Identification and expression analysis of novel LAGE-1 alleles with single nucleotide polymorphisms in cancer patients.

OBJECTIVE: The purpose of this study is to identify single nucleotide polymorphisms (SNPs) in the coding region alleles of the X chromosomal LAGE-1 gene, and investigate the frequency of such SNPs in both cancer patients and healthy controls, and thus determine the potential significance of these SNPs with respect to cancer vaccine therapy. METHODS: In this study, different mRNAs transcribed from the LAGE-1 gene were identified by RT-PCR from healthy donors and cancer patients samples. RESULTS: A new LAGE-1 allele containing three coding region SNPs (69A/G, 317C/G, and 397T/G) were identified. The allele is highly expressed as the LAGE-1a mRNA variant AY679089 in some of the cancer patients. The three SNPs altered the LAGE-1 gene sequence to that of NYESO-1 at both the nucleotide and amino acid level. CONCLUSION: There is a high frequency of the LAGE-1 gene allele with SNPs in coding regions in cancer patients. There was a clear relationship between the variant AY679089 and gastric cancer. The SNPs may lead to accelerated progress of poorly differentiated gastric cancer. The SNPs found in these alleles may also alter the immunological characteristics of LAGE-1a and should be taken into account if this antigen is adopted as a cancer vaccine component.

[Identification and expression of non-coding RNAs NC28 and NC119 in human tumors].

OBJECTIVE: To explore and identify the non-coding RNAs related to tumors. METHODS: We used RT-PCR and Northern blot to analyze non-coding RNAs in tumor tissues and cell lines. RESULTS: Two predicted non-coding RNAs were confirmed to be expressed in cancer tissues and cell lines by RT-PCR and DNA sequencing. We detected the expression of two non-coding RNA transcripts by Northern blot. The length of NC28 was about 1800 nt, and that of NC119 was about 1200nt. CONCLUSIONS: NC28 and NC119 have a tumor-associated expression pattern. The non-coding RNAs may play a role in the development of tumors.

[The expression of 11 cancer/testis (CT) antigen genes in esophageal carcinoma].

OBJECTIVE: To investigate the expression status of 11 different cancer/testis (CT) antigen genes in esophageal carcinoma. METHODS: Esophageal carcinoma tissue and adjacent normal esophageal mucosa taken from 35 esophageal carcinoma patients were assayed for the expression of 11 different CT antigen genes by RT-PCR techniques. RESULTS: Of the 11 CT antigen genes analyzed, none of them was expressed in normal esophageal mucosa. MAGE-3 was found to be the most frequently expressed in esophageal carcinoma tissues (62.9%), followed, in the order of expression frequency, by MAGE4 (31.4%), LAGE-1 (28.6%), MAGE-1 (25.7%), CT10 (20.0%), NY-ESO-1 (20.0%), CT7 (5.7%) and SCP1 (2.9%). No expression of SSX-1, SSX-2 and SSX-4 was found. Among the 35 cases, 28 (80.0%) expressed at least one CT antigen gene, 21 (60.0%) expressed more than 2 CT antigen genes, and 4 of the 21 (19.0%) expressed more than 4 CT antigens, which accounted for 11.4% of total number of patients (4/35). No CT antigen expression was found in the tumor tissue in 7 cases, including 5 cases in stage II and 1 case each in stage I and IV, respectively. Of the 11 CT genes examined, expression of 5 genes (NY-ESO-1, LAGE-1, MAGE-1, MAGE-3 and MAGE-4) was correlated with tumor progression. SCP-1 and CT10 expression was found more frequently in early stage patients. With progression of the disease, the frequency of co-expression of multiple CT antigen genes was significantly increased reaching 28.6% in stage III patients. CONCLUSION: Of the 11 different CT antigen genes examined by RT-PCR in esophageal carcinoma, 8 genes were detected in various frequencies in 28 of the 35 esophageal cancer patients studied. They are candidate tumor-associated antigens in the preparation of tumor vaccines for immunotherapy in esophageal cancer patients.

Identification of suitable reference genes in buffalo grass for accurate transcript normalization under various abiotic stress conditions.

Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) is a sensitive technique for normalization of the gene expression level of target genes. Buffalograss (Buchloe dactyloides), a warm-season turfgrass with strong abiotic stress resistance, is widely used in North China. Up to now, no work was performed to evaluate the reference genes in buffalograss. In this study, the expression profiles of ten potential reference genes were examined by qRT-PCR in 24 buffalograss samples, which were subjected to a different treatment (salt, osmotic, cold and heat). Three qRT-PCR analysis methods (GeNorm, NormFinder, and Bestkeeper) were used to evaluate the stability of gene expression. The results indicated that DNAJ and ß-ACTIN were the optimal reference genes for salt-treated leaves, and the combination of PP2A and GAPDH was better reference genes for PEG-treated leaves. Under cold stress, DNAJ and ß-ACTIN showed less variety of expression level in leaves. DNAJ and GAPDH exhibited the most stable expression in heat-treated samples. To sum up, glyceral-dehyde-3-phosphate dehydrogenase (GAPDH), ß-ACTIN, DNAJ-like protein (DNAJ) and protein phosphatase 2A (PP2A) were selected as the most stable reference gene among all tested samples. To further validate the suitability of these reference genes, the expression levels of DREB2 (homologs of AtDREB2) were analyzed in parallel. Our results show that the best reference genes differed across different experimental conditions, and these results should enable better normalization and quantification of transcript levels in buffalograss in the future.

[Population structure and spatial distribution pattern of Camellia azalea in E' huangzhang Nature Reserve of Guangdong, China].

This paper studied the structures of basal diameter, height and canopy width of Camellia azalea population in E' huangzhang Nature Reserve of Guangdong. The spatial distribution patterns and dynamics of the population were measured by applying aggregate indices including disperse coefficient, negative binomial distribution, Cassie index, clumping index, mean crowding, patch index and Green index. The results showed that in the natural distribution region, the population was mainly composed of adult trees, and was in declining due to seriously lack of seedlings. The structures of diameter, height and canopy width were not identical among different plots. In the plots, the spatial distribution pattern of C. azalea population showed clump or random, and changed from clump to random with the development of the population.

Phenotypic Responses of a Stoloniferous Clonal Plant Buchloe dactyloides to Scale-Dependent Nutrient Heterogeneity.

Clonal plants could modify phenotypic responses to nutrients heterogeneously distributed both in space and time by physiological integration. It will take times to do phenotypic responses to modifications which are various in different growth periods. An optimal phenotype is reached when there is a match between nutrient conditions and foraging ability. A single plantlet of Buchloe dactyloides with two stolons was transplanted into heterogeneous nutrient conditions. One stolon grew in homogeneous nutrient patch, while the other cultured in different scales of heterogeneous nutrient patches. As compared to the other nutrient treatment, heterogeneous nutrient treatments with small scale of 25×25 cm resulted in a higher biomass, and larger number of ramets, clumps and stolons in B. dactyloides at both genet and clonal fragment levels. Significant differences of number of ramets, clumps and stolons were detected at the rapid growth stage, but not in the early stage of the experiment. Foraging ability was more efficient in heterogeneous than in homogeneous nutrient conditions as assessed by higher root mass and root to shoot ratio. Different nutrient treatments did not prompt significant differences in internode and root length. Physiological integration significantly increased biomass, but did not influence other growth or morphological characters. These results suggest that physiological integration modifies phenotypic plasticity of B. dactyloides for efficient foraging of nutrients in heterogeneous nutrient conditions. These effects are more pronounced at genet and clonal fragment levels when the patch scale is 25×25 cm. Time is a key factor when phenotypic plasticity of B. dactyloides in heterogeneous nutrient conditions is examined.