аЯрЁБс>ўџ -/ўџџџ,џџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџьЅС@ №ПђbjbjУ.У. . ЁDЁDђџџџџџџˆ№№№№№№№hhhh,” &цЌЌЌЌЌЌЌЌЅЇЇЇЇЇЇ$ R^:Ы№cЌЌccЫ№№ЌЌр///cš№Ќ№ЌЅ/cЅ//Vщ@№№AЌ  ™x@ZЦh§") ‰і0&5 ˜˜A№№№№˜№AHЌ>ъ,/$:)ЌЌЌЫЫdh% hLoss of imprinting of IGF2 and H19; promoter usage of IGF2; loss of heterozigosity of IGF2R and Helicobacter pylori infection in laryngeal squamous cell carcinoma (LSCC) Ivana Grbesa, Koraljka Gall-Troselj Laboratory of Molecular Pathology, Division of Molecular Medicine, Rudjer Boskovic Institute, Bijenicka cesta 54, 10002 Zagreb, Croatia Abstract The imprinted gene IGF2 codes for the mitogenic peptide, comprising 67 amino acids and contributing to tumor growth through its autocrine or endocrine effects. Its expression, directed from four different promoters, is regulated on the level of DNA methylation, since it is reciprocally imprinted with H19. IGF2 biallelic expression is known as loss of imprinting (LOI), which physiologically occurs in prenatal liver and chondrocytes, from the P1 promoter. IGF2/H19 LOI, IGF2-R loss of heterozygosity (LOH) analysis and presence of H. pylori were tested on 36 squamous laryngeal carcinomas (LSCC), in addition to IGF2 promoter usage analysis in all non-tumorous and tumorous laryngeal tissue that expressed IGF2. For imprinting analyses a RFLP based method was used; IGF2-R LOH was detected by PCR followed by polyacrilamide gel electrophoresis/silver staining; H. pylori presence was detected by nested PCR and promoter usage was analysed by RT PCR with promoter specific primers. The informativity for loss of imprinting (LOI) analyses was: 44% (16/36) for the IGF2 and 53% (19/36) for the H19. Biallelic expression of IGF2 was observed in 27% (4/15) of non-tumorous laryngeal tissues and in 40% (6/15) of LSCC tumors. H19 LOI was observed in 24% (4/17) of LSCC tumors. One tumor sample showed LOH of both IGF2 and H19 coupled with monoallelic expression of these genes. For IGF2R, 69% (25/36) of samples were heterozygous and LOH was detected in 12% (3/25) tumors. H. pylori was found in 16% (13/82) of carcinoma tissues. P1 promoter was used in 4/15 (26%) non-tumors (two with IGF2 LOI) and in 8/15 (53%) LSCC tumors (four with IGF2 LOI). We conclude that IGF2 and H19 imprinting is disturbed in LSCC and IGF2 promoter usage is highly variable in malignant and adjacent normal laryngeal tissue. The presence of H. pylori had no effect on IGF2/H19 imprinting. In addition, the P1 promoter usage is not responsible for IGF2 LOI. #6:V\`sЋЮЯаX d v {  Ї З Ч h i ’ • — › е ж . 1 3 6 ; < јяјяјяјяјяјцлцајРЎРžРžРžРЎРЎРžР‘p‘Рh`Ў56CJKHOJQJ"h`Ўh`Ў56CJKHOJQJh`Ў5CJKHOJQJhСVOhсxЦ5CJKHOJQJ"hСVOhАbD56CJKHOJQJhСVOhАbD5CJKHOJQJhАbD56OJQJhАbD5H*OJQJhАbD5OJQJhАbD6OJQJhАbDOJQJ%ЊЋЯаX Y Z c d ёђїѕьцѕѕѕѕѕзв$a$$„h*$7$8$H$`„ha$ ЦJ $ ЦJ a$$ & Fa$ ђў< @ C [ ^ ` h l y ‚ ƒ ” В Й Ъ Ю ь  ' , . E G d j Т юобобобоюоСБбБŸб’б€бrdrTFhСVOh}}“5CJOJQJhСVOh}}“56CJOJQJhСVOhМv-5CJOJQJhСVOhАbD5CJOJQJ"h`Ўh`Ў56CJKHOJQJh}}“5CJKHOJQJ"h}}“h`Ў56CJKHOJQJhСVOhАbD5CJKHOJQJhСVOhСVO5CJKHOJQJh`Ў5CJKHOJQJhСVOh`Ў5CJKHOJQJ"hСVOh`Ў56CJKHOJQJТ У Ь ё ѕ ї   8 9 : ; Œ  Љ Ќ Ў Ц Ъ *-]єфжєШКЌШžШ‚r‚r‚bbO$hСVOhѕB5B*CJOJQJphhСVOhѕB56CJOJQJhСVOhСVO56CJOJQJhСVOhСVO5CJOJQJhСVOhѕB5CJOJQJhСVOhМv-5CJOJQJhСVOho_5CJOJQJhСVOhф|5CJOJQJhСVOhАbD5CJOJQJhСVOh}}“5CJOJQJhСVOh}}“56CJOJQJh}}“5CJOJQJ…ŠЦЬ!*@Z‘–ХЩЮЯасыиыиыиХЕЇœ‰u‰u‰ZJ7$hСVOhАbD5B*CJOJQJphhСVOhАbD5CJKHOJQJ4hСVOhšRG5@ˆB*CJKHOJQJnH phtH 'hСVOh}}“56B*CJOJQJph$hСVOh}}“5B*CJOJQJphh}}“5CJOJQJhСVOhѕB5CJOJQJhСVOhѕB56CJOJQJ$hСVOhѕB5B* CJOJQJph€$hСVOhѕB5B*CJOJQJph'hСVOhѕB56B*CJOJQJphсхъэl|…—ŸЊЋЌцъ№ёђыиыиыиЪКЪКЪЏœиыиqhСVOhАbD5CJKHOJQJ4hСVOhАbD5@ˆB*CJKHOJQJnH phtH $hСVOh}}“5B*CJOJQJphh}}“5CJOJQJhСVOh}}“56CJOJQJhСVOh}}“5CJOJQJ$hСVOhАbD5B*CJOJQJph'hСVOhАbD56B*CJOJQJph,1hАа/ Ар=!А"А# $ %ААФАФ ФœT@ёџT Normal*$'5@ˆ CJKHOJQJ_HmH sH tH\@\ Heading 1$ & FЄ№Є<@&CJ OJQJ\^JaJ DA@ђџЁD Default Paragraph FontVi@ѓџГV  Table Normal :V і4ж4ж laі (k@єџС(No List H™@ђH АbD Balloon TextCJOJQJ^JaJђ џџџџЊЋЯаXYZcdёє0€€˜0€€˜0€€˜0€€˜0€€˜0€€˜0€€˜0€€˜0€€˜0€€˜0€€< Т сђ ђ ђ џџ-Ф2  oћ.Ф2Loћ/Ф2œ^ 0Ф2, ЙHHPєЮNWWє9*€urn:schemas-microsoft-com:office:smarttags€place€8*€urn:schemas-microsoft-com:office:smarttags€City€B*€urn:schemas-microsoft-com:office:smarttags€country-region€>*€urn:schemas-microsoft-com:office:smarttags €PersonName€ DRЋАБЗ".78=Š“фэ ?Lє$.ЎЯє33Ћа.78;<<C[^`hlmmvv‚ƒˆˆ“”ВЙЪ.ddТУ№ѕ99:ЮааlЌєєџџџџџџџџџЁc(ШƒNџџџџџџџџџ[TЗi:^к'џџџџџџџџџџ„„Ц^„`„џ„„Ц^„`„џ„„Ц^„`„џ„„Ц^„`„џ„„Ц^„`„џ„„Ц^„`„џ„„Ц^„`„џ„„Ц^„`„џ„„Ц^„`„„а„˜ўЦа^„а`„˜ўOJPJQJ^Jo(-„ „˜ўЦ ^„ `„˜ўOJQJ^Jo(‡hˆHo€„p„˜ўЦp^„p`„˜ўOJQJo(‡hˆHЇ№€„@ „˜ўЦ@ ^„@ `„˜ўOJQJo(‡hˆHЗ№€„„˜ўЦ^„`„˜ўOJQJ^Jo(‡hˆHo€„р„˜ўЦр^„р`„˜ўOJQJo(‡hˆHЇ№€„А„˜ўЦА^„А`„˜ўOJQJo(‡hˆHЗ№€„€„˜ўЦ€^„€`„˜ўOJQJ^Jo(‡hˆHo€„P„˜ўЦP^„P`„˜ўOJQJo(‡hˆHЇ№h„а„˜ўЦа^„а`„˜ўOJPJQJ^Jo(-h„ „˜ўЦ ^„ `„˜ўOJQJ^Jo(‡hˆHoh„p„˜ўЦp^„p`„˜ўOJQJo(‡hˆHЇ№h„@ „˜ўЦ@ ^„@ `„˜ўOJQJo(‡hˆHЗ№h„„˜ўЦ^„`„˜ўOJQJ^Jo(‡hˆHoh„р„˜ўЦр^„р`„˜ўOJQJo(‡hˆHЇ№h„А„˜ўЦА^„А`„˜ўOJQJo(‡hˆHЗ№h„€„˜ўЦ€^„€`„˜ўOJQJ^Jo(‡hˆHoh„P„˜ўЦP^„P`„˜ўOJQJo(‡hˆHЇ№Ёc([TЗiџџџџџџџџџџџџџџџџ х ѕBМv-e^;АbDšRGСVOљ+Po_ф|}}“`ЎсxЦcЮєџџўџ@€ёёdPёёђ`@џџUnknownџџџџџџџџџџџџG‡z €џTimes New Roman5€Symbol3& ‡z €џArial;AdvPS6F00A& ‡ŸArial Narrow5& ‡za€џTahoma?5 ‡z €џCourier New;€Wingdings"qŒ№аhŽ:ЄІŽ:ЄІk:ЄІUU!№ ДД24dюю3ƒQ№ппHP(№џ?фџџџџџџџџџџџџџџџџџџџџџсxЦџџ~Loss of imprinting of IGF2 and H19, loss of heterozigosity of IGF2R and Helicobacter pylori infection in laryngeal squamous ceIvanaIvana   ўџр…ŸђљOhЋ‘+'Гй0р˜ ,<HX ht  œ Ј ДРШаифLoss of imprinting of IGF2 and H19, loss of heterozigosity of IGF2R and Helicobacter pylori infection in laryngeal squamous ceossIvanaf vanNormal Ivana 2anMicrosoft Word 10.0@Œ†G@кМфZЦ@Tb9ZЦ@Tb9ZЦUўџеЭеœ.“—+,љЎ0h hp|„Œ” œЄЌД М GфirbюO Loss of imprinting of IGF2 and H19, loss of heterozigosity of IGF2R and Helicobacter pylori infection in laryngeal squamous ce Title ўџџџўџџџ !"#ўџџџ%&'()*+ўџџџ§џџџ.ўџџџўџџџўџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџRoot Entryџџџџџџџџ РFb‰@ZЦ0€1TableџџџџџџџџАWordDocumentџџџџџџџџ. SummaryInformation(џџџџDocumentSummaryInformation8џџџџџџџџџџџџ$CompObjџџџџџџџџџџџџjџџџџџџџџџџџџџџџџџџџџџџџџўџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџџўџ џџџџ РFMicrosoft Word Document MSWordDocWord.Document.8є9Вq