Effects of human papillomavirus 16 E6 and E7 oncogene on the proliferation and differentiation of human laryngeal squamous cell carcinoma cell line / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the correlation between HPV16 and the development of laryngeal squamous cell carcinoma by studying the effects of HPV16 E6 and E7 oncogene on the proliferation and differentiation of human laryngeal squamous carcinoma cell line.</p><p><b>METHODS</b>Cationic phosphonolipid was used to transfect pLXSN16E6E7 into Lscc-02 high differentiation laryngeal squamous cell carcinoma cell line, the cells transfected by the vector pLXSN and Lscc-02 cells served as control groups. The changes of proliferation and differentiation were measured in vitro and in vivo.</p><p><b>RESULTS</b>The proliferation was quicker in experimental group. The average A ratio reached to 2.96, which was higher than 1.90 and 1.85 of control groups. The clone forming rate was 23.6% in experimental group, which was higher than 12.7% and 12.0% of control groups. Serum-dependent became lower in experimental group. The Ki-67 positive expression rate was markedly increased and cytokeratin 13 positive expression rate was markedly decreased in experimental group as compared with control groups. The Ki-67 positive expression rates were 93.8%, 80.7% and 79.2% respectively. The cytokeratin 13 positive expression rate were 80.9%, 91.0% and 93.7% respectively. The latent period and internal double time of transplant tumor were short in experimental group compared with control groups. The latent period were 19 d, 28 d and 30 d respectively. The internal double time of transplant tumor was 2.15 d, 3.28 d and 3.47 d respectively. The volume of transplant tumor was smaller in the same period in experimental group. The cells of transplant tumor in experimental group showed an image of small size, great allotype and a trend of low differentiation.</p><p><b>CONCLUSIONS</b>HPV16 E6 and E7 can promote the proliferation and inhibit the differentiation of Lscc-02 laryngeal squamous cell carcinoma cells which suggest HPV16 may play an important role in the development of laryngeal squamous cell carcinoma.</p>

Maxillary swing approach in the management of tumors in the central and lateral cranial base / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>A retrospective review of seventeen patients who were operated through the maxillary swing approach was carried out to assess the efficacy of this approach in the management of tumors of the central and lateral cranial base.</p><p><b>METHODS</b>From May 2000 to January 2005, 17 patients with primary or recurrent neoplasms involving the central cranial or lateral base underwent surgical resection via maxillary swing approach. Ten patients were male, and other seven patients were female, and age range was 7 to 58 years, with a mean age of 42. 6 years. Eight patients had tumors originally involving lateral cranial base, and nine patients had tumors originated from central cranial base. The pathology spectrum was very wide. Among them, five suffered from chordoma, two had rhabdomyosarcoma, two had squamous cell carcinoma, one had malignant fibrous histiocytoma, one had malignant melanoma, one had esthesioneuroblastoma, one had invaded hypophysoma, two had schwannoma, one had pleomorphic adenoma, and one had angiofibroma. Three patients had received previous surgery, two patients had previous radiation therapy and nine patients received postoperative radiotherapy.</p><p><b>RESULTS</b>Sixteen of all seventeen patients had oncologically complete resection, one had near-total resection. This group patients was followed up from 10 to 60 months, with a median follow-up time of 28 months. Two patients died 14 and 26 months after surgery respectively, as a result of local recurrence and metastasis. One patient defaulted follow-up at 12 months after operation, and the other 14 patients were alive at the time of analysis. Of the 12 malignant tumors, the 1-and 2-year survival rate were 91.67% and 72.92%, respectively. The facial wounds of all patients healed primarily, and there were no necrosis of the maxilla, damage of the temporal branch of the facial nerve, lower-lid ectropion, and facial deformity. Epiphora and facial hypoesthesia were detected in all patients. Four patients (23.5%) developed palatal fistula, ten patients developed serous otitis media (58.8%), and four patients developed a certain degree of trismus (23.5%). Cerebrospinal fluid leak occurred in two patients. They subsequently healed with conservative management.</p><p><b>CONCLUSIONS</b>The maxillary swing approach is a proven method for access to the central and lateral skull base with good exposure and acceptable morbidity. Complications and sequelae associated with this approach include facial scarring, transaction of the infraorbital nerve, impaired lacrimal drainage, eustachian tube dysfunction and serous otitis, palatal fistula, trismus etc. Some procedures should be performed for reducing the incidence and severity of complications in the maxillary swing approach.</p>

Differentially expressed gene profiles of cochlea in rats induced by acute and chronic sodium salicylate injection / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the mechanism of electrophysiologic changes caused by different type of sodium salicylate injection.</p><p><b>METHODS</b>Decapitated three group rats ( acute injected, chronic injected and normal rats ) separately, dissected the temporal bones to collect cochlea, and the otic capsules were removed. Then the cochlear materials from each groups were pooled and homogenized respectively, extracted the total RNA, obtained cDNA from purified total RNA by reversed transcription, cDNA were transcripted to cRNA probes in vitro. Hybridized the cRNA probes with tester chip to evaluate the quality of probes, if good, hybridized the probes with real chip. Obtained three gene expression profiles of different groups of cochlea Analyzed the differentially expressed genes among three groups by SOM. Analogized the SOM result to electrophysiologic changes. Then analyzed the genes in clusters of analog results by Gene Ontology. Then the genes in clusters of analog results were analyzed by Gene Ontology. Hsp27 was chosen to validate the result of gene chip using real time quantitative reverse transcription PCR ( RTQ RT-PCR).</p><p><b>RESULTS</b>The probes was good, and the chip hybridization results was credible. We obtained 6 clusters genes by SOM analysis, in which we choose cluster 3 and cluster 4 as candidate cluster. There were 46 genes in cluster 3 and 30 genes in cluster 4 employing GO analysis, which involved in cell communication, cell motility, metabolism, immune response and nerve ensheathment, et al. The result of RTQ RT-PCR showed high concordance with that of gene chip.</p><p><b>CONCLUSION</b>It's a new method to study the mechanism of electrophysiologic changes caused by sodium salicylate by gene chip and SOM analysis.</p>

Establishment of a HPV-negative cell line derived from xenografted human laryngeal squamous cell carcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To establish a human laryngeal carcinoma cell line unassociated with human papillomavirus (HPV).</p><p><b>METHODS</b>Viable tissue of a well-differentiated laryngeal squamous cell carcinoma was obtained and tested negative for the presence of HPV by polymerase chain reaction. Minced tissue fragments were then transplanted subcutaneously into nude mice. After two successive passages, the tumor tissue was seeded into culture flasks and incubated in a medium containing 10% fetal bovine serum, insulin and epidermal growth factor. Tumor cell phenotype and molecular features were determined by various methods.</p><p><b>RESULTS</b>A stable cell line, designated as Lscc-02, was successfully established after 86 culture passages. The cells grew as a monolayer with epithelioid features. The cell doubling time was approximately 39.1 hours. The human origin of the tumor cells was confirmed by karyotype analysis. The squamous epithelial phenotype was demonstrated by the immunopositivity of anti-cytokeratin antibodies and ultrastructural presence of tonofilaments and desmosomes. The malignant nature of the cells was documented by their clonal formation in soft-agar and tumorigenicity in nude mice. Lscc-02 cells expressed carcinoembryonic antigen (CEA) and were negative for HPV DNA.</p><p><b>CONCLUSION</b>This newly established Lscc-02 laryngeal squamous cell carcinoma cell line may be a useful model for investigating laryngeal carcinoma unrelated to HPV infection, and the role of HPV in the progression of human laryngeal carcinoma.</p>