[Investigation of mRNA expression of collagen genes in oral squamous cell carcinoma and paired normal tissue].

OBJECTIVE: To investigate the mRNA expression of collagen genes in oral squamous cell carcinoma (OSCC) and paired normal oral mucosal tissue. METHODS: The differential mRNA expressions of collagen genes between 30 OSCC tissues and the paired normal oral mucosal tissues were detected by RT-PCR. RESULT: The relative expression level of COL1A1 mRNA in the 30 cancerous tissues was up-regulated by 2.78 folds as compared with its expression in the paired normal samples, suggesting its significant overexpression in OSCC (P<0.001). The expression levels of COL1A2, COL4A1, COL4A2, and COL5A2 mRNA in the cancerous tissues were up-regulated by 1.07, 1.15, 1.27, and 1.16 folds compared to those in paired normal samples (P>0.05). CONCLUSION: COL1A1 mRNA overexpression may play an important role in the carcinogenesis of OSCC and can be a potential molecular marker of OSCC.

[Pectoralis major myocutaneous flap for repairing large tissue defects following oral cancer surgery].

OBJECTIVE: To investigate the method for reconstruction of large tissue defects following surgical resection of advanced oral cancer using pectoralis major myocutaneous flap. METHODS: From 2005 to 2009, 40 patients with advanced oral cancer received extensive surgical resection of oral cancer, and the intraoral defects were reconstructed using pectoralis major myocutaneous flaps. RESULTS: All the flaps survived except one flap with partial necrosis. CONCLUSION: Pectoralis major myocutaneous flap is effective for reconstruction of large tissue defects after resection of advanced oral cancer.

Intracellular CMTM2 negatively regulates human immunodeficiency virus type-1 transcription through targeting the transcription factors AP-1 and CREB.

BACKGROUND: The CKLF-like MARVEL transmembrane domain-containing family (CMTM) is a novel family of proteins linking chemokines and TM4SF. Different members exhibit diverse biological functions. In this study, the effect of intracellular CMTM2 on regulating human immunodeficiency virus type-1 (HIV-1) transcription was evaluated. METHODS: The effects of CMTM2 on regulating full-length HIV-1 provirus and the HIV-1 long terminal repeat (LTR)-directed transcription were assessed by luciferase assay. Transcription factor assays, using the luciferase reporter plasmids of AP-1, CRE, and NF-κB were conducted to explore the signaling pathway(s) that may be regulated by CMTM2. The potential relationship between CMTM2 and the transcription factor AP-1 was further analyzed by Western blotting analyses to investigate the effect of CMTM2 on PMA-induced ERK1/2 phosphorylation. RESULTS: The results from the current study revealed that CMTM2 acts as a negative regulator of HIV-1 transcription. CMTM2 exerted a suppressive action on both full-length HIV-1 provirus and HIV-1 LTR-directed transcription. Transcription factor assays showed that CMTM2 selectively inhibited basal AP-1 and CREB activity. Co-expression of HIV-1 Tat, a potent AP-1 and CREB activator, can not reverse CMTM2-mediated AP-1 and CREB inhibition, suggesting a potent and specific effect of CMTM2 on negatively regulating these two signaling pathways. CONCLUSION: Intracellular CMTM2 can negatively regulate HIV-1 transcription, at least in part, by targeting the AP-1 and CREB pathways. Exploring the mechanisms further may lead to new ways to control HIV-1 replication.

[Expression of MMP1 mRNA in oral squamous cell carcinoma and paired normal tissues].

OBJECTIVE: To investigate the mRNA expression of matrix metalloproteinase 1 (MMP1) gene in oral squamous cell carcinoma (OSCC) and the paired normal tissues. METHODS: The differential expression of MMP1 mRNA between 30 OSCC and paired normal tissues were detected with reverse transcription-PCR (RT-PCR). RESULTS: The relative expression level of MMP1 mRNA in the OSCC tissues showed a 3.26-fold increase in comparison with that in the paired normal tissues (4.06-/+0.52 vs 1.24-/+0.17, P<0.0001). In the 30 OSCC tissues, the relative expression level of MMP1 mRNA was higher in histological grade II/III tissues (4.31-/+0.68) than in grade I (3.87-/+0.57) tissues, higher in OSCC in advanced stages (III/IV) than in tumors in early stages (I/II) (4.18-/+0.67 vs 3.65-/+0.53), and also higher in OSCC with cervical lymph node invasion than in those without cervical lymph node invasion (4.32-/+0.71 vs 3.91-/+0.51), but these differences were not statistically significant (P>0.05). CONCLUSION: MMP1 gene may play a role in local invasion of OSCC, and can serve as a potential biomarker molecule for diagnosis, treatment and prognostic evaluation of OSCC, with also clinical value for OSCC classification.

[Expression of osteopontin mRNA in oral squamous cell carcinoma and normal oral mucosa].

OBJECTIVE: To investigate osteopontin (OPN) mRNA expression in oral squamous cell carcinoma (OSCC) and normal oral mucosa tissues. METHODS: Differential OPN gene expression were detected in 30 cancerous tissues and their paired normal tissues using real-time reverse transcription-PCR (real-time RT-PCR), and the data were analyzed statistically. RESULTS: Real-time RT-PCR results demonstrated that the relative expression level of OPN mRNA in the cancerous tissues were significantly higher than that in paired normal samples (4.17-/+0.51 vs 0.97-/+0.12, P<0.001), showing a 4.3-fold up-regulation. In the 30 OSCC specimens, OPN mRNA expression in the OSCC of histological grades I showed a 3.1-fold down-regulation, significantly lower than the expression in grade II/III tumors (2.16-/+0.17 vs 6.80-/+0.72, P<0.05); its expression was significantly lower in early stage than in advanced stage OSCCs (2.34-/+0.17 vs 4.73-/+0.35, P<0.05). In cases of cervical lymph node metastasis, the expression was significantly higher than that in cases without lymphatic metastasis (6.38-/+0.56 vs 2.89-/+0.32, P<0.05). CONCLUSION: OPN mRNA overexpression may play an important role in OSCC carcinogenesis and can be a potential target for OSCC therapy.

[Single nucleotide polymorphisms of HIV coreceptor CCR5 gene in Chinese Yi ethnic group and its association with HIV infection].

OBJECTIVE: To investigate the single nucleotide polymorphism (SNP) of HIV-1 coreceptor CCR5 gene in Chinese Yi ethnic group and the association between these SNPs and HIV/AIDS. METHODS: Peripheral blood samples of 102 HIV negative persons of Chinese Yi nationality, 87 males amd 15 females, aged 23 (12-37), and 68 HIV carriers, 61 males and 7 females, aged 27 (17-51). The regulatory and structural regions of the HIV coreceptor CCR5 gene were amplified from the genomic DNA by nested PCR, each of the two regions was divided into three gene fragments which were overlapped. High throughput DHPLC was used for screening of unknown mutations in each gene fragment. The PCR products showing different peak traces from wild types in DHPLC were sequenced by forward and reverse primers respectively. The sequences were analyzed with the help of Sequence Navigator software to search for SNP loci. Statistical analysis by SPSS and PPAP softwares were made to study the association between these SNPs and HIV infection. RESULTS: Five SNPs (A77G, G316A, T532C, C921T, and G668A) and a AGA deletion of the 686-688 nucleotides were discovered in the coding region of this gene in Chinese Yi ethnic group. C921T mutation was a nonsense mutation, and the other SNPs (A77G, G316A, T532C, and G668A) are sense mutation, with the amino acid changes of K26R, G106R, C178R, and R223Q. Only the frequency of R223Q allelic gene was high (0.08) but those of the others were low (less than 0.01). There was no significant difference in the allele frequency between the HIV negative and HIV positive groups (all P > 0.05). Five SNP loci (T58934G, G59029A, T59353C, G59402A, and C59653T) were found in the regulatory region of CCR5 gene with high allelic frequencies of 0.1912-0.2941. Between the HIV negative and HIV positive groups, there were no differences in the SNP loc (all P > 0.05). Statistical analysis of the association between the linkage of mutation loci with HIV infection suggested a significant difference in the haplotype frequency of T59353C-G59402A between the HIV negative and HIV positive groups of the Yi population. CONCLUSION: A high throughput screening method of detecting unknown genetic mutation DHPLC can effectively analyze the SNP of CCR5 regulatory and structural regions in Chinese Yi ethnic group.

[Genotyping of HLA-Cw locus in Chinese Yi ethnic group by PCR-SSP].

BACKGROUND: To analyze the genetic polymorphism of HLA-Cw locus in Chinese Yi ethnic group by DNA typing for further study on its association with HIV infection and progression to AIDS. METHODS: A rapid genotyping method for HLA-Cw by PCR-SSP was set up. It combined twenty-six specific primers and one pair of internal control primer to form twenty-four one-step reactions for each sample. Totally 102 unrelated healthy Chinese Yi ethnic individuals were typed. RESULTS: Twelve HLA-Cw alleles were detected in Chinese Yi ethnic group with HLA-Cw*01, Cw*07 and Cw*08 as the most common genes, which accounted for a frequency of 0.333 3, 0.250 0 and 0.176 5 respectively; four kinds of non-serologically defined HLA-Cw genes i.e. Cw*12, Cw*1301, Cw*14 and Cw*15 were found in this population. Hardy-Weinburg test showed that the genotype distribution observed was correspondent with the expected (chi2=65.983 1, df=66, P>0.05). CONCLUSIONS: This study provides the data of HLA-Cw gene frequency in Chinese Yi ethnic group, which may contribute to research on anthropology, disease association and vaccine application. The result also confirmed that PCR-SSP was a reliable and fast method for HLA-Cw genotyping.