Effects of silencing inhibitor of DNA binding-1 gene on the growth and invasiveness of adenoid cystic carcinoma cells / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the role of inhibitor of DNA binding-1 (Id-1) gene in adenoid cystic carcinoma cell growth and invasion behavior.</p><p><b>METHODS</b>With salivary adenoid cystic carcinoma cell lines ACC-M and ACC-2, dedected Id-1 gene expression was screened with immunofluorescence assay. After Id-1 mRNA knocking-down using small interfering RNA, RT-PCR and Western blot were used to detect the different expressions before and after interference, and the growth of cells before and after interference was deceted using the MTT assay, and the cell invasion ability was checked with the use of Transwell chamber assay.</p><p><b>RESULTS</b>Id-1 were both expressed in the ACC-M and ACC-2, and the expression in ACC-M was higher than that in ACC-2. After Id-1 RNA interference, the growth and invasiveness of ACC-M and ACC-2 were inhibited with the restrained degree in ACC-M much stronger than that in the ACC-2.</p><p><b>CONCLUSION</b>In view of the important role of Id-1 in the behavior of growth and invasion in ACC cell, interfering the expression of Id-1 gene is expected to be a novel and effective means for the treatment of adenoid cystic carcinoma.</p>

The expression of interleukin-10 mRNA in gingival lesion of different clinical states in patients with adult periodontitis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression of interleukin-10 (IL-10) mRNA in gingival tissue of active and stable stage in patients with adult periodontitis.</p><p><b>METHODS</b>12 patients with acute abscesses of the periodontium, 12 patients after periodontal initial treatment and 6 periodontal healthy patients having extraction of impacted wisdom tooth were randomly divided into group A (active stage group), group B (stable stage group) and the control group. Biopsies of gingival tissues were collected from every subject of three groups. Technique of in situ hybridization was applied to observe the expression of IL-10 mRNA in the biopsies from three groups semi-quantitatively.</p><p><b>RESULTS</b>IL-10 mRNA was positively expressed in lymphocytes, macrophages and fibroblasts. The quantity of IL-10 mRNA of group A was the lowest in the three groups and was significantly lower than that of control group and group B respectively (P < 0.01). The quantity of IL-10 mRNA of group B was the highest in the three groups and was significantly higher compared with the control group and group A (P < 0.01).</p><p><b>CONCLUSION</b>The quantities of IL-10 mRNA expression are closely related with various clinical states of periodontitis.</p>

Morphologic observation of oral cancer cells cocultured with mesenchymal cells in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the morphologic and growing alterations of oral cancer cell line Tca8113 before and after cocultured with tumor stromal fibroblasts (TSF) and normal stromal fibroblasts (NSF) respectively, and evaluate the influence of mesenchymal cells on tumor cells.</p><p><b>METHODS</b>TSF and NSF were isolated and cultured. To observe the morphologic change of Tca8113 cells after cocultured with TSF and NSF respectively.</p><p><b>RESULTS</b>When cocultured with NSF, the Tca8113 cells proliferated as rapidly as monocultured to form colonies, while the NSF proliferated slowly to form pieces and then joined each other to form network. The NSF network segmented and surrounded the colonies of cancer cells so that the cancer cells shrank, turn round, broke away from the bottom and floated into the medium. The cancer cells proliferated actively but they were elbow out entirely in the end. TSF proliferated slowly when cocultured with cancer cells, projected several branched protrusions. The cancer cells proliferated along the two sides of protrusions of TSF, or projected short protrusions to connect the body or protrusions of TSF, and overlaid the protrusions gradually, finally, cover the body. In the end, TSF melt away, and the cancer cells took on the figure of TSF.</p><p><b>CONCLUSION</b>The results do suggest that, oral cancer cell line Tca8113 are restrained when coculture with NSF, but are promoted when with TSF.</p>

Effect of curcumin on invasion and migration of tongue squamous cell carcinoma cell line Tca8113 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the effect of curcumin on invasion and migration of the tongue squamous cell line Tca8113.</p><p><b>METHODS</b>Tca8113 cells were treated with curcumin (0 - 100 micromol/L) for 24 h and the conditional medium was collected. The gelatinases - matrix metalloproteinase -2 and -9 (MMP-2, -9) in the conditional medium were detected by gelatin zymography. The cell invasion and migration model in vitro was conducted using transwell chamber with or without matrigel. Using this model, the effects of 50 micromol/L curcumin on invasion and migration of Tca8113 were detected.</p><p><b>RESULTS</b>Curcumin reduced the activities of MMP-2 and MMP-9 on a dose-dependent manner. Curcumin can suppress cell invasion and migration significantly (P <0.01).</p><p><b>CONCLUSIONS</b>Curcumin can suppress Tca8113 invasion and migration by reducing the activities of MMP-2 and MMP-9.</p>

Effect of hypoxia inducible factor-1 alpha on vascular endothelial growth factor expression in human tongue squamous carcinoma cells (Tca8113) under hypoxia / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of hypoxia inducible factor-1 alpha (HIF-1 alpha) on vascular endothelial growth factor (VEGF) expression in Tca8113 cells under hypoxia.</p><p><b>METHODS</b>The expression of the mRNA of HIF-1 alpha and VEGF in Tca8113 cells was examined by RT-PCR technique at different culture times (1/2 h, 1 h, 3 h, 6 h, 12 h, 24 h) under normoxic and hypoxic conditions.</p><p><b>RESULTS</b>The expression of HIF-1 alpha under hypoxia showed the trend of increasing first and then decreasing, and was higher than that of the control (normoxic group) at 6h and 12 h (P < 0.05). The expression of VEGF under hypoxia was higher than that of the control group at 1/2 h, 1 h, 3 h, 12 h, 24 h (P < 0.05). The expression of hypoxia-induced VEGF mRNA increased with the increased expression of HIF-1 alpha mRNA in the cell lines tested at the initial stage of hypoxia. But no statistical significant association was observed between HIF-1 alpha and VEGF expression within 24 h under hypoxia (rs = 0.5750, P > .005).</p><p><b>CONCLUSIONS</b>The increased expression of VEGF in Tca8113 cells might be mediated by multiple factors, including HIF-1 alpha.</p>

Effect of bone morphogenetic proteins-2 on mandibular distraction osteogenesis in rabbits / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the Effect of recombinant adenovirus vectors containing human Bone morphogenetic proteins-2 (Ad-hBMP-2) on the for mandibular distraction osteogenesis (DO) in rabbits.</p><p><b>METHODS</b>Twenty-four New Zealand white rabbits were randomly divided into experimental group, and control group and underwent mandibular distraction osteogenesis. After 5 days latency, the distracters were activated at a speed of 0.5 mm every 12 hours for 7 days, then on the first day in the consolidation period, the distraction gaps of experimental group were injected with 0.2 ml Ad-hBMP2 10(12) pfu/L, while the animals of control group were injected with 0.2 ml Ad-EGFP 10(12) pfu/L. At the 7 th and 28 th day of consolidation period, specimens were obtained, X-ray and histomorphology were performed. The bone density and the quantity of new bone formation in the distraction gaps were observed and compared between the two groups at different consolidation period.</p><p><b>RESULTS</b>Ad-hBMP-2 treated specimens demonstrated an increased amount of new bone formation</p><p><b>CONCLUSIONS</b>Adenovirally-mediated delivery of BMP-2 can locally increase bone deposition during DO, which may potentially shorten the consolidation period.</p>

Experimental study of mandibular periosteal distraction in rabbits / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate a novel technique for new bone formation--periosteal distraction osteogenesis.</p><p><b>METHODS</b>A custom made periosteal distraction device was fixed to bilateral surface of the mandible in three rabbits. Periosteal distraction was performed on the left side of the mandible, the right side of the mandible served as the control. The animals were sacrificed at the end of distraction process. All the specimens were X-rayed and histologically examinated.</p><p><b>RESULTS</b>All three animals survived with no obvious complications. Both in mass specimens and X-rays, there showed new bone formation on the distracted side of the mandible. In histological examinations, there was osteoblast-like cell infiltration and bone tissue formation in the distracted area.</p><p><b>CONCLUSION</b>Periosteal distraction osteogenesis can provide a novel technique for the repair of bone defects.</p>

Clinical study of oral and maxillofacial malignancies associated with multiple primary malignant neoplasms / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the clinical characteristics, diagnosis and treatment strategy of oral and maxillofacial malignancies in multiple primary malignant neoplasms (MPMNs).</p><p><b>METHODS</b>21 cases of oral and maxillofacial malignancies associated with MPMNs admitted to our hospital between 1985 and 2000 were studied respectively.</p><p><b>RESULTS</b>There were 44 malignant cases in the 21 MPMNs patients. Among the 44 cases, there were 24 cases in alimentary and respiratory tract such as oral, pharynx, esophagus, stomach and lung, and 10 cases in salivary gland, breast and female reproductive system. There were 25 cases malignant neoplasms in oral and maxillofacial region where squamous cell carcinoma was the most common pathologic type, secondly adenoid cystic carcinoma. In oral and maxillofacial region, MPMNs were often found in tongue, parotid and submandibular gland, buccal mucosa and gingival.</p><p><b>CONCLUSION</b>Tongue and salivary gland were the common locations with MPMNs, and they were closely associated with alimentary and respiratory tract. Patients with malignancies of oral and maxillofacial region associated with MPMNs must be submitted to a long-term and careful follow-up. For female patients, breast and female reproductive system should be examined specially. Regular follow-up, early detection, early diagnosis, active and effective treatment can help to improve the survival quality of MPMNs patients.</p>

Study of induction of tumor specific cytotoxic T lymphocyte by using tumor-derived exosome / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate whether the exosomes derived from Tca8113 could induce production of tumor-specific T cells when pulsed onto dendritic cells.</p><p><b>METHODS</b>Tca8113 cell was cultured with RPIM1640, isolated and purified the tumor-derived exosomes from the culture supernatants by ultrafiltration with Millipore centrifual filter devices; frozen and thawed Tca8113 cells to get frozen tumor antigens (FTA). The exosomes and FTA was pulsed onto DC generated from normal human peripheral blood mononuclear cell (PBMC) in vitro. The DC pulsed with FAP or exosomes cocultured with the peripheral blood lymphocytes to transform T cell into specific CTL. To observe the killing and wounding activity of CTL, the CTL and Tca8113 cells were mixed at a ratio of 20 to 1, SPCA-1 cells and 95-D cells was evaluated as control group.</p><p><b>RESULTS</b>The CTL induced by DC pulsed with FAP or exosomes had significant activity killing Tca8113 (P < 0.01); Moreover the CTL induced by DC pulsed with exosomes could also kill SPCA-1 cells (P < 0.05), but the CTL induced by DC pulsed with FTA had not this function.</p><p><b>CONCLUSION</b>Exosomes derived from tumour accumulate in culture supernatants. Exosomes are a natural and new source of tumour-rejection antigens, opening up new avenues for immunotherapy against oral cancers. The exosome-specific CTL could kill another kind of tumor, so tumor-derived exosomes may contain shared tumor-rejection antigens.</p>

Expression of stromal CD34 and alpha-smooth muscle actin in oral invasive cancer / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To observe the expression of stromal CD34 and alpha-smooth muscle actin (alpha-SMA) in oral invasive cancer.</p><p><b>METHODS</b>The distribution and expression of stromal CD34 and alpha-SMA in 60 patients with invasive oral squamous cell cancer and 20 resections with tumor-free margins from 60 patients with invasive oral squamous cell cancer were examined by immunohistochemistry SP method.</p><p><b>RESULTS</b>Twenty cases of resections of mucosa with tumor-free margins exhibited CD34 expression in fibrocytes cytoplasm in the vicinity of vessels, mucosa and submucosa but were free of alpha-SMA expression, only the walls of muscularized vessels stained positive for alpha-SMA in the stroma. Sixty invasive oral squamous cell carcinomas were free of stromal CD34 expression, only the endothelia of small vessels stained positive for CD34, of which 53 invasive oral squamous cell carcinomas expressed stromal alpha-SMA in myofibroblasts cytoplasm.</p><p><b>CONCLUSIONS</b>The detection of CD34 and alpha-SMA is an adjunctive tool in judging oral cancer invasion in small oral mucosa biopsy specimens based on the absence of CD34 expression paralleled by the gain of alpha-SMA in the stroma of oral invasive cancer.</p>

Experimental study of facial nerve regeneration through polytetraflouroethylene tube with Schwann cells self-planted / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To observe the effect of using polytetraflouroethylene (e-PTFE) tube with self-Schwann cells implanted to repair facial nerve defect.</p><p><b>METHODS</b>Enzymatic digest method was used to get pure Schwann cells in short time. The e-PTFE membrane tube was used to bridge the 1.0 cm defect of facial nerve and pure self-Schwann cells were injected into the tube. As control group, the e-PTFE tube without self-Schwann cells was used in the same way. Electric physiological and histological examinations were taken in different times.</p><p><b>RESULTS</b>The effect of nerve regeneration of the experimental group was better than control group at any time. The nerve conduction velocity of the experimental group was 29.70 m/s in the 16th week, while the control groups was 23.00 m/s respectively at the same time.</p><p><b>CONCLUSION</b>It is possible to obtain sufficient active Schwann cells by enzymatic digest method. Using e-PTFE tube to bridge the defect of facial nerve with self-Schwann cells implanted can get effect of nerve regeneration.</p>

Mechanisms of p16 gene inactivation salivary adenoid cystic carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the mechanism of p16 gene inactivation in salivary adenoid cystic carcinoma.</p><p><b>METHODS</b>53 cases of freshly excised salivary adenoid cystic carcinomas were studied. Polymerase chain reaction (PCR) and single-stranded conformation polymorphism analysis of polymerase chain reaction products (PCR-SSCP) were used to detect deletion and mutation of p16 gene in salivary adenoid cystic carcinomas. Methylation specific PCR (MSP) was used to detect the methylation status of p16 gene.</p><p><b>RESULTS</b>The homozygous deletion, mutation and hypermethylation of p16 gene were noted in 16 cases (30.2%), 4 cases (7.5%) and 26 cases (49.1%) respectively in 53 cases of salivary adenoid cystic carcinomas.</p><p><b>CONCLUSION</b>The main inactivation mechanisms of p16 gene in salivary adenoid cystic carcinoma were hypermethylation and homozygous deletion. The mutation p16 gene was rare in salivary adenoid cystic carcinoma.</p>

Study of the expression of integrin alphaV beta3 in mandibular distraction osteogenesis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression and distribution pattern of Integrin alphaV beta3 (ItgalphaV beta3) in course of distraction 44 adults New Zealand white osteogenesis and to quest for the function of ItgalphaV beta3 in distraction osteogenesis (DO).</p><p><b>METHODS</b>rabbits were selected and divided randomly into 3 groups: DO, bone fracture and normal group. There were 20, 20 and 4 rabbits in each group separately. Animal models were established and animals were killed at different time points. Sections were stained by IHC method. Distribution and expression pattern of ItgalphaV beta3 were observed by semi-quantitative technique, and the results were managed with statistic methods.</p><p><b>RESULTS</b>The expression of ItgalphaV beta3 was found both in the DO and bone fracture groups. The positive staining was seen mainly in vascular endothelial cells on cytomembrane and in cytoplasm. The staining intensity of group of DO was higher than that of the bone fracture group.</p><p><b>CONCLUSION</b>ItgalphaV beta3 plays an important role in promoting the process of DO.</p>

The alteration of MTS1 gene in precancerous lesions and squamous cell carcinoma of oral mucosa / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the expression and alteration (including homozygous deletion and mutation) of MTS1 gene in precancerous lesions and squamous cell carcinomas (SCC) of oral mucosa, and to analyse the function of MTS1 gene alteration in oral mucosal carcinogenesis.</p><p><b>METHODS</b>The expression of p16 protein produced by MTS1 gene was examined with immunohistochemical SP method in 10 normal oral mucosas, 30 precancerous lesions (10 mild, 10 moderate and 10 severe dysplasia respectively) and 45 squamous cell carcinomas (SCCI18, SCCII 19, SCCIII 8). The deletion and mutation of exon1 and exon2 of MTS1 gene were examined with methods of PCR and SSCP in these same samples.</p><p><b>RESULTS</b>All the precancerous lesions had p16 protein expression and no alteration of MTS1 gene. In SCC, the positive rate of p16 protein was 60.0% with 72.2% in SCCI, 57.9% in SCCII, 37.5% in SCC III, and there were no significant difference among the three groups by chi2 test (P>0.05). Gene homozygous deletion of exon1 and/or exon2 was detected in 10 cases, and gene mutation in 4 cases. The whole rate of gene alteration was 31.1% (14/45). The MTS1 gene alteration rate was 27.8% in SCCI, 31.6% in SCCII, 37.5% in SCC III and there was also no significant difference among the three groups by chi2 test (P>0.05). In SCC with local lymph nodes metastasis, MTS1 alteration rate was 57.1%, while in SCC with no lymph nodes metastasis was 8.3%, and there was significant difference by chi2 test (P<0.05).</p><p><b>CONCLUSIONS</b>MTS1 gene alteration is not an early event in the carcinogenesis of oral mucosa and can not be used as a biology mark to examine oral precancerous lesions. MTS1 gene may play a certain role in the progression of oral squamous cell carcinomas.</p>