Epidermal Growth Factor Receptor Mutations and Their Prognostic Value with Carcinoembryonic Antigen in Pathological T1 Lung Adenocarcinoma.

AIMS: The prognostic value of epidermal growth factor receptor (EGFR) mutations in the context of serum carcinoembryonic antigen levels remains controversial in T1 lung adenocarcinoma. METHODS: Clinical and pathological characteristics, preoperational carcinoembryonic antigen levels, EGFR mutations, and disease-free and overall survival were analysed retrospectively in 573 pathological T1 patients in East China. RESULTS: EGFR mutations were detected in 220 of 573 patients (38.4%). Patients with serum carcinoembryonic antigen levels ≥ 2.12 ng/mL had worse disease-free (P < 0.001) and overall survival (P < 0.001) than had others, although survival was comparable between patients with and without EGFR mutations. However, patients with exon 21 mutations in EGFR had significantly better overall survival than had patients with exon 19 mutations (P = 0.016), although disease-free survival was comparable (P = 0.424). Among patients with serum carcinoembryonic antigen levels ≥ 2.12 ng/mL, disease-free (P = 0.019) and overall survival (P < 0.001) was also better than that in those with exon 21 mutations. Finally, the exon 19 deletion was found to be an independent predictor of unfavourable overall survival (P = 0.037). CONCLUSIONS: EGFR mutations were associated with preoperational serum carcinoembryonic antigen levels ≥ 2.12 ng/mL. In patients with levels above this threshold, those with the exon 19 deletion have less favourable prognosis than have those with the exon 21 mutation.

Characterization of a novel antimicrobial peptide with chitin-biding domain from Mytilus coruscus.

Using reverse phase high performance liquid chromatography (RP-HPLC), a novel antimicrobial peptide with 55 amino acid residues was isolated from the hemolymph of Mytilus coruscus. This new antimicrobial peptide displays predominant antimicrobial activity against fungi and Gram-positive bacteria. The molecular mass and the N-terminal sequence of this peptide were analyzed by Mass Spectrometry and Edman degradation, respectively. This antimicrobial peptide, with molecular mass of 6621.55 Da, is characterized by a chitin-biding domain and by 6 Cysteine residues engaged in three intra-molecular disulfide bridges. The full-length of cDNA sequence of this new peptide was obtained by rapid amplification of cDNA ends (RACE) and the encoded precursor was turn out to be a chitotriosidase-like protein. Therefore, we named the precursor with mytichitin-1 and the new antimicrobial peptide (designated as mytichitin-CB) is the carboxyl-terminal part of mytichitin-1. The mRNA transcripts of mytichitin-1 are mainly detected in gonad and the expression level of mytichitin-1 in gonad was up-regulated and reached the highest level at 12 h after bacterial challenge, which was 9-fold increase compared to that of the control group. These results indicated that mytichitin-1 was involved in the host immune response against bacterial infection and might contribute to the clearance of invading bacteria.

High expression of serum miR-21 and tumor miR-200c associated with poor prognosis in patients with lung cancer.

Serum microRNAs have been identified as potential cancer biomarkers. However, the detailed mechanism by which expression of microRNAs contributes to the development and diagnosis of NSCLC remains unknown. This study was to identify specific miRNAs for diagnosing or predicting the prognosis of NSCLC patients and their correlation between miRNA expression in tissues and serums. Six matched cancer and noncancerous tissues from NSCLC patients were analyzed by miRNA microarray. Among these, three miRNAs (miR-21, miR-141, and miR-200c) were examined in 70 NSCLC paired samples (cancer, normal tissue, and serum) and 44 serum samples of normal volunteers by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Consisting with the microarray results, the expression levels of miR-21, miR-141, and miR-200c in NSCLC were higher than those in normal tissues. While the level of serum miR-21 was increased in cancer patients as compared with that in normal counterpart, expression of miR-141 and miR-200c showed lower levels in serums from cancer patients. Overexpression of serum miR-21 was strongly associated with lymph node metastasis and advanced clinical stage of NSCLC. Finally, log-rank and Cox regression tests demonstrated that high expressions of tumor miR 21 and miR-200c or serum miR-21 were associated with a poor survival in NSCLC patients. Our results suggest that tumor miR-21, miR-141, miR-200c, and serum miR-21 may be potential novel biomarkers for the diagnosis of NSCLC. In addition, this study, for the first time, identifies a significant role of the tumor miR-200c played in predicting prognosis in patients with NSCLC.

Nicotine inhibits cisplatin-induced apoptosis in NCI-H446 cells.

Nicotine is not only a major component in tobacco but is also a survival agonist that inhibits apoptosis induced by certain agents including chemotherapeutic drugs. Here, we first showed that nicotine inhibits cisplatin-induced apoptosis in NCI-H446 cells. An MTT assay, Annexin V-FITC staining, RT-PCR, and Western blot were applied to identify the viability of cells, stages of apoptosis, mRNA and signaling proteins expression, respectively. First, we observed that nicotine induced no significant apoptosis when used alone and promoted cell proliferation at a low concentration or for a short time, but the opposite was observed at a high concentration or for a long time. In addition, an increase in XIAP and Survivin mRNA or protein was observed. Next, when combined with cisplatin, growth inhibition rates were concentration dependent, decreased to the lowest level at first, but later climbed to the highest point. Furthermore, nicotine inhibited apoptosis induced by cisplatin and caused a concentration-dependent increase in both XIAP and Survivin mRNA or protein. Moreover, the apoptotic effect of the combination group was obviously higher than that of nicotine used alone at the same nicotine concentration and lower than that of cisplatin used alone at the same cisplatin concentration. These studies suggest that exposure to nicotine might negatively impact the apoptotic potential of chemotherapeutics.

[Promoter methylation status of hPer3 gene in AML patients and the in vitro effect of decitabine on the status].

OBJECTIVE: To investigate the clinical significance of promoter methylation status of hPer3 gene in acute myeloid leukemia (AML) patients and the in vitro effect of decitabine (DCA) on AML cell lines HL-60 and U937. METHODS: The promoter methylation status of hPer3 gene and mRNA expression levels in bone marrow of 206 AML and 40 iron deficiency anemia (IDA) patients (as control) were detected by methylation specific PCR (MS-PCR) and real-time PCR (RT-PCR). The HL-60 and U937 cell lines were treated with different concentrations of DCA for 48 and 72 h. The inhibition rates of cell proliferation were detected by methyl thiazolyl tetrazolium (MTT); the early apoptosis rates by staining with Annexin V and PI; the CD14 and CD11b expressions by flow cytometry (FCM); the promoter methylation status of hPer3 gene by MS-PCR; and the hPer3 mRNA expressions levels by RT-PCR. RESULTS: The promoter methylation rates of hPer3 in newly diagnosed (ND) group, partial remission(PR) group, complete remission (CR) group, relapse (R) group and control group were 93.65% (59/63), 54.39% (31/57), 24.66% (18/73), 61.54% (8/13) and 0% (0/40), and the hPer3 mRNA expression levels were 0.19 ± 0.08, 6.28 ± 2.11, 52.76 ± 14.17, 8.18 ± 4.36, 75.03 ± 18.16, respectively. There was a significant statistic difference between any two group (P < 0.01) excepting for between PR and R group (P > 0.05). After DCA treatment, the promoter hypermethylation status of hPer3 was reduced and the mRNA and CD14, CD11b expression levels were up regulated in a dose dependent manner with an induction of cell apoptosis. CONCLUSIONS: Promotor methylation status and mRNA expression of hPer3 gene may be indicators for evaluating AML. DCA can induce the expression of hPer3 gene and cells apoptosis in AML.

[Expression and prognostic value of vacuole membrane protein 1 in non-small-cell lung cancer].

OBJECTIVE: To investigate the expression and prognostic value of vacuole membrane protein 1 (VMP1) in non-small-cell lung cancer (NSCLC). METHODS: The clinical data and survival status of 78 NSCLC patients were collected. Their paraffin-embedded tissues were detected immunohistochemically with a custom-made rabbit anti-human VMP1 polyclonal antibody. And the clinical significance of VMP1 expression and its relationship with the overall survival rate were analyzed statistically. RESULTS: The positive expression of VMP1 could only be observed in cytoplasm of cancer cells. Among 78 patients, 40 (51.3%) patients were stained VMP1-positive. The positive rate of VMP1 had a correlation with clinical stage (χ(2) = 6.829, P < 0.05), but not with gender, age, pathological type, gross classification and differentiation grade (P > 0.05). The overall 1-and 3-year survival rate was 94.9% and 66.3% respectively. The overall estimated survival time was 37.2 months (95%CI: 33.7 - 40.8). The expression of VMP1 had significant influence on the survival rate (χ(2) = 6.192, P < 0.05). The VMP1-positive patients lived shorter with an estimated survival time of 29.0 months (95%CI: 25.0 - 33.0). VMP1 expression and clinical stage were independent risk factors of influencing the survival rate. CONCLUSION: The detection of VMP1 in resected NSCLC tumor tissues may be helpful for prognostic prediction. NSCLC patients with VMP1-positive or late clinical stage have a worse prognosis.

[Sequence polymorphism of the promoter region of gene STK11 in patients with Peutz-Jeghers syndrome].

OBJECTIVE: To explore the relationship between the sequence variation of the promoter region (-1543 approximately -1160) of STK11 gene and the risk of developing Peutz-Jeghers syndrome (PJS). METHODS: The sequences of the promoter region of 14 PJS patients (7 patients are inherited and the other 7 patients are sporadic) and 42 normal individuals were PCR amplified and then sequenced. RESULTS: A new single nucleotide polymorphism (SNP) G/T (-1275) in STK11 promoter region was identified. The frequency of genotype GG, GT, and TT was 53.3%, 26.7%, and 20%, respectively among PJS patients and 33.3%, 64.3%, and 2.4%, respectively among the normal individuals. The frequency of genotype GG and TT among patients was significantly higher than that among the normal individuals, and the frequency of genotype GT among patients was significantly lower than that among the normal individuals (chi(2)=8.521, P<0.05). CONCLUSION: G/T(-1275) in STK11 promoter region is a new SNP. The genotype of this new SNP may relate to the risk of developing Peutz-Jeghers syndrome (PJS) deserve further research.