Screening of the gene mutation in D-loop region of mitochondrial DNA in oral squamous cell carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the gene mutation in D-loop region of mitochondrial DNA (mtDNA) in oral squamous cell carcinoma (OSCC) tissue and to explore the role of the gene mutation in D-loop region in the OSCC tumorigenesis.</p><p><b>METHODS</b>mtDNA was obtained from cancer, paracancerous and normal mucosa tissues of thirty patients with OSCC. The D-loop regions of mtDNA were amplified with PCR, sequencing and then analyzed by Chromas software and BLAST to identify the mutation site.</p><p><b>RESULTS</b>Mutation in the D-loop region was found in eight cases, with the mutation rate of 27%. There were nine mutations totally, including one point mutation, two base deletions, three insertion mutations, three heterozygous mutations. In these mutations, base deletions were different from each other and heterozygous mutations had no same mutation form, while the three insertion mutations were same, the insertion of base C. One case had T/A heterozygous mutation and base C insertion at the same time.</p><p><b>CONCLUSIONS</b>There were mutations in mtDNA D-loop in OSCC, but the relationship between occurrence of OSCC and mutation of mtDNA needs further study.</p>

Investigation of mRNA expression of collagen genes in oral squamous cell carcinoma and paired normal tissue / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the mRNA expression of collagen genes in oral squamous cell carcinoma (OSCC) and paired normal oral mucosal tissue.</p><p><b>METHODS</b>The differential mRNA expressions of collagen genes between 30 OSCC tissues and the paired normal oral mucosal tissues were detected by RT-PCR.</p><p><b>RESULT</b>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).</p><p><b>CONCLUSION</b>COL1A1 mRNA overexpression may play an important role in the carcinogenesis of OSCC and can be a potential molecular marker of OSCC.</p>

Effects of Podoplanin on cell proliferation and cell cycle in oral leukoplakia cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of Podoplanin on cell proliferation and cell cycle in oral leukoplakia cells.</p><p><b>METHODS</b>Oral leukoplakia cell line (Leuk-1) transfected with pCMV-Podoplanin (A4-1) or pCMV (B4-1) was used in this study. The levels of mRNA and protein of Podoplanin were detected by real-time PCR and Western boltting in A4-1, B4-1 and Leuk-1 cells. The localization of Podoplanin was detected by confocal microscope. Cell growth and proliferation was examined by methyl thiazolyl tetrazolium (MTT) assay and cell cycle was detected by flow cytometry (FCM).</p><p><b>RESULTS</b>The value of Podoplanin mRNA level in A4-1 cells was 0.022, which was significantly higher than the values in B4-1 (0.001) and Leuk-1 cells (0.002), P < 0.05. The gray scale of Podoplanin protein in A4-1 cells was significantly higher than in the control groups (P < 0.05). The expression of Podoplanin was observed in cell plasm and membrane of A4-1, B4-1 and Leuk-1 cells. But the expression level of Podoplanin in A4-1 cells was higher than in control groups. A4-1 cells grew faster than Leuk-1 cells. The proliferation rate after 3 days of culture in A4-1 cells was 40.4% higher than that in B4-1 cells (P < 0.05). The G₂-M phase (24.89 +/- 4.55)% and PI (0.57 +/- 0.06) of A4-1 cells were significantly higher than that in B4-1 cells [(4.13 +/- 5.24)%, (0.41 +/- 0.04)] and Leuk-1 cells [(4.69 +/- 7.42)%, (0.40 +/- 0.02)], P < 0.05.</p><p><b>CONCLUSIONS</b>Over expression of Podoplanin accelerated the growth and proliferation of oral leukoplakia cells. Podoplanin may be one of key genes in the malignant transformation of oral leukoplakia.</p>

Inflammatory factors promote oral squamous cell carcinoma cells metastasis, via nuclear factor kappa B signal pathway in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the roles of inflammatory factors and nuclear factor kappa B (NF-kappaB) signal pathway in metastasis of oral squamous cell carcinoma.</p><p><b>METHODS</b>The oral squamous cell carcinoma cell lines with highly metastasis potential (Tb) and lower metastasis potential (Tca8113) were used in this study. The levels of NF-kappaB activity in oral squamous cell carcinoma cell lines were determined by Western blotting and luciferase reporter assay. pBalphabe-IkappaBalpha-SR expression vector or NF-kappaB inhibitor pyrolidinedithiocarbamate (PDTC) was used to inhibit NF-kappaB, and cell migration was examined by transwell assay. The secretion of tumor necrosis factor-alpha (TNF-alpha), IL-1alpha, IL-6, IL-8 and GM-CSF proinflammatory cytokines was determined by ELISA when Tb cells were transfected with pBalphabe-SR-IkappaBalpha or treated with PDTC.</p><p><b>RESULTS</b>Western blotting showed that the levels of phosphorIkappaBalpha and phosphor-p65 were highly expressed in Tb cells. Tb cells had high level of constitutive NF-kappaB activity and were more sensitive to TNF-alpha. The migration of highly metastatic Tb cells, either transfected with dominant-negative mutant inhibitor pBalphabe-SR-IkappaBalpha or treated with PDTC, was suppressed when determined by transwell assay. The secretion of proinflammatory cytokines, including TNF-alpha, IL-1alpha, IL-6, IL-8 and granulocyte-macrophage colony stimulating factor (GM-CSF), was inhibited by pBalphabe-SR-IkappaBalpha transfection or PDTC treatment.</p><p><b>CONCLUSIONS</b>The inflammatory factors such as TNF-alpha could promote oral squamous cell carcinoma cell metastasis via NF-kappaB signal pathway.</p>

Effect of Matrine on cell cycle and human telomerase reverse transcriptase of human salivary adenoid cystic carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of the traditional Chinese medicine Matrine on cell cycle and human telomerase reverse transcriptase (hTERT) of human ACC-M cell lines.</p><p><b>METHODS</b>Different concentrations of Matrine were used in the medium of ACC-M cells. Change of cell cycle were detected by flow cytometry after ACC-M cell were cultivated with different concentrations Matrine (0.25, 0.50, 0.75, 1.00 mg x mL(-1)). Expression of hTERT was investigated by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescene and flow cytometry quantitative analysis.</p><p><b>RESULTS</b>Matrine caused obviously the GdG1 phase block and inhibited proliferation of ACC-M cells. At same time, this effect was positive correlation to Matrine concentration and treat time. Matrine can inhibit the expression of hTERT mRNA and protein.</p><p><b>CONCLUSION</b>Matrine can obviously inhibit cell cycle and down-regulate expression of hTERT. Inhibition of cell cycle is possible correlation with down-regulation expression of hTERT.</p>

Benzo (a) pyrene induced tumorigenesity of human immortalized oral epithelial cells: transcription profiling / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The present study was designed to examine and analyze the global gene expression changes during the tumorigenesis of a human immortalized oral epithelial cell line, and search for the possible genes that may play a role in the carcinogenesis of oral cancer associated with benzo (a) pyrene.</p><p><b>METHODS</b>The human immortalized oral epithelial cells, which have been established through transfection of E6/E7 genes of human papillomavirus type 16 and proved to be non-tumorigenic in nude mice, were treated with benzo (a) pyrene. Tumorigenicity of the treated cells were examined through nude mice subcutaneous injection. The global gene expression profiles of immortalized cells and the tumorigenic cells were acquired through hybridization of a microarray of Affymetrix U133 plus 2.0. The data were analyzed using Spring 7.0 software and treated statistically using one-way analysis of variance (ANOVA). The differentially expressed genes were classified using a Venn diagram and annotated with gene ontology. Several highlighted genes were validated in cells using a real-time polymerase chain reaction.</p><p><b>RESULTS</b>There were 883 differentially expressed genes during the tumorigenesis and most of them changed expression in the early stage of tumorigenesis. These genes mainly involved in macromolecule metabolism and signal transduction, possessed the molecular function of transition metal ion binding, nucleotide binding and kinase activity; their protein products were mainly integral to membranes or localized in the nucleus and cytoskeleton. The expression patterns of IGFBP3, S100A8, MAP2K, KRT6B, GDF15, MET were validated in cells using a real-time polymerase chain reaction; the expression of IGFBP3 was further validated in clinical oral cancer specimens.</p><p><b>CONCLUSIONS</b>This study provides the global transcription profiling associated with the tumorigenesis of oral epithelial cells exposed to benzo (a) pyrene; IGFBP3 may play a potential role in the initiation of oral cancer related to benzo (a) pyrene exposure.</p>

Expression of osteopontin mRNA in oral squamous cell carcinoma and normal oral mucosa / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate osteopontin (OPN) mRNA expression in oral squamous cell carcinoma (OSCC) and normal oral mucosa tissues.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>OPN mRNA overexpression may play an important role in OSCC carcinogenesis and can be a potential target for OSCC therapy.</p>

Expression of MMP1 mRNA in oral squamous cell carcinoma and paired normal tissues / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the mRNA expression of matrix metalloproteinase 1 (MMP1) gene in oral squamous cell carcinoma (OSCC) and the paired normal tissues.</p><p><b>METHODS</b>The differential expression of MMP1 mRNA between 30 OSCC and paired normal tissues were detected with reverse transcription-PCR (RT-PCR).</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>

A new mutation of PTCH gene in a Chinese family with nevoid basal cell carcinoma syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Nevoid basal cell carcinoma syndrome (NBCCS) is a rare autosomal dominant disease characterized by a combination of development anomalies and a predisposition to tumour formation. Mutation of patched gene (PTCH), considered the molecular defect of NBCCS, in a Chinese NBCCS family was investigated in this study.</p><p><b>METHODS</b>Genomic DNA was isolated from blood samples of all 12 members of this family. The mutated PTCH gene was screened by polymerase chain reaction amplification and direct sequencing.</p><p><b>RESULTS</b>A new mutation of 3 bp (GAT deletion) was found in all seven affected members of this family. This mutation caused one aspartate deletion in the fourth transmembrane domain of the PTCH protein located within the sterol sensing domain (SSD). This deletion was not found in any unaffected members of this family nor in 200 control samples.</p><p><b>CONCLUSIONS</b>Our findings suggest that one 3-bp deletion in PTCH gene was the cause of nevoid basal cell carcinoma in a Chinese family through affecting the conformation and function of PTCH protein.</p>

Establishment and identification of biological characteristics of rat monoclone cell line Rca-B / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To establish a monoclone cell line of squamous cell carcinoma (SCC) in rat buccal mucosa and to study its biological characteristics.</p><p><b>METHODS</b>SCC in rat oral mucosa was induced by adding 4-nitroquinoline-1-oxide (4NQO) into the SD rats' drinking water, and the cancer cells were then cultured to obtain mixed cells in vitro. The mixed tumor cells were purified by mono cell cloning method. The biological characteristics of the cells were studied by microscope and electronic microscope observation, chromosome analysis, Methyl thiazolyl tetrazolium (MTT) test, flow cytometry assay and immunohistochemistry staining. Hypodermic inoculations of the cells in nude mice and injection of the cells by nude mice tail veins were performed to observe the tumor formation and long distance metastasis.</p><p><b>RESULTS</b>The morphology proved that the cell line was squamous cell carcinoma cells, which were cultured from one cell. The population doubling time for passage 65 cells was 25.44 hours. The cells in S-phase accounted for 20.13% of the cell cycle. The chromosome modal number was 84. All the cells expressed the proteins of cytokeratin and vimentin. The xenograft rate and the tumor metastatic rate to the lung were 100% in nu/nu BALB/C mice, but the homograft rate was zero in SD Rats.</p><p><b>CONCLUSIONS</b>Rca-B was a typical oral squamous cell carcinoma cell line derived from Sprague-Dawley rat buccal mucosa carcinoma, and the cell line has high metastatic potential and its biological characteristics were well ascertained.</p>

A20 inhibits human salivary adenoid cystic carcinoma cells invasion via blocking nuclear factor-kappaB activation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A20, also known as tumor necrosis factor alpha induced protein 3 (TNFaip3), is a cytoplasmic zinc finger protein that inhibits nuclear factor kappa-B (NF-kappaB) activity and prevents tumor necrosis factor (TNF)-mediated programmed cell death. NF-kappaB is a transcription factor that regulates expression of genes involved in cell proliferation, cell survival and anti-apoptosis. Several studies have implicated that the NF-kappaB signal pathway is associated with angiogenesis and clinico-pathological process of adenoid cystic carcinoma (ACC) of the salivary glands.</p><p><b>METHODS</b>The ability of overexpression of A20 to influence the biological behavior and invasion of ACC cells was examined. The cells were stably transfected with full-length A20 cDNA. Stable gene transfer was verified by realtime-polymerase chain reaction (PCR) and Western blot analysis. The change of cell biological behavior was examined by methyl thiazolyl tetrazolium (MTT) and NF-kappaB luciferase reporter assay and the invasion of the cells was examined by a Matrigel invasion chamber.</p><p><b>RESULTS</b>pEGPFN3-A20 gene was stably transferred into ACC-2 cells and overexpressed. When cells were treated with TNFalpha, the NF-kappaB activity of ACC-2-A20 cells could be down-regulated about 46.32% in contrast to ACC-2-GFP cells (P < 0.05). A20 potently inhibited growth of A20 transfectant ACC-2-A20 compared with control vector transfected groups and the ACC-2 empty control group (P < 0.05). The ACC-2-A20 cells showed significantly reduced ability to invade through Matrigei-coated filters compared to ACC-2-GFP and ACC-2 cells. The inhibition rate was up to 71.05% (P < 0.05).</p><p><b>CONCLUSIONS</b>A20 gene transfer is associated with decreased tumor invasion, in part via the down-regulation of NF-kappaB expression, providing evidence for a potential application of A20 in designing a treatment modality for salivary gland cancers such as ACC.</p>

In vitro effect of PTEN gene transfection on growth of human colon cancer cell lines / 上海第二医科大学学报

Objective To investigate the in vitro effect of wild-type PTEN gene transfection on the growth of human colon cancer cell lines. Methods With liposome transduction technique,the retrovirus vector pBp-PTEN,which contains wild-type PTEN gene segment and pBabe-puro,was transfected into Lovo,one of the PTEN-deficient human colorectal carcinoma cell lines.After identification by Western blotting,cell growth,cell cycle and apoptosis before and after transfection were studied. Results It was indicated by the cell growth curve that after transfection the curve of the transfection group exhibited a mild tendency with no obvious logarithmic growth phase,and the growth velocity was significantly lower than that of the control group(P

The establishment of a carcinogenesis model of oral squamous cell carcinoma in vitro / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To transform HPV E6/E7 immortalized human oral epithelial cell (HIOEC) line cells by benzo(a)pyrene [B(a)P] in vitro, and to establish a carcinogenesis model of oral squamous cell carcinoma.</p><p><b>METHODS</b>HIOEC cells were treated with 0.1 mg/L-1.2 mg/L B(a)P for 6 months. The cells were cloned at 18th passage, and then the culture medium was changed into DMEM containing 10% FBS at 21th passage. Cells were cultured in vitro for half and one year and the cell line was named HIOEC-B(a)P. The morphological changes of the cells were observed with differential interference contrast microscope and HE staining. The soft agar colony forming ability and tumorigenicity of the cells in nude mice were identified to confirm the malignant characteristics of HIOEC-B(a)P cells.</p><p><b>RESULTS</b>(1) After HIOEC cells were treated with B(a)P for 6 months, HIOEC-B(a)P cells could grow well in DMEM medium containing 10% FBS and physical concentration of calcium. (2) When HIOEC cells were treated with chemical carcinogens, the morphology of the cells was changed. Cells showed the character of polygon epithelial cells with much atypical mitosis. (3) The 93th passage of HIOEC-B(a)P cells had soft agar colony formation ability. (4) The 55th passage of HIOEC-B(a)P cells could develop parakeratosis mass. The 69th passage of HIOEC-B(a)P cells could develop typical well-differentiated squamous cell carcinoma. The 74th and the 96th HIOEC-B(a)P cells developed I-II grade squamous cell carcinoma-like clinical lesions in nude mice.</p><p><b>CONCLUSIONS</b>B(a)P may induce HIOEC cells to be oral squamous cell carcinoma (OSCC) carcinogenetic cells. It will provide a multiple factors, multistage carcinogenesis model of OSCC for the further research.</p>

Inhibition effect on Tca8113/CDDP by antisense oligonucleotides of cyclin D1 combined with cis diamminedichloroplatinum / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To clarify the relationship between cyclin D1 and cisplatin resistance of Tca8113/cis diamminedichloroplatinum (CDDP) in vitro and in vivo.</p><p><b>METHODS</b>We applied the transfection method with plasmids pcDNA3.1-antisense-cyclin D1 by Lipofectamine 2000. Tca8113/CDDP cells were used as control. MTT assay was used to identify the proliferation and sensibility of those cells to cisplatin. Subsequently, 18 nude mice were subcutaneously injected by those cells and divided into 3 groups with 6 mice in each group. Every mouse was treated by cisplatin with 5 mg . kg(-1) . d(-1) for 5 days. The sizes of tumor were measured every other day and were described with the growth curves. After 20 days, tumors were anatomized and weighed. The tumor inhibition ratios were calculated and the HE slides were observed to determine the cell sensibility to cisplatin.</p><p><b>RESULTS</b>The transfected cells with pcDNA3.1-antisense-cyclin D1grew more slowly than other cells and showed higher sensibility to cisplatin in vitro. The tumors developed by cells with pcDNA3.1-antisense-cyclin D1 were smaller than the The tumor inhibition ratio was 74% (P < 0.05). The necrosis area was larger in the tumors developed by the transfected cells with pcDNA3.1-antisense-cyclin D1 than other groups.</p><p><b>CONCLUSIONS</b>Antisense oligonucleotides of cyclin D1 can improve the sensibility of Tca8113/CDDP cells to cisplatin and inhibit the growth of tumors.</p>

Selection and quantitative detection of target genes in oral squamous cell carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To select and identify the target genes related to oral squamous cell carcinoma (OSCC) and provide target genes for designing oligo-nucleotide functional microarray of OSCC.</p><p><b>METHODS</b>Genes possibly related to oral squamous cell carcinoma were selected from the 5 years' published data of differently expressed profiles with microarray testing in OSCC. Then mRNA expression of selected genes were evaluated by real time quantitative polymerase chain reaction (RT-PCR) in 22 cases of OSCC, including tumor tissues and paried normal mucosas and quantified according to an internal control GAPDH.</p><p><b>RESULTS</b>Eight genes were tested. The overexpression of SPARC, PDGF-A, SERPINE1, TGF-beta(1) and VEGF-C genes were measured in 16, 18, 16, 20, 18 cases of tumor specimens, respectively. The expression of CK15 gene was lower than that of its normal tissue. There were overexpression of CCND1, BIRC3 in tumor tissues, but there was no significant difference of CCND1 and BIRC3 expression between tumor tissue and normal tissue (P > 0.05).</p><p><b>CONCLUSIONS</b>SPARC, PDGF-A, SERPINE1, TGF-beta(1), VEGF-C and CK15 genes were closely related to tumor progress of OSCC. They can be used as the target genes for designing oligo-nucleotide functional microarray of OSCC.</p>

In vitro gene silencing by siRNA of cyclin D1 on tongue squamous cell carcinoma cell line resistant to cisplatin / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To interfere in the Tca8113-CDDP cell line with siRNA of cyclin D1 and to investigate time and dose dependent gene silencing effect of siRNA of cyclin D1.</p><p><b>METHODS</b>siRNA of cyclin D1 was transfected into Tca8113-CDDP cells Fluorescent CY3 dye labeled siRNA GAPDH was used as the control. The transient transfecting efficiency was examined at 4, 24, 48 and 72 h. The relative quantity of the target RNA of cyclin D1 was analyzed with SYBR Green fluorescent dye kit by the Real-time PCR assay. The protein level of cyclin D1 was examined with Western blot. The changes of cisplatin sensitivity after treatment with siRNA cyclin D1 were examined with methyl thiazolyl tetrazolium (MTT) assay.</p><p><b>RESULTS</b>The optimized transfecting efficiency with CY3 labeled siRNA GAPDH in Tca8113-CDDP cells was over 90%. The silencing rate of cyclin D1 siRNA was 81.6% at 24 h, 80.7% at 48 h and 94.3% at 72 h. Dose-dependent manner of gene silencing effect was observed when the siRNA concentration was lower than 100 nmol/L, however, gene silencing effect reached its platform when the concentration was higher than 100 nmol/L. The protein levels of cyclin D1 at 24, 48 and 72 h after transfection decreased significantly, and so did the growth of cells. Inhibition rates of cell growth induced by cisplatin after administration with or without cyclin D1 siRNA were 58.4% and 34.8%, respectively.</p><p><b>CONCLUSIONS</b>Chemical synthesized cyclin D1 siRNA effectively silenced the expression of cyclin D1 gene in Tca8113-CDDP cells in vitro, with a time- and dose-dependent manner and target gene silence in cells increased its sensitivity to cisplatin.</p>

Modulation of Chinese regimen granules of Shenyang on peripheral blood lymphocyte subsets of SD rats with SCC of tongue / 中华口腔医学杂志

<p><b>OBJECTIVE</b>Prospective research demonstrated that Chinese regimen granules of Shenyang could prolong survival time and improve survival rate of patients with oral squamous cell carcinoma (SCC). But the mechanism was not clear. The purpose of this study was to investigate Shenyang's effect on peripheral blood lymphocyte subsets of SD rats with SCC of tongue and explore immunological mechanism.</p><p><b>METHODS</b>Among 80 SD rats fed by 0.002% 4-nitroquinoline-1-oxide (4NQO) drinking water for 36 weeks, 61 rats with SCC of tongue had been found and were randomly divided into 4 groups, namely Shenyang A, Shenyang B, positive and blank control groups. Before and after high and normal dosage of Shenyang, acanthopanax senticoside and water had been given for 15 days respectively, peripheral blood lymphocyte subsets were detected with flow cytometry. The data were statistically analyzed with paired t Test.</p><p><b>RESULTS</b>Percentage of CD3+ CD4+ T cell and CD3-CD161a+ NK cell, ratio of CD4+/CD8+ were increased. Percentage of CD3+CD8+ T cell was decreased, and the effect was better than that of acanthopanax senticoside in improving the percentage of CD3-CD161a+ NK cell.</p><p><b>CONCLUSION</b>Among anti-tumor mechanisms of Shenyang it is that corrects disorder of lymphocyte subsets and increases percentage of CD3-CD161a+ NK cell.</p>

The effect of intraperitoneal heparin on distribution of ACC-M in lung and liver / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of intraperitoneal heparin on the adhesion of highly lung metastasic adenoid cystic carcinoma cell line (ACC-M) in lung.</p><p><b>METHODS</b>3HTdR labeled ACC-M cells were injected by intravenous infusion after intraperitoneal injection with 200 units heparin. 4 mice of each group were killed at 2 h, 6 h and 18 h after infusion. The relative radioactivity (CPM) in lung and liver was detected.</p><p><b>RESULTS</b>3H-activity per gram in lung of heparin group was lower than control at the same time. There was significant difference between the two groups (P < 0.001). There was no difference between the two groups in liver (P > 0.05).</p><p><b>CONCLUSION</b>The results of this study suggest that the highly lung metastasis characteristic of ACC-M may be partially inhabited by the use of intraperitoneal heparin.</p>

Construction and confirmation of the fused plasmid with Rb bait gene in yeast two-hybrid system / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To screen and clone the novel genes related to cellular proliferation of oral squamous cell carcinoma.</p><p><b>METHODS</b>We selected the Rb gene as the bait protein gene to construct the fusion bait plasmid of yeast two-hybrid. The whole code sequence of Rb gene was acquired by digestion with restricted enzyme EcoRI and BamH1 and reclaimed from its original vector pGBT9-pRb. After being confirmed by electrophoresis, the Rb gene was cloned into the MCS of the plasmid pGBKT7 to construct a recombined plasmid pGBKT7-pRb and the sequence of the recombined plasmid was detected in company. According to the protocol of yeast two hybrid system III, the competent Y187 yeast was prepared, and transformed with recombined plasmid pGBKT7-pRb. Following that, the toxicity and transcriptional activation of this recombined plasmid pGBKT7-pRb in Y187 yeast were tested.</p><p><b>RESULTS</b>The sequence of the recombined plasmid was correct compared with the sequence provided in Genbank. The protein could be correctly synthesized in vitro, and no self-activating transcriptional activation and toxicity was observed in Y187 yeast.</p><p><b>CONCLUSIONS</b>The construction of the recombined plasmid was capable to be used as the fusion bait plasmid in yeast two-hybrid system III, and the recombined Rb-protein could be used as the bait protein successfully.</p>

Chemotherapy with higher or lower dose of teniposide combined with cisplatin and pingyangmycin for oral squamous cell carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To compare the clinical efficacy and toxicity of teniposide (VM26) of higher dose with those of lower dose, both combined with cisplatin (CDDP) and pingyangmycin (PYM), in the treatment of patients with squamous cell carcinoma of oral and maxillofacial region (SCCOMR).</p><p><b>METHODS</b>Sixty-five patients with SCCOMR entered into this study prospectively. Thirty-three patients were treated with higher dose of VM26 (total dose was 320 mg) combined with CDDP and PYM (PTP1), the other thirty-two patients were treated with lower dose (total dose was 158 mg) of VM26 combined with CDDP and PYM (PTP2).</p><p><b>RESULTS</b>Thirty-three patients received a total of 38 cycles of PTP1. The overall response rate was 81.82% (27/33). Thirty-two patients received a total of 36 cycles of PTP2 and showed overall response rate by 81.25% (26/32). There was no significant difference between PTP1 and PTP2 groups in response rate (P > 0.05). But the blood toxicity was more severe in PTP1 group than in PTP2 group (P < 0.01). Bone marrow depression rate (1-4 stage) was 48.48% in PTP1 group versus 25.00% in the other group.</p><p><b>CONCLUSIONS</b>A high response rate of 81.25% and relatively slighter adverse events could be obtained for lower dose of VM26 combined with CDDP and PYM (PTP2). So, the chemotherapy schedule, PTP2, a novel teniposide based regimen in SCCOMR could be employed and spread in clinical practice.</p>