yokota, H. Sugimura, S. Tsugane, A novel splice-site variant of the base excision repair gene MYH is associated with production of an aberrant mRNA transcript encoding a truncated MYH protein not localized in the nucleus, Carcinogenesis 25 (2004) 1859–1866. T. Kambara, V.L.J. Whitehall, K.J. Spring, M.A. Baker, S. Arnold, C.V.A. Wynter, N. Matsubara, N. Tanaka, J.P. Ypong, B.A. Leggett, J.R. Jass, Role of inherited defects of MYH in the development of sporadic colorectal cancer, Genes Chromosomes Cancer 40 (2004) 1–9. L. Lipton, S.E. Halford, V. Johnson, M.R. Novelli, A. Jones, C. Cummings, E. Barclay, O. Sieber, A. Sadat, M.-L. Bisgaard, S.V. Hodgson, L.A. Aaltonen, H.J.W. Thomas, I.P.M. Tomlinson, Carcinogenesis in MYH-associated polyposis follows a distinct genetic pathway, Cancer Res. 63 (2003) 7595–7599.
Short communication
Germline mutations of the MYH gene in Japanese patients with multiple colorectal adenomas Michiko Miyaki a,∗ , Takeru Iijima a,e , Tatsuro Yamaguchi a,b , Tsunekazu Hishima c , Kazuo Tamura d , Joji Utsunomiya d , Takeo Mori b a
Hereditary Tumor Research Project, Tokyo Metropolitan Komagome Hospital, 3-18-22 Honkomagome, Bunkyo-ku, Tokyo 113-8677, Japan b Department of Surgery, Tokyo Metropolitan Komagome Hospital, Tokyo 113-8677, Japan c Department of Pathology, Tokyo Metropolitan Komagome Hospital, Tokyo 113-8677, Japan d Institute for Advanced Medical Sciences, Hyogo College of Medicine, Nishinomiya 663-8501, Japan e Institute of Molecular Oncology, Showa University, Tokyo 142-8666, Japan Received 17 November 2004; received in revised form 7 January 2005; accepted 11 January 2005 Available online 10 May 2005
Abstract Germline mutations of the MYH gene have been revealed to associate with the recessive inheritance of multiple colorectal adenomas in Caucasian population. However, MYH mutations in Japanese patients have not yet been clarified. In an assessment of MYH mutations, we examined 35 Japanese patients with multiple colorectal adenomas who had neither dominant inheritance of colorectal tumors, nor germline APC mutations. One patient had a homozygous biallelic MYH mutation, R231C and three independent patients had monoallelic MYH mutations at a splice-site on exon 11 (IVS10-2 A to G). These four patients had 21 to around 100 colorectal adenomas and 1–3 synchronous colorectal carcinomas. The most common mutations in Caucasian patients, Y165C and G382D, were not detected in our Japanese cases. The MYH mutations detected in Japanese patients were novel and different from those detected among Caucasian, Indian and Pakistani patients, which suggests the existence of ethnic differentiation in MYH mutations. © 2005 Elsevier B.V. All rights reserved. Keywords: MYH; Germline mutation; Multiple colorectal adenomas; Japanese patients
1. Introduction A typical colorectal polyposis is familial adenomatous polyposis (FAP) (MIM 175100), an autosomal ∗ Corresponding author. Tel.: +81 3 3823 2101x4425; fax: +81 3 3823 5433. E-mail address: [email protected] (M. Miyaki).
0027-5107/$ – see front matter © 2005 Elsevier B.V. All rights reserved. doi:10.1016/j.mrfmmm.2005.01.017
dominant disorder, which is predisposed to hundreds or even thousands of colorectal adenomatous polyps and has a high risk of colorectal cancer at a young age. FAP cases with a small number of polyps are classified as attenuated FAP (AFAP). Both types of FAP are caused by germline mutations in the APC gene, and analyses of APC gene have been the major method for the determination of patients with multiple colorectal
M. Miyaki et al. / Mutation Research 578 (2005) 430–433
adenomas. To date, more than 800 germline APC mutations have been identified in FAP families. We also have identified 90 germline APC mutations in Japanese FAP families. However, there are a considerable number of cases without detectable APC mutations. Such cases include those without any obvious vertical transmission of adenomas, and therefore, are assumed to be FAP or AFAP patients arising as new (de novo) mutants. Recently, the base excision repair gene MYH has been revealed as being the causative gene of multiple colorectal adenomas (MIM 608456), which is an autosomal recessive disorder with a high risk of colorectal cancer. Biallelic (homozygous or compound heterozygous) and monoallelic MYH germline mutations have been detected in Caucasian patients with multiple colorectal adenomas having 3–100 polyps [1–6]. Similar mutations have also been detected in patients who appear to be classic FAP, having 100–1000 polyps [4,6,7]. Two MYH mutations, Y165C and G382D, have been found to be the most common in Caucasian patients [3,4,6,7], while E466X was detected in Indian, and Y90X in Pakistani patients [2]. However, MYH mutations in Japanese patients have not yet been clarified. To assess the characteristics of MYH mutations in Japanese patients, we selected patients who were assumed to fall into the category of multiple colorectal adenomas. Among these patients we found novel germline MYH mutations that are different from those detected in Caucasians.
2. Materials and methods We selected 35 patients who were assumed to fall into the category of multiple colorectal adenomas. Peripheral blood or normal tissue samples and family history information were obtained with informed consent, and the analysis of samples was approved by the Komagome Hospital Review Committee. Genomic DNA was extracted from peripheral blood or normal tissue using proteinase K–phenol–chloroform. DNA samples were analyzed for exons 1–16 and the exon–intron boundaries of the MYH gene by the PCR–SSCP. The PCR reaction mixture (6 l) contained 300 ng of genomic DNA, each 0.2 M primer, 25 M each of four dNTPs, 1× PCR buffer, Taq polymerase and [␣-32 P]dCTP. PCR primers were the same as those
431
previously reported [1]. PCR was performed under the following conditions: 5 min at 97 ◦ C once, 1 min at 94 ◦ C, 1 min at 58 ◦ C and 1 min at 72 ◦ C for 35 cycles and 10 min at 72 ◦ C. The products were diluted fivefold with formamide dye solution, followed by heating for 5 min at 80 ◦ C. For SSCP analysis, 2 l of each diluted sample was applied to 5% polyacrylamide gel containing 5% glycerol, electrophoresed and gel was exposed to X-ray film. Abnormal single-strand DNA fragments were eluted from the SSCP gel, amplified by asymmetrical PCR, and then sequenced by the dideoxy chain-termination method as previously described [8]. Homozygous mutations were confirmed by the absence of normal bands in the SSCP analysis, and the presence of two alleles in the PCR analysis using D1S2677.
3. Results and discussion We selected 35 patients who were assumed to fall into the category of multiple colorectal adenomas on the basis of the following criteria: (i) without any vertical transmission of colorectal adenomas or carcinomas, (ii) with a comparatively small number of adenomas, (iii) with synchronous colorectal carcinomas and (iv) without a detectable germline APC mutation. A biallelic germline MYH mutation was detected in patient PLK358 (41-year-old male). The mutation was homozygous for the exon 9 missense mutation, CGT (Arg) to TGT (Cys) at codon 231 (891 C to T). The homozygosity was confirmed by the absence of normal bands in the SSCP analysis, and the presence of two alleles in the PCR analysis using D1S2677 (Fig. 1a). This mutation was not detected in 80 controls without colorectal adenomas. This patient had more than 100 polyps, which were tubular adenomas with moderate to severe dysplasia, and a synchronous moderately differentiated adenocarcinoma (Dukes’A) of the rectum. There was no vertical transmission of colorectal adenomas or carcinomas, but a sibling developed lung cancer at the age of 39. Monoallelic germline MYH mutations were detected in three other independent patients, PLK68, PLK174 and PLK285. The mutation in these three patients was a splice-site variant, IVS102 A to G (892-2 A to G) (Fig. 1b), which was assumed to form aberrant mRNA. This mutation was not detected in the 80 controls. Patient PLK68 (20-yearold male) had approximately 50 colorectal polyps and
432
M. Miyaki et al. / Mutation Research 578 (2005) 430–433
Fig. 1. Examples of germline mutation of MYH gene. (a) SSCP pattern and sequence in the case of homozygous biallelic mutation. (b) SSCP pattern and sequence in the case of heterozygous monoallelic mutation.
a synchronous mucinous carcinoma of the ascending colon. He also had gastric polyps, an epithelial cyst and mandibular osteoma, but no vertical transmission of colorectal tumors was recognized. Patient PLK174 (50year-old male), without any family history, had more than 100 colorectal polyps, a carcinoma of the transverse colon and multiple gastric polyps. One of large colorectal polyps of this case had a K-ras mutation of GGT to TGT at codon 12, and two polyps had somatic APC mutation of GAA to TAA at codon 1306. Patient PLK285 (52-year-old male) had 21 colorectal adenomas with moderate to severe atypia and 3 synchronous well-differentiated adenocarcinomas, with 1 having developed in the ascending colon and 2 in the descending colon. His parents had no colorectal tumors, but one sibling had colorectal polyps. It has been reported that the hMSH6-interacting domain was located to the region that include codons 232–254 of hMYH [9]. The mutation at codon 231 detected in patient PLK358 appears to affect the interaction between hMYH and hMSH6, since codon 231 is adjacent to codon 232. Accordingly, the biallelic mutation of CGT (Arg) to TGT (Cys) at codon 231 is assumed to be pathogenic. With respect to the splice-
site variant, IVS 10-2 A to G, characteristics of this mutation have recently been studied in detail [10]. The authors reported that RT-PCR analysis using primers located in exons 10 and 12 revealed an mRNA transcript retaining the full intron 10 sequence, leading to generation of the premature stop codon within intron 10. Consequently, the IVS 10-2 G allele encodes a truncated MYH protein without a nuclear localization signal (NLS) and the binding sites for APE1 and PCNA proteins. These findings suggested the IVS 10-2 A to G variant is pathogenic. The present study revealed 1 (3%) biallelic and 3 (9%) monoallelic germline MYH mutations among 35 Japanese patients with multiple colorectal adenomas (Table 1), although the pathogenic significance of the monoallelic mutations remains to be clarified. Since these heterozygotes had no vertical transmission of colorectal tumors, it is possible that second MYH mutations may be present but were not detected. Alternatively, it remains possible that monoallelic MYH mutations may be one of the mild causative events [11] in multiple colorectal adenomas. It is also possible that monoallelic mutation, IVS 10-2 A to G, detected in our three patients may function as a
M. Miyaki et al. / Mutation Research 578 (2005) 430–433
433
Table 1 Germline MYH mutations in Japanese patients with multiple colorectal adenomas Patient
First MYH mutation
Second MYH mutation
Sex
Age (years)
Colorectal polyp
Colorectal carcinoma
Vertical transmission of colorectal tumor
PLK358 PLK68 PLK174 PLK285
R231C IVS10-2 A > G IVS10-2 A > G IVS10-2 A > G
R231C ND ND ND
Male Male Male Male
41 20 50 52
>100 50 >100 21
1 1 1 3
No No No No
ND: not detected.
dominant-negative, since three polyps from one of these patients showed somatic G to T mutations of K-ras and APC genes which have been reported to be specific to colorectal tumors among MYH-associated patients [1,2,12]. Such monoallelic MYH mutations have also been reported among patients with multiple adenomas and classic adenomatous polyposis [4,6]. In the case of Caucasian patients, two common MYH mutations, Y165C and G382D, have been detected in more than 80% of all cases [3,4,7]. However, these mutations were not detected in our Japanese cases. The MYH mutations detected in Japanese patients are novel and different from those reported in Caucasian, Indian and Pakistani patients. These results suggest the existence of ethnic differentiation in MYH mutations.
[6]
[7]
[8]
References [1] N. Al-Tassan, N.H. Chmiel, J. Maynard, N. Fleming, A.L. Livingston, G.T. Williams, A.K. Hodges, D.R. Davies, S.S. David, J.R. Sampson, J.P. Cheadle, Inherited variants of MYH associated with somatic G:C > T:A mutations in colorectal tumors, Nat. Genet. 30 (2002) 227–232. [2] S. Jones, P. Emmerson, J. Maynard, J.M. Best, S. Jordan, G.T. Williams, J.R. Sampson, J.P. Cheadle, Biallelic germline mutations in MYH predispose to multiple colorectal adenoma and somatic G:C > T:A mutations, Hum. Mol. Genet. 11 (2002) 2961–2967. [3] J.R. Sampson, S. Dolwani, S. Jones, D. Eccles, A. Ellis, D.G. Evans, I. Frayling, S. Jordan, E.R. Maher, T. Mak, J. Maynard, F. Pigatto, J. Shaw, J.P. Cheadle, Autosomal recessive colorectal adenomatous polyposis due to inherited mutations of MYH, Lancet 362 (2003) 39–41. [4] O.M. Sieber, L. Lipton, M. Crabtree, K. Heinimann, P. Fidalgo, R.K.S. Phillips, M.-L. Bisgaard, T. Orntoft, L.A. Aaltonen, S.V. Hodgson, H.J.W. Thomas, I.P.M. Tomlinson, Multiple colorectal adenomas, classic adenomatous polyposis, and germ-line mutations in MYH, N. Engl. J. Med. 348 (2003) 791–799. [5] S. Enholm, T. Hienonen, A. Suomalainen, L. Lipton, I. Tomlinson, V. Karja, M. Eskelinen, J.-P. Mecklin, A. Karhu, H.J.
[9]
[10]
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