Expression and significance of IGF-1 and IGF-1R in thyroid nodules.

There are limitations in the diagnosis of thyroid nodules, especially follicular lesions, and their pathogenesis remains unclear. Insulin-like growth factor-1 (IGF-1) has been implicated in tumor cell apoptosis, transformation, invasion, and metastasis; however, its role in thyroid nodules is undetermined. The aim of this study is to investigate the relationship between expression of IGF-1 and thyroid nodule, and evaluate the role of IGF-1 in differential diagnosis and pathogenetic function of benign and malignant thyroid nodules. Sixty-two paraffin-embedded thyroid tissues from patients with thyroid nodules were selected, including 18 follicular adenomas (FA), 17 nodular goiters, 13 papillary thyroid carcinomas (PTC), 2 follicular thyroid carcinomas, and 12 normal controls. IGF-1 and IGF-1R protein and mRNA expression was detected by immunohistochemistry (IHC) and real-time quantitative PCR (qRT-PCR), respectively. Grouping comparisons were employed among the above groups based on the clinical and pathological subtypes. IGF-1 and IGF-1R expression was significantly higher in FA, nodular goiters, and PTC than that in the controls (all P < 0.01). Similarly, IGF-1 mRNA expression was also significantly higher in FA (P < 0.05), nodular goiters (P < 0.01), and PTC (P < 0.01) compared with the controls. IGF-1R mRNA expression was also significantly higher in FA, nodular goiters, and PTC (all P < 0.01) compared with the controls. Compared with FA and nodular goiters, PTC showed significantly higher IGF-1 and IGF-1R protein (P < 0.01, P < 0.05, respectively) and mRNA expression (P < 0.01, P < 0.05, respectively). IGF-1 probably plays an important role in the genesis and development of certain solid cold thyroid nodules, including PTC, nodular goiters, and FA. Detection of IGF-1 and IGF-1R expression in thyroid tissues by IHC or qRT-PCR is hard to distinguish malignant from benign lesions.

[A new method for 5, 10-methylenetetrahydrofolate reductase single nucleotide polymorphisms genotyping used to study susceptibility of hematological malignancy].

The aim of this study was to set up a new method for 5, 10-Methylenetetrahydrofolate reductase (MTHFR) single nucleotide polymorphisms (SNP) genotyping, and to investigate the hereditary susceptibility of hematological malignancy. Prepared an aimed gene microarray based on cDNA microarray theory, dual-color fluorescence hybridization was used to detect SNP loci, and DNA sequencing was performed to confirm the results. The MTHFR C677T SNP loci of 157 controls and 127 patients with hematological malignancies (30 multiple myeloma, 28 non-Hodgkin's lymphoma, 22 acute lymphoblastic leukemia, 40 acute myeloid leukemia, 7 chronic myeloid leukemia) from Jiangsu province were detected. The results showed that after overlapping, homozygous wild type, heterozygote type and homozygous mutant type yielded green, yellow and red fluorescence, respectively. DNA sequencing validated these results. The allele frequency of 677C and 677T in patients and controls were 58.7% and 66.9%, 41.3% and 33.1% respectively, showing statistically significant difference (chi2 = 4.077, P = 0.043). 677TT genotype showed a significantly higher risk of MM (OR = 4.21; 95% CI = 1.50 - 11.83; P = 0.006). It is concluded that this microarray-based method is accurate, high-throughput and inexpensive, suitable for SNP genotyping in a large number of individuals. C677T polymorphisms influence the risk of hematological malignancies. 677TT genotype is susceptive to MM.

Detection of p16 hypermethylation in gastric carcinomas using a seminested methylation-specific PCR.

Aberrant DNA methylation of a CpG site is among the earliest and most frequent alterations in various tumors including gastric carcinoma. The aim of this study is to detect tumor-associated aberrant hypermethylation of the p16 gene from 60 gastric tumor and corresponding normal tissues using a seminested methylation-specific PCR (MSP). The results indicated that hypermethylation of the p16 gene could be detected in 80% (48/60) of the gastric tumor samples from the first PCR. However, the frequency increased significantly to 86.7% (52/60) of the gastric tumor samples after the second PCR. These results show that this technique increases the sensitivity of detecting p16 hypermethylation from tumor samples. Furthermore, the aberrant methylation of p16 was observed in all of the stages, confirming that this epigenetic alteration is an early event during gastric carcinogenesis. Clinicopathologic parameters such as age, sex, and histological differentiation of GC were not significantly associated with the methylation status.

Microarray-based method for detecting methylation changes of p16(Ink4a) gene 5'-CpG islands in gastric carcinomas.

AIM: Aberrant DNA methylation of CpG site is among the earliest and most frequent alterations in cancer. Several studies suggest that aberrant methylation of the CpG sites of the tumor suppressor gene is closely associated with carcinogenesis. However, large-scale analysis of candidate genes has so far been hampered by the lack of high-throughput approach for analyzing DNA methylation. The aim of this study was to describe a microarray-based method for detecting changes of DNA methylation in cancer. METHODS: This method used bisulfite-modified DNA as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. Therefore, the amplified product might contain a pool of DNA fragments with altered nucleotide sequences due to differential methylation status. Nine sets of oligonucleotide probes were designed to fabricate a DNA microarray to detect the methylation changes of p16 gene CpG islands in gastric carcinomas. The results were further validated by methylation-specific PCR (MSP). RESULTS: The experimental results showed that the microarray assay could successfully detect methylation changes of p16 gene in 18 gastric tumor samples. Moreover, it could also potentially increase the frequency of detecting p16 methylation from tumor samples than MSP. CONCLUSION: Microarray assay could be applied as a useful tool for mapping methylation changes in multiple CpG loci and for generating epigenetic profiles in cancer.