A modified mutagenic PCR-RFLP method for K-ras codon 12 and 13 mutations detection in NSCLC patients

Hatzaki, A.; Razi, Evangelia; Anagnostopoulou, Katherine K.; Iliadis, K.; Kodaxis, A.; Papaioannou, D.; Labropoulos, Stefanos; Vasilaki, M.; Kosmidis, P.; Saetta, Angelica A.; Mihalatos, Markos; Nasioulas, Georgios

doi:10.1006/mcpr.2001.0367

Cited by 32 publications

(25 citation statements)

References 13 publications

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“…Genomic DNA was isolated from normal lung tissue as described previously (24), and the mutation status of codon 12 and codon 13 of KRAS was determined using RFLP analysis and sequencing as previously described (25,26).…”

Section: Methodsmentioning

confidence: 99%

Gene Expression Signature Predicts Recurrence in Lung Adenocarcinoma

Larsen

Pavey

Passmore

et al. 2007

Clinical Cancer Research

103

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Purpose: Improving outcomes for early-stage lung cancer is a major research focus at present because a significant proportion of stage I patients develop recurrent disease within 5 years of curative-intent lung resection. Within tumor stage groups, conventional prognostic indicators currently fail to predict relapse accurately. Experimental Design: To identify a gene signature predictive of recurrence in primary lung adenocarcinoma, we analyzed gene expression profiles in a training set of 48 node-negative tumors (stage I-II), comparing tumors from cases who remained disease-free for a minimum of 36 months with those from cases whose disease recurred within 18 months of complete resection. Results: Cox proportional hazards modeling with leave-one-out cross-validation identified a 54-gene signature capable of predicting risk of recurrence in two independent validation cohorts of 55 adenocarcinomas [log-rank P = 0.039; hazard ratio (HR), 2.2; 95% confidence interval (95% CI), 1.1-4.7] and 40 adenocarcinomas (log-rank P = 0.044; HR, 3.3; 95% CI, 1.4-7.9). Kaplan-Meier log-rank analysis found that predicted poor-outcome groups had significantly shorter survival, and furthermore, the signature predicted outcome independently of conventional indicators of tumor stage and node stage. In a subset of earliest stage adenocarcinomas, generally expected to have good outcome, the signature predicted samples with significantly poorer survival. Conclusions: We describe a 54-gene signature that predicts the risk of recurrent disease independently of tumor stage and which therefore has potential to refine clinical prognosis for patients undergoing resection for primary adenocarcinoma of the lung.

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Section: Methodsmentioning

confidence: 99%

Gene Expression Signature Predicts Recurrence in Lung Adenocarcinoma

Larsen

Pavey

Passmore

et al. 2007

Clinical Cancer Research

103

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“…We used a modified mutagenic PCR -RFLP method (Hatzaki et al, 2001) for screening mutations in codon 12 of the K-ras gene. PCR primers and the amplification conditions are shown in Table 2.…”

Section: Mutation Detection and Nucleotide Sequence Analysis Of The Ementioning

confidence: 99%

Mutations in the epidermal growth factor receptor gene are linked to smoking-independent, lung adenocarcinoma

et al. 2005

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Epidermal growth factor receptor (EGFR) mutations are a potential predictor of the effectiveness of EGFR inhibitors for the treatment of lung cancer. Although EGFR mutations were reported to occur with high frequency in nonsmoking Japanese adenocarcinoma patients, the exact nature has not been fully elucidated. We examined EGFR gene mutations within exons 18 -21 and their correlations to clinico-pathological factors and other genetic alterations in tumour specimens from 154 patients who underwent resection for lung cancer at Kyoto University Hospital. Epidermal growth factor receptor mutations were observed in 60 tumours (39.0%), all of which were adenocarcinoma. Among the patients with adenocarcinoma (n ¼ 108), EGFR mutations were more frequently observed in nonsmokers than former smokers or current smokers (83.0, 50.0, 15.2%, respectively), in women than men (76.3 vs 34.0%), in tumours with bronchio-alveolar component than those without bronchio-alveolar component (78.9 vs 42.9%), and in well or moderately differentiated tumours than poorly differentiated tumours (72.0, 64.4, 34.2%). No tumours with EGFR mutations had any K-ras codon 12 mutations, which were well-known smoking-related gene mutations. In conclusion, adenocarcinomas with EGFR mutation had a distinctive clinico-pathological feature unrelated to smoking. Epidermal growth factor receptor mutations may play a key role in the development of smoking-independent adenocarcinoma.

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“…Samples were dissolved in 0.8 mM NaOH. DNA was amplified by mutagenic PCR assay (24). A mismatched upstream primer for codon 12 and downstream primer for codon 13 were used to introduce the MvaI and BsuRI restriction sites, respectively.…”

Section: Methodsmentioning

confidence: 99%

Deregulated expression of the per2 gene in human colorectal carcinoma

Zeman

Vician²,

Monošíková³

et al. 2008

Mol Med Report

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Abstract. The circadian system is involved in the control of cell proliferation and apoptosis. The aim of this study was to analyze expression of the human per2 gene in patients who underwent surgery for colorectal carcinoma. The study included 25 patients of both genders. Patients were exposed to light from 6:00 a.m. until 9:00 p.m. according to standard hospital practice. Tissue samples were taken from the tumor as well as from the proximal and distal areas of the resected colon at the time of surgery. Surgery was performed during the morning hours. Expression of per2 mRNA was measured by real-time PCR. There was a significant negative correlation between per2 gene expression and tumor staging. Expression of per2 mRNA did not correlate with whether the tumor was localized in the colon or rectum. In comparison with ectomized tissue without malignancy from patients with colorectal carcinoma, our data demonstrate per2 mRNA deregulation in tumor tissue, and suggest a way in which the circadian system can influence tumorigenesis.

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A modified mutagenic PCR-RFLP method for K-ras codon 12 and 13 mutations detection in NSCLC patients

Cited by 32 publications

References 13 publications

Gene Expression Signature Predicts Recurrence in Lung Adenocarcinoma

Gene Expression Signature Predicts Recurrence in Lung Adenocarcinoma

Mutations in the epidermal growth factor receptor gene are linked to smoking-independent, lung adenocarcinoma

Deregulated expression of the per2 gene in human colorectal carcinoma

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