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Article: Aberrant DNA methylation in non-neoplastic gastric mucosa of H. Pylori infected patients and effect of eradication.

Perri, Francesco / Cotugno, Rosa / Piepoli, Ada / Merla, Antonio / Quitadamo, Michele / Gentile, Annamaria / Pilotto, Alberto / Annese, Vito / Andriulli, Angelo

The American journal of gastroenterology

2007 Volume 102, Issue 7, Page(s) 1361–1371

Abstract: Background: Gene promoter methylation is an epigenetic event leading to gene silencing. This mechanism is particularly relevant in cancer since it can interfere with the activity of specific "suppressor" genes.: Aim: To evaluate promoter methylation ... ...

Abstract	Background: Gene promoter methylation is an epigenetic event leading to gene silencing. This mechanism is particularly relevant in cancer since it can interfere with the activity of specific "suppressor" genes. Aim: To evaluate promoter methylation of CDH1, p16, APC, MLH1, and COX2 in patients with H. pylori (Hp) infection before and after eradication. Methods: Fifty-seven dyspeptic outpatients who had never performed previous endoscopy or Hp testing and treatment underwent clinical interview, endoscopy with three paired gastric biopsy specimens from the antrum, angulus, and corpus, and (13)C-urea breath test (UBT). Biopsies were scored for the presence of Hp and intestinal metaplasia (IM). DNA methylation of five tumor-related genes (CDH1, p16, MLH1, APC, and COX2) was evaluated by methylation-specific PCR in each biopsy. Infected patients were given a standard eradicating treatment and, after 1 yr, underwent endoscopy with biopsies and UBT. Results: Hp infection was found in 45 patients. IM was detected in 17 out of 45 (38%) infected patients. Mean number of methylated genes was 0, 1.1 +/- 0.9, and 1.6 +/- 0.9 among the 12 Hp-/IM-, the 28 Hp+/IM-, and the 17 Hp+/IM+ patients, respectively (P < 0.0001). Specifically, promoter hypermethylation of CDH1, p16, APC, MLH1, and COX2 was found in 68%, 25%, 7%, 0%, and 14% of Hp+/IM- patients and in 71%, 29%, 35%, 12%, and 12% of Hp+/IM+ patients. No significant difference was found among the three groups of patients as far as age, smoking, alcohol, meat and vegetable consumption, and family history of gastric cancer were considered. Twenty-three out of 45 (51%) infected patients underwent the 1-yr follow-up endoscopy: 17 out of 23 (74%) were successfully eradicated. After Hp eradication, CDH1, p16, and APC methylation significantly decreased while COX2 methylation completely disappeared. Conversely, MLH1 methylation did not change significantly in patients with IM. Conclusion: Hp infection is associated with promoter methylation of genes which are relevant in the initiation and progression of gastric carcinogenesis. While CDH1 methylation seems to be an early event in Hp gastritis, MLH1 methylation occurs late along with IM. Hp eradication is able to significantly reduce gene methylation thus delaying or reversing Hp-induced gastric carcinogenesis.
MeSH term(s)	Adaptor Proteins, Signal Transducing/genetics ; Adult ; Aged ; Anti-Bacterial Agents/therapeutic use ; Biopsy ; Cadherins/genetics ; Cyclooxygenase 2/genetics ; DNA/genetics ; DNA Methylation ; DNA Repair ; Endoscopy, Gastrointestinal ; Female ; Follow-Up Studies ; Gastric Mucosa/metabolism ; Gastric Mucosa/microbiology ; Gastric Mucosa/pathology ; Gastritis/drug therapy ; Gastritis/genetics ; Gastritis/microbiology ; Gene Silencing ; Genes, APC ; Genes, Tumor Suppressor ; Genes, p16 ; Helicobacter Infections/drug therapy ; Helicobacter Infections/genetics ; Helicobacter Infections/microbiology ; Helicobacter pylori/isolation & purification ; Humans ; Male ; Membrane Proteins/genetics ; Middle Aged ; MutL Protein Homolog 1 ; Nuclear Proteins/genetics ; Polymerase Chain Reaction
Chemical Substances	Adaptor Proteins, Signal Transducing ; Anti-Bacterial Agents ; CDH1 protein, human ; Cadherins ; MLH1 protein, human ; Membrane Proteins ; Nuclear Proteins ; DNA (9007-49-2) ; Cyclooxygenase 2 (EC 1.14.99.1) ; PTGS2 protein, human (EC 1.14.99.1) ; MutL Protein Homolog 1 (EC 3.6.1.3)
Language	English
Publishing date	2007-07
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	390122-1
ISSN	1572-0241 ; 0002-9270
ISSN (online)	1572-0241
ISSN	0002-9270
DOI	10.1111/j.1572-0241.2007.01284.x
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Article ; Online: Promoter methylation correlates with reduced NDRG2 expression in advanced colon tumour

D'Addabbo Annarita / Maglietta Rosalia / Carella Massimo / Panza Anna / Merla Antonio / Quitadamo Michele / Augello Bartolomeo / Gentile Annamaria / Merla Giuseppe / Cotugno Rosa / Piepoli Ada / Ancona Nicola / Fusilli Saverio / Perri Francesco / Andriulli Angelo

BMC Medical Genomics, Vol 2, Iss 1, p

2009 Volume 11

Abstract: Abstract Background Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism ... ...

Abstract	Abstract Background Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism usually leads to inactivation of tumour-suppressor genes. We have designed the current study to validate our previous microarray data and to identify novel hypermethylated gene promoters. Methods The validation assay was performed in a different set of 8 patients with colorectal cancer (CRC) by means quantitative reverse-transcriptase polymerase chain reaction analysis. The differential RNA expression profiles of three CRC cell lines before and after 5-aza-2'-deoxycytidine treatment were compared to identify the hypermethylated genes. The DNA methylation status of these genes was evaluated by means of bisulphite genomic sequencing and methylation-specific polymerase chain reaction (MSP) in the 3 cell lines and in tumour tissues from 30 patients with CRC. Results Data from our previous genome search have received confirmation in the new set of 8 patients with CRC. In this validation set six genes showed a high induction after drug treatment in at least two of three CRC cell lines. Among them, the N-myc downstream-regulated gene 2 ( NDRG2) promoter was found methylated in all CRC cell lines. NDRG2 hypermethylation was also detected in 8 out of 30 (27%) primary CRC tissues and was significantly associated with advanced AJCC stage IV. Normal colon tissues were not methylated. Conclusion The findings highlight the usefulness of combining gene expression patterns and epigenetic data to identify tumour biomarkers, and suggest that NDRG2 silencing might bear influence on tumour invasiveness, being associated with a more advanced stage.
Keywords	Internal medicine ; RC31-1245 ; Genetics ; QH426-470
Subject code	616
Language	English
Publishing date	2009-03-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
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Article: Amoxicillin/tetracycline combinations are inadequate as alternative therapies for Helicobacter pylori infection.

Perri, Francesco / Festa, Virginia / Merla, Antonio / Quitadamo, Michele / Clemente, Rocco / Andriulli, Angelo

Helicobacter

2002 Volume 7, Issue 2, Page(s) 99–104

Abstract: Background: Triple therapy with proton pump inhibitors or ranitidine bismuth citrate, clarithromycin and either amoxicillin or nitroimidazole derivatives are the present gold standards for cure of Helicobacter pylori infection. However, primary ... ...

Abstract	Background: Triple therapy with proton pump inhibitors or ranitidine bismuth citrate, clarithromycin and either amoxicillin or nitroimidazole derivatives are the present gold standards for cure of Helicobacter pylori infection. However, primary resistance to either clarithromycin or nitroimidazole derivatives is increasing and alternative therapies are needed. Aim: To determine the efficacy and safety of three regimens consisting of amoxicillin and tetracycline or doxycycline combined with either lansoprazole or ranitidine bismuth citrate. Methods: Two hundred and seventy H. pylori infected patients were randomly given one of the following treatments: amoxicillin 1 g twice a day (b.i.d.) plus tetracycline 500 mg four times a day (q.i.d.) with either lansoprazole 30 mg b.i.d. (group LAT) or ranitidine bismuth citrate 400 mg b.i.d. (group RBCAT) for 7 days and amoxicillin 1 g b.i.d. plus doxycycline 100 mg b.i.d. and lansoprazole 30 mg b.i.d. for 14 days (group LAD). Eradication rate was assessed by UBT at 4-6 weeks after therapy. Results: The three groups (LAT, RBCAT, and LAD) of patients achieved eradication rates of 35% (25-45), 20% (12-29) and 36% (25-46), respectively, on intention-to-treat analysis. Patient compliance was optimal and side-effects minimal in all three groups. Conclusions: Although the amoxicillin/tetracycline combination is attractive (inexpensive, safe, and with low primary resistance rate), it can not be recommended for H. pylori eradication.
MeSH term(s)	2-Pyridinylmethylsulfinylbenzimidazoles ; Adult ; Amoxicillin/therapeutic use ; Anti-Bacterial Agents/therapeutic use ; Anti-Ulcer Agents/therapeutic use ; Bismuth/therapeutic use ; Doxycycline/therapeutic use ; Drug Therapy, Combination ; Female ; Helicobacter Infections/drug therapy ; Helicobacter pylori ; Humans ; Lansoprazole ; Male ; Middle Aged ; Omeprazole/analogs & derivatives ; Omeprazole/therapeutic use ; Penicillins/therapeutic use ; Prospective Studies ; Ranitidine/analogs & derivatives ; Ranitidine/therapeutic use ; Tetracycline/therapeutic use ; Treatment Failure
Chemical Substances	2-Pyridinylmethylsulfinylbenzimidazoles ; Anti-Bacterial Agents ; Anti-Ulcer Agents ; Penicillins ; Lansoprazole (0K5C5T2QPG) ; ranitidine bismuth citrate (7AJ51I17KG) ; Amoxicillin (804826J2HU) ; Ranitidine (884KT10YB7) ; Tetracycline (F8VB5M810T) ; Omeprazole (KG60484QX9) ; Doxycycline (N12000U13O) ; Bismuth (U015TT5I8H)
Language	English
Publishing date	2002-04-19
Publishing country	England
Document type	Clinical Trial ; Journal Article ; Randomized Controlled Trial
ZDB-ID	1330665-0
ISSN	1523-5378 ; 1083-4389
ISSN (online)	1523-5378
ISSN	1083-4389
DOI	10.1046/j.1083-4389.2002.00066.x
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Article: Lack of association between UGT1A7, UGT1A9, ARP, SPINK1 and CFTR gene polymorphisms and pancreatic cancer in Italian patients.

Piepoli, Ada / Gentile, Annamaria / Valvano, Maria Rosa / Barana, Daniela / Oliani, Cristina / Cotugno, Rosa / Quitadamo, Michele / Andriulli, Angelo / Perri, Francesco

World journal of gastroenterology

2005 Volume 12, Issue 39, Page(s) 6343–6348

Abstract: Aim: To investigate simultaneously UGT1A7, UGT1A9, ARP, SPINK and CFTR genes to verify whether genetic polymorphisms predispose to the development of pancreatic cancer (PC).: Methods: Genomic DNA of 61 pancreatic cancer patients and 105 healthy ... ...

Abstract	Aim: To investigate simultaneously UGT1A7, UGT1A9, ARP, SPINK and CFTR genes to verify whether genetic polymorphisms predispose to the development of pancreatic cancer (PC). Methods: Genomic DNA of 61 pancreatic cancer patients and 105 healthy controls (HC) were analyzed. UGT1A7 genotyping was determined by PCR-RFLP analysis. Specific PCR and sequencing were used to analyze genetic variants of UGT1A9, ARP, SPINK1 and CFTR genes. Results: Four different alleles (1: WT; 2: N129K and R131K; 3: N129K, R131K, and W208R; and 4: W208R) in UGT1A7 and three different alleles (1: WT; 4: Y242X; and 5: D256N) in UGT1A9 were detected. All UGT1A polymorphisms were observed at similar frequency in PC patients and HC. Seven different alleles in ARP were found in PC patients and HC at similar frequency. The SPINK1 mutations N34S and P55S occurred in five PC patients with a prevalence (4.1%) not significantly different from that observed (2.0%) in HC. The only CFTR DeltaF508 mutation was recognized in three PC patients with a prevalence (4.9%) similar to HC. Conclusion:* UGT1A7, UGT1A9, ARP, SPINK1 and CFTR gene polymorphisms are not associated with PC in Italian patients.
MeSH term(s)	Adult ; Age Factors ; Aged ; Aged, 80 and over ; Carrier Proteins/genetics ; Case-Control Studies ; Cystic Fibrosis Transmembrane Conductance Regulator/genetics ; Female ; Genetic Predisposition to Disease/epidemiology ; Genetic Predisposition to Disease/genetics ; Glucuronosyltransferase/genetics ; Humans ; Italy/epidemiology ; Male ; Middle Aged ; Mutation/genetics ; Nerve Growth Factors ; Pancreatic Neoplasms/genetics ; Polymorphism, Genetic ; Proteins/genetics ; Risk Factors ; Smoking ; Trypsin Inhibitor, Kazal Pancreatic
Chemical Substances	CFTR protein, human ; Carrier Proteins ; MANF protein, human ; Nerve Growth Factors ; Proteins ; SPINK1 protein, human ; Cystic Fibrosis Transmembrane Conductance Regulator (126880-72-6) ; Trypsin Inhibitor, Kazal Pancreatic (50936-63-5) ; Glucuronosyltransferase (EC 2.4.1.17) ; UDP-glucuronosyltransferase 1A9 (EC 2.4.1.17) ; UDP-glucuronosyltransferase, UGT1A7 (EC 2.4.1.17)
Language	English
Publishing date	2005-07-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2185929-2
ISSN	2219-2840 ; 1007-9327
ISSN (online)	2219-2840
ISSN	1007-9327
DOI	10.3748/wjg.v12.i39.6343
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Article: Comparison of a monoclonal antigen stool test (Hp StAR) with the 13C-urea breath test in monitoring Helicobacter pylori eradication therapy.

Perri, Francesco / Quitadamo, Michele / Ricciardi, Rosalba / Piepoli, Ada / Cotugno, Rosa / Gentile, Annamaria / Pilotto, Alberto / Andriulli, Angelo

World journal of gastroenterology

2005 Volume 11, Issue 37, Page(s) 5878–5881

Abstract: Aim: To evaluate the agreement between a mAb-based stool test (HP StAR) and the urea breath test (UBT) in monitoring (H pylori) infection after eradication therapy.: Methods: Patients with discordant results on UBT and Hp StAR underwent endoscopy ... ...

Abstract	Aim: To evaluate the agreement between a mAb-based stool test (HP StAR) and the urea breath test (UBT) in monitoring (H pylori) infection after eradication therapy. Methods: Patients with discordant results on UBT and Hp StAR underwent endoscopy with biopsies for rapid urease test, culture, and histology to confirm H pylori status. Results: Among 250 patients (50+/-14 years), 240 (96.0%) had concordant UBT and Hp StAR tests with a significant correlation between DOB and A values (R = 0.87; P<0.0001). The remaining 10 (4.0%) patients had discordant tests (positive Hp StAR and negative UBT) with the Hp StAR inaccurate in five cases (false positive) and UBT inaccurate in the other five cases (false negative). The "maximal expected" sensitivity, specificity, +PV, -PV, +LR, and -LR were 91%, 100%, 100%, 97.4%, infinity, and 8.2 respectively, for the UBT, and 100%, 97.4%, 91%, 100%, 38.8, and 0, respectively, for the Hp StAR. Overall accuracy for both tests was 98%. Conclusion: Both the UBT and the Hp StAR are equally accurate in monitoring H pylori infection. Nowadays, the choice of the "best" non-invasive H pylori test in the post-treatment setting should be done not only in terms of diagnostic accuracy but also in view of cost and local facilities.
MeSH term(s)	Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Monoclonal/immunology ; Antigens, Bacterial/immunology ; Breath Tests ; Feces/chemistry ; Helicobacter Infections/diagnosis ; Helicobacter Infections/drug therapy ; Helicobacter Infections/prevention & control ; Helicobacter pylori/immunology ; Humans ; Immunologic Tests/economics ; Immunologic Tests/methods ; Immunologic Tests/standards ; Male ; Middle Aged ; Monitoring, Physiologic ; Sensitivity and Specificity ; Urea/analysis ; Urease/metabolism
Chemical Substances	Antibodies, Monoclonal ; Antigens, Bacterial ; Urea (8W8T17847W) ; Urease (EC 3.5.1.5)
Language	English
Publishing date	2005-07-20
Publishing country	United States
Document type	Comparative Study ; Evaluation Studies ; Journal Article
ZDB-ID	2185929-2
ISSN	2219-2840 ; 1007-9327
ISSN (online)	2219-2840
ISSN	1007-9327
DOI	10.3748/wjg.v11.i37.5878
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Article ; Online: Promoter methylation correlates with reduced NDRG2 expression in advanced colon tumour.

Piepoli, Ada / Cotugno, Rosa / Merla, Giuseppe / Gentile, Annamaria / Augello, Bartolomeo / Quitadamo, Michele / Merla, Antonio / Panza, Anna / Carella, Massimo / Maglietta, Rosalia / D'Addabbo, Annarita / Ancona, Nicola / Fusilli, Saverio / Perri, Francesco / Andriulli, Angelo

BMC medical genomics

2009 Volume 2, Page(s) 11

Abstract: Background: Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism usually ... ...

Abstract	Background: Aberrant DNA methylation of CpG islands of cancer-related genes is among the earliest and most frequent alterations in cancerogenesis and might be of value for either diagnosing cancer or evaluating recurrent disease. This mechanism usually leads to inactivation of tumour-suppressor genes. We have designed the current study to validate our previous microarray data and to identify novel hypermethylated gene promoters. Methods: The validation assay was performed in a different set of 8 patients with colorectal cancer (CRC) by means quantitative reverse-transcriptase polymerase chain reaction analysis. The differential RNA expression profiles of three CRC cell lines before and after 5-aza-2'-deoxycytidine treatment were compared to identify the hypermethylated genes. The DNA methylation status of these genes was evaluated by means of bisulphite genomic sequencing and methylation-specific polymerase chain reaction (MSP) in the 3 cell lines and in tumour tissues from 30 patients with CRC. Results: Data from our previous genome search have received confirmation in the new set of 8 patients with CRC. In this validation set six genes showed a high induction after drug treatment in at least two of three CRC cell lines. Among them, the N-myc downstream-regulated gene 2 (NDRG2) promoter was found methylated in all CRC cell lines. NDRG2 hypermethylation was also detected in 8 out of 30 (27%) primary CRC tissues and was significantly associated with advanced AJCC stage IV. Normal colon tissues were not methylated. Conclusion: The findings highlight the usefulness of combining gene expression patterns and epigenetic data to identify tumour biomarkers, and suggest that NDRG2 silencing might bear influence on tumour invasiveness, being associated with a more advanced stage.
Language	English
Publishing date	2009-03-03
Publishing country	England
Document type	Journal Article
ISSN	1755-8794
ISSN (online)	1755-8794
DOI	10.1186/1755-8794-2-11
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Article ; Online: Antisecretory vs. antiproteasic drugs in the prevention of post-ERCP pancreatitis: the evidence-based medicine derived from a meta-analysis study.

Andriulli, Angelo / Caruso, Nazario / Quitadamo, Michele / Forlano, Rosario / Leandro, Gioacchino / Spirito, Fulvio / De Maio, Giovanni

JOP : Journal of the pancreas

2003 Volume 4, Issue 1, Page(s) 41–48

Abstract: Uncertainties still exist about the clinical benefit of pharmacological prevention of post-ERCP pancreatitis by either antisecretory drugs such as somatostatin and its long-acting analogue octreotide, or protease inhibitors such as gabexate mesilate. ... ...

Abstract	Uncertainties still exist about the clinical benefit of pharmacological prevention of post-ERCP pancreatitis by either antisecretory drugs such as somatostatin and its long-acting analogue octreotide, or protease inhibitors such as gabexate mesilate. Recent, large-scale prospective studies have reported a fourfold reduction in acute pancreatitis as compared to a placebo with the prophylactic administration of either gabexate mesilate or somatostatin, whereas octreotide was found to be ineffective. An initial meta-analysis of all available controlled trials on this topic has confirmed these findings. The indiscriminate use of these drugs in all patients is unlikely to be cost-effective, but the selective use of prophylaxis for high-risk patients might be advocated. Moreover, inasmuch as 85% of complications developed within 4 to 6 hours of completing the ERCP, it would be reasonable to infuse drugs only for this limited length of time. A recent prospective trial, carried out on high-risk patients, has surprisingly documented a higher incidence, although a non-significant one, of pancreatitis in patients who received short-term prophylaxis with somatostatin or gabexate mesilate than those given a placebo: 11.5% and 8.1% vs. 6.5%, respectively. In order to explore this discrepancy, the original meta-analysis was updated by including data of this negative trial: heterogeneity among the trials was apparent. A careful scrutiny of the most recent studies has revealed differences in patient population, protocols of drug administration, technique and operator-related risk factors for complications among the trials, which could explain, by themselves, the contrasting results reported by the interventional studies. In conclusion, current literature does not support the prophylactic use of either somatostatin or gabexate mesilate for the prevention of ERCP-related pancreatic damage, even in patients deemed to be at high risk for complications. At present, post-ERCP complications (and pancreatitis) can be prevented efficaciously by appropriate selection of patients, mastering of the technique and operator competence.
MeSH term(s)	Cholangiopancreatography, Endoscopic Retrograde/adverse effects ; Cholangiopancreatography, Endoscopic Retrograde/methods ; Clinical Competence ; Drug Administration Schedule ; Evidence-Based Medicine/methods ; Growth Hormone/antagonists & inhibitors ; Growth Hormone/secretion ; Humans ; Octreotide/administration & dosage ; Octreotide/therapeutic use ; Pancreatitis/diagnosis ; Pancreatitis/enzymology ; Pancreatitis/etiology ; Pancreatitis/prevention & control ; Patient Selection ; Protease Inhibitors/administration & dosage ; Protease Inhibitors/therapeutic use ; Randomized Controlled Trials as Topic ; Somatostatin/administration & dosage ; Somatostatin/analogs & derivatives ; Somatostatin/therapeutic use ; Treatment Outcome
Chemical Substances	Protease Inhibitors ; Somatostatin (51110-01-1) ; Growth Hormone (9002-72-6) ; Octreotide (RWM8CCW8GP)
Language	English
Publishing date	2003-01
Publishing country	Italy
Document type	Comparative Study ; Journal Article ; Meta-Analysis
ZDB-ID	2039637-5
ISSN	1590-8577 ; 1590-8577
ISSN (online)	1590-8577
ISSN	1590-8577
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Article: Genotyping of the lactase-phlorizin hydrolase c/t-13910 polymorphism by means of a new rapid denaturing high-performance liquid chromatography-based assay in healthy subjects and colorectal cancer patients.

Piepoli, Ada / Schirru, Enrico / Mastrorilli, Angela / Gentile, Annamaria / Cotugno, Rosa / Quitadamo, Michele / Merla, Antonio / Congia, Mauro / Usai Satta, Paolo / Perri, Francesco

Journal of biomolecular screening

2007 Volume 12, Issue 5, Page(s) 733–739

Abstract: Adult-type hypolactasia results from the progressive decline of lactase-phlorizin hydrolase activity in enterocytes after weaning. Lactase nonpersistence may determine a primary lactose intolerance with reduced diary product consumption, which is ... ...

Abstract	Adult-type hypolactasia results from the progressive decline of lactase-phlorizin hydrolase activity in enterocytes after weaning. Lactase nonpersistence may determine a primary lactose intolerance with reduced diary product consumption, which is possibly related to an increased risk of colon cancer. Recently, a genetic variant C/T(-13910) upstream of the lactase-phlorizin hydrolase (LCT) gene has been strongly correlated with the lactase persistence/nonpersistence trait in both family and case-control studies. The authors validate a denaturing high-performance liquid chromatography (dHPLC)-based assay versus conventional genotype sequencing in detecting the C/T(-13910) polymorphism of LCT and evaluate its prevalence in 2 different Italian geographical areas and in colorectal cancer patients. DNA samples of 157 healthy subjects and 124 colon cancer patients from Apulia and of 97 healthy subjects from Sardinia were evaluated for the C/T(-13910) polymorphism by dHPLC, sequencing, and restriction fragment length polymorphism (RFLP). Under optimized conditions, dHPLC was as sensitive as DNA sequencing and detected a new genetic variant (T/C(-13913)) in 2 individuals that was not identified by RFLP assay. Frequency of lactase nonpersistence genotype (C/C(-13910)) was similar in healthy subjects from 2 different Italian geographical areas and not increased in patients with colorectal cancer. The results indicate that the dHPLC method may be used as a rapid, noninvasive, and labor-saving screening tool for genotyping C/T(-13910) polymorphism, with high success, low cost, and reproducibility.
MeSH term(s)	Adult ; Biological Assay/methods ; Case-Control Studies ; Chromatography, High Pressure Liquid/methods ; Colorectal Neoplasms/enzymology ; Colorectal Neoplasms/genetics ; Colorectal Neoplasms/pathology ; Female ; Genetic Variation ; Genotype ; Heterozygote ; Homozygote ; Humans ; Italy/epidemiology ; Lactase/deficiency ; Lactase/genetics ; Lactase/metabolism ; Lactase-Phlorizin Hydrolase/genetics ; Lactose Intolerance/enzymology ; Lactose Intolerance/genetics ; Male ; Middle Aged ; Neoplasm Staging ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Prevalence ; Reproducibility of Results ; Sensitivity and Specificity ; Time Factors
Chemical Substances	Lactase (EC 3.2.1.108) ; Lactase-Phlorizin Hydrolase (EC 3.2.1.62)
Language	English
Publishing date	2007-08
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1433680-7
ISSN	1552-454X ; 1087-0571
ISSN (online)	1552-454X
ISSN	1087-0571
DOI	10.1177/1087057107301328
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