Antitumor effects of Dasatinib on laryngeal squamous cell carcinoma in vivo and in vitro.

A novel drug named Dasatinib is a highly potent ATP-competitive orally active dual Src/Abl kinase inhibitor with anti-proliferative activity against solid tumors and CML (chronic myeloid leukaemia) cell lines. Dasatinib has been shown to have preclinical activity against human prostate, breast, pancreatic, lung, and head and neck cancer. To determine whether Dasatinib can inhibit the growth of laryngeal squamous cell carcinoma, in the present study, we investigated the antitumor effect of Dasatinib on Hep-2 cells. Hep-2 cells were treated with different concentrations of Dasatinib for different time. Cell proliferation, cell cycle distribution, and cell apoptosis were evaluated using MTT assay, flow cytometry, and fluorescent microscopy. It was found that Dasatinib exhibited significant efficacy in growth inhibition, cell cycle arrest at G0/G1 phase, and apoptosis induction in a dose- and time-dependent manner. Measuring the modulation of regulators in the cell cycle, apoptosis and signal transductions by western blot analysis showed that the effect of Dasatinib was due to suppression of the expression of Bax, Bcl-2, Caspase-3, and Caspase-8. Moreover, in vivo studies were performed in a nude mouse xenograft model, the new prescription (DDP + Dasatinib) was better than DDP alone in terms of therapeutic efficacy. In conclusion, the antitumor effect of Dasatinib on Hep-2 cells was due to the induction of cell cycle arrest as well as apoptosis. The possible mechanisms underlying the action might be attributed to the suppression of Src phosphorylation. This investigation suggests a potential clinical application of Dasatinib for the treatment of laryngeal cancer patients.

Antitumor and immunoregulatory effects of astragalus on nasopharyngeal carcinoma in vivo and in vitro.

This study was carried out to evaluate the effects of Astragalus on human nasopharyngeal carcinoma (NPC) viability and apoptosis and to investigate the mechanism of Astragalus in a NPC cell line (CNE2). Cell viability was measured using the MTT assay. CNE2 cells treated with Astragalus were stained with acridine orange/ethidium bromide and subjected to fluorescence microscopy. Bcl-2, Bax, caspase-3 and -8 were measured by western blotting. Rat NPC cells were used to establish a NPC model. Tumor weight, immune organ index and T lymphocyte subsets were employed to detect the immunoregulatory and antitumor effects of Astragalus after administration. Astragalus was effective in inducing apoptosis in CNE2 cells. Morphological changes associated with cell injury were found. Western analysis showed caspase-3, -8, and Bax protein levels were increased after Astragalus treatment, while the bcl-2 protein level was decreased. Astragalus increased the percentage of CD3(+) , CD4(+) T-lymphocytes, and the ratio of CD4(+) /CD8(+) . Astragalus also restored the immunological effects of DDP-induced immunosuppression. These findings suggest that the immunomodulatory and anticancer effects of DDP + Astragalus were better than those of DDP alone, and Astragalus could inhibit immunosuppression induced by DDP. The combination of CDDP + Astragalus could be developed as an effective chemotherapeutic regimen in the treatment of nasopharyngeal carcinoma.

Overexpression of SATB1 in laryngeal squamous cell carcinoma.

OBJECTIVE: To investigate the expression of special AT-rich sequence-binding protein 1 (SATB1), and the relationship between SATB1 and clinicopathological factors of laryngeal squamous cell carcinoma (LSCC). METHODS: Real-time PCR, Western blot and immunohistochemistry were used to analyze 80 samples of LSCC and 25 samples of control mucosa. The relationship between SATB1 expression and clinicopathological parameters was evaluated. RESULTS: The SATB1 mRNA expression levels in LSCC were 2.5- to 7.5-fold higher than those in control mucosa tissues (p < 0.001). SATB1 protein expression was detected in approximately 66% (53 out of 80) of the LSCC specimens, whereas it was below the detection limit in all the control mucosa specimens (p < 0.001). The SATB1 mRNA levels in positive cervical lymph nodes, in clinical stages III and IV with poor/moderate cell differentiation, were significantly higher than those in negative cervical lymph nodes in clinical stage II with high cell differentiation (p = 0.022, p = 0.005 and p = 0.006, respectively). CONCLUSIONS: SATB1 mRNA and protein levels are elevated in LSCC tissues, and their levels are correlated with clinical stages and differentiation status. The current findings suggest that SATB1 may be a useful marker for the prognosis and assessment of therapeutic effects.

MicroRNA-1469, a p53-responsive microRNA promotes Genistein induced apoptosis by targeting Mcl1 in human laryngeal cancer cells.

Genistein, a plant isoflavone, is reported to have therapeutic potentials in multiple cancers, However, the molecular mechanism underlying promoting cell apoptosis in laryngeal cancer remains unclear. In this study, we report that miR-1469 was induced by genistein in laryngeal cancer. Elevated miR-1469 promoted cell apoptosis and inhibited Mcl1 expression. In addition, we also observed that tumor suppressor p53 was increased under genistein treatment. Elevation of p53 promoted miR-1469 expression, leading to miR-1469 increase and Mcl1 decrease. Therefore, our findings suggest that genistein can suppress laryngeal cancer cell survival through p53 -miR-1469-Mcl1pathway.

Missing diagnosis of neck metastases by routine detecting method in laryngeal carcinomas.

OBJECTIVE: To evaluate the missing diagnosis of neck metastases by routine detecting method (palpation combined with one pathological slide) in laryngeal carcinomas. METHODS: Sixty-six specimens of neck dissections were collected and observed by routine method, transparent method, and continuous sliding method. RESULTS: Totally, 1153 lymph nodes were detected by palpation method and another 1204 lymph nodes were detected by transparent method. The lymph nodes detected by transparent method account for 51.1% of the total, and among them 10 metastases were found, which account for 15.6% (10/64) of metastatic lymph nodes. For those with no metastasis detected by routine method, 50 microm interval continuous sliding method was performed, and 14 tiny metastases were found, which account for 21.9% (14/64) of metastatic lymph nodes. Detecting by routine method, most lymph nodes (95%) were in tumor growth and tumor suffusion stage. The missing diagnosis rate of routine method was 37.5% (24/64). CONCLUSIONS: When routine method was used to detect lymph nodes in neck specimens, missing diagnosis should be considered to select best therapy. Through transparent method small lymph nodes could be found and it is a valuable method to observe pathological changes of small nodes. Continuous sliding method could find micrometastasis precisely, but the work burden is heavy and it is difficult to be widely used.

Extracapsular spread in ipsilateral neck metastasis: an important prognostic factor in laryngeal cancer.

OBJECTIVE: To evaluate the impact of extracapsular spread (ECS) in ipsilateral neck metastasis on prognosis and its related factors in laryngeal cancer. METHODS: The study included 184 patients who underwent laryngectomy and simultaneous radical or modified radical neck dissection between January 1994 and December 1997 for laryngeal cancer. All of them had a complete 5-year follow-up. We used transparent lymph node detection and continuous slicing method on all neck dissection specimens. Kaplan-Meier model was used for survival analysis and the log-rank test was used to assess significance. RESULTS: We found pathological neck metastases in 80 patients. Among them, 26 cases (32.5%) had ECS in ipsilateral neck. ECS incidence increased with advanced pathological N (pN) stages (pN1 3.7%, pN2a 25.0%, pN2b 50.0%, and pN2c 55.6%; P = 0.001). ECS incidence also increased with number of positive nodes (1 positive node 8.6%, 2 positive nodes 33.3%, 3 and more positive nodes 66.7%; P < 0.001). Incidences of contralateral neck metastases and ipsilateral neck recurrence in patients with ECS were higher than those in patients without ECS (46.2% vs. 24.1%, P = 0.046; 34.6% vs. 7.4%, P = 0.002). The 5-year survival rate of patients with ECS was significantly lower than that of patients without ECS (23.1% vs. 57.4%, P = 0.013). CONCLUSION: ECS is an important prognostic factor in laryngeal cancer. Patients with ECS have a higher incidence of contralateral neck metastasis, so bilateral neck dissection should be selected.

[Impact of extracapsular lymph node spread in the ipsilateral neck on contralateral neck metastasis and prognosis of laryngeal cancer].

OBJECTIVE: To evaluate the impact of extracapsular lymph node spread (ECS) in the ipsilateral neck on the contralateral neck metastasis and prognosis of laryngeal cancer. METHODS: The data of 184 laryngeal cancer patients who underwent laryngectomy and simultaneous radical or modified radical neck lymph node dissection between Jan. 1994 and Dec. 1997 were retrospectively analyzed. Of these 184 patients, 144 underwent unilateral neck lymph node dissection and 40 bilateral; 159 had supraglottic lesion and 25 transglottic. All had squamous cell carcinoma. The clinical T stage was T1 in 3, T2 63, T3 77, T4 41; N stage: NO in 123, N1 38, N2a 5, N2b 11, N2c 7. Transparent lymph node detection and continuous sectioning method were applied to all dissected neck lymph nodes. Statistical analysis was carried out using SPSS software package ( version 11.5). Survival curves were calculated through the Kaplan-Meier model. Impact of extracapsular lymph node spread in the ipsilateral neck on prognosis was assessed using the Log rank test. RESULTS: Of these 184 patients, neck lymph node metastasis was pathologically proven in 80, 26 had ECS in the ipsilateral neck with a ECS rate of 32.5% (26/80). The ECS incidence was positively correlated with advanced pathological N stage and metastatic lymph nodes (P < 0.01). The incidence of the contralateral neck metastasis and ipsilateral neck recurrence with ECS were higher than those without ECS, which was 46.2% versus 24.1%, and 34.6% versus 7.4%, respectively (P < 0.05). The 3- and 5-year survival rates of patients with ECS were significantly lower than those of patients without ECS, which was 53.9% versus 70.4%, and 23.1% versus 57.4%, respectively (P = 0.0125). CONCLUSION: Extracapsular lymph node spread is found to be an important prognostic factor in the laryngeal cancer. Bilateral neck dissection may be mandatory due to patients with ECS have a higher incidence of contralateral neck metastasis. The capsule of metastatic lymph nodes should be pathologically checked and reported in order to determine the extra-capsular spread status.

[Prognostic factors identified by Cox multivariate analysis of surgically treated 1018 laryngeal cancer patients].

OBJECTIVE: To study the prognostic factors of 1018 patients with laryngeal cancer treated surgically. METHODS: All patients were treated surgically for laryngeal cancer from 1984 to 1996. A total of 16 clinical factors was studied by univariate analysis and Cox multivariate model. RESULTS: The follow-up rate was 93.5% over 5 years. The overall cumulative survival rate was 79.1% at 3 years, 70.2% at 5 years. The 5-year survival rate of T1N0 is the highest, followed by T1N+, T2N0, T3N0, T4N0, T2N+, T4N+, and T3N+. In univariate analysis, the survival was related to patient age, mobility of vocal cords, preoperative T status, preoperative N status, preoperative UICC stage, postoperative T status, postoperative N status, postoperative UICC stage, topographic location of the tumor and tumor size. In Cox multivariate modal, only postoperative N status, mobility of vocal cords and tumor size were independent prognostic factors. CONCLUSION: Independent prognostic factors for patients with laryngeal cancer after curative resection are postoperative N stage, mobility of vocal cords as well as tumor size. Postoperative follow-up and salvage surgery in time should be attached with importance to improve the survival of patients with laryngeal cancer.

MicroRNA-221 accelerates the proliferation of laryngeal cancer cell line Hep-2 by suppressing Apaf-1.

Laryngeal cancer is one of the most commonly occurring malignant cancers of the head and neck region. In the present study, we investigated the roles of miR-221 in laryngeal squamous cell carcinoma cell line, Hep-2. We examined the function and mechanism of miR-221 in Hep-2 cells using techniques of cell biology and molecular pathology, such as western blotting, quantitative PCR, immunohistochemical staining and flow cytometry. Using a luciferase assay, the apoptotic protease activating factor-1 (Apaf-1) mRNA 3'-UTR was shown to have complementary binding sites using bioinformatics prediction software including TargetScan, PicTar and miRanda. In conclusion, our results showed that miR-221 inhibition caused elevated expression levels of the Apaf-1 apoptotic pathway proteins caspase-3, -8 and -9. miR-221 may therefore be used as a novel therapeutic target for laryngeal cancer.

Clinical significance of epigenetic silencing and re-expression of O6-methylguanine-DNA methyltransferase using epigenetic agents in laryngeal carcinoma.

The aim of the present study was to investigate the association between O6-methylguanine-DNA methyltransferase (MGMT) gene expression levels, and DNA methylation status and histone modifications in laryngeal squamous cell carcinoma (LSCC). Chromatin immunoprecipitation, methylation-specific polymerase chain reaction (PCR), and reverse transcription-quantitative PCR were performed to analyze histone modifications, DNA methylation status and mRNA expression levels in the promoter region of the MGMT gene in laryngeal carcinoma HEp-2 cells, as well as in 50 paired healthy and LSCC tissue samples. The present study demonstrated that treatment of HEp-2 cells with 5-aza-2'-deoxycytidine (Aza), a DNA methyltransferase inhibitor, significantly upregulated MGMT mRNA expression levels, reduced MGMT DNA methylation, reduced MGMT histone H3 lysine 9 (H3K9) di-methylation, and increased MGMT histone H3 lysine 4 di-methylation without a significant change in H3K9 acetylation. Trichostatin A (TSA), a histone deacetylase inhibitor, marginally upregulated MGMT mRNA expression levels without affecting the DNA methylation status, or H3K9 or H3K4 di-methylation, however, TSA treatment caused a significant increase in H3K9 acetylation. Furthermore, Aza and TSA combination treatment produced a synergistic effect. In the LSCC samples, the rate of DNA methylation in the MGMT gene was 54%, compared with 24% in the healthy control group (P<0.05). Therefore, data from the present study indicates that MGMT may serve as a novel therapeutic target in the treatment of LSCC.

MicroRNA-126 modulates the tumor microenvironment by targeting calmodulin-regulated spectrin-associated protein 1 (Camsap1).

Plasma miRNAs have been reported as biomarkers for various diseases. In this study, we investigated whether plasma concentrations of miR-126 may be useful as biomarkers for laryngeal squamous cell carcinoma (LSCC). We examined the function and mechanism of miR-126 in LSCC by using cell biology and molecular pathology techniques such as western blotting, quantitative PCR, IHC and IF. The expression of Camsap1 mRNA and protein is higher in cancer tissues compared to that in normal tissues. Both miR-126 and Camsap1 were related with the prognosis of LSCC patients. We found that miR-126 was able to inhibit LSCC partly by suppressing Camsap1 expression. In addition, Camsap1 expression induced microtubule formation and aggregation. This mechanism possibly explains why loss of miR-126 is frequently associated with tumor metastasis.

Overexpression of miR -155 promotes proliferation and invasion of human laryngeal squamous cell carcinoma via targeting SOCS1 and STAT3.

MicroRNA155 plays an important role in many solid malignancies. Expression and function of miR-155 in laryngeal carcinoma have not been fully understood. This study aims to investigate the expression and function of miR-155 in laryngeal squamous cell carcinoma (LSCC), the relationship between miR-155 and its downstream target suppressor of cytokine signaling 1 (SOCS1)-STAT3 pathway, and the related clinicopathological factors. Sixty-three samples of laryngeal squamous cell carcinoma and twenty-one samples of control mucosa obtained from total laryngectomy cases were analyzed using Western blot analysis and real-time PCR. Hep-2 cells were cultured and transfected with miR-155 mimic and ASO. Cell proliferation, migration and invasion assays were used to determine the role of miR-155 in regulation of LSCC growth, migration, and invasion, respectively. The expression levels of miR-155 in LSCC were significantly higher than those in the control mucosa tissues. Downregulation of SOCS1 expression and elevated expression of STAT3 were also observed in LSCC. The relevance of the three factors were statistically significant. Moreover, knockdown of miR-155 elevated SOCS1expression level, suppressed STAT3 expression, and inhibited hep-2 cells growth, migration and invasion. Whereas overexpression of miR-155 inhibited SOCS1expression, elevated STAT3 expression, and promoted hep-2 cells growth, migration and invasion. Furthermore, the miR-155 levels in T(3) T(4) stages, and poor/moderate cell differentiation were significantly higher than those in T(2) stage and higher degree of cell differentiation. The STAT3 protein in poor/moderate cell differentiation was significantly higher than those in higher degree of cell differentiation. We firstly demonstrated the aberrant expression and function of miR-155 and itsdownstream targets in LSCC. The current findings suggest that miR-155 play promotingrole during the development of LSCC, and miR-155 may be a useful marker for the prognosis and assessment of therapeutic effects.

Inhibition of p21-activated kinase 4 expression suppresses the proliferation of Hep-2 laryngeal carcinoma cells via activation of the ATM/Chk1/2/p53 pathway.

In the present study, we investigated the effects of the p21-activated kinase 4 (Pak4) gene on Hep-2 laryngeal carcinoma cells in vivo and in vitro. The expression of Pak4 was downregulated using small interfering RNA (siRNA). The downregulation of Pak4 decreased the proliferation and increased apoptosis and S phase arrest in Hep-2 cells in vitro. In further experiments, we determined that the S/G(2) transition was obstructed by the downregulation of Pak4 using 5­chloro-2'­deoxyuridine (CldU) and 5­iodo­2'­deoxyuridine (IdU) double staining. A xenografted Hep-2 tumor mouse model was created by inducing human tumors with a subcutaneous (s.c.) injection of 5x10(6) Hep-2 cells into the dorsal flank region of nu/nu mice. The downregulation of Pak4 in established xenografted tumors decreased tumor size and weight. The survival rate of the mice with tumors that did not express Pak4 was significantly higher compared to the mice with tumors expressing Pak4. These results confirm the role of Pak4 as an oncogene in laryngeal carcinoma cells. To identify the mechanism of the cell cycle arrest induced by Pak4, immunohistochemical staining was performed to detect changes in cell cycle­related proteins. The results demonstrated that p53 was activated following the downregulation of Pak4. The levels of ataxia telangiectasia mutated (ATM), the upstream protein of checkpoint kinase (Chk)1 and Chk2, also increased. Therefore, we confirmed that the mechanisms of the Pak4-induced cell cycle arrest invovlve the activation of the ATM/Chk1/2/p53 pathway. These results may prove helpful for the development of novel therapies for the treatment of laryngeal carcinoma.

The tobacco-specific carcinogen NNK induces DNA methyltransferase 1 accumulation in laryngeal carcinoma.

Over-expression of DNA methyltransferase 1 (DNMT1) correlates with hypermethylation of tumor suppressor genes (TSGs) in tobacco-induced cancers. The tobacco component nitrosamine 4-(methylnitro-samino)-1-(3-pyridyl)-1-butanone (NNK) increases protein levels of the DNMT1 in human lung cancer. However, the role of DNMT1 expression induced by NNK is not clear during laryngeal carcinogenesis. We investigated DNMT1 expression levels in 101 cases of human laryngeal carcinoma specimens and 54 cases clear surgical margin specimens by reverse transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry (IHC) detection. Then, we analyzed the relationship between the DNMT1 expression and the smoking status of the patients with laryngeal carcinoma. Moreover, we investigated the effects of tobacco carcinogen NNK on DNMT1 expression in Hep-2 cells. We found that DNMT1 mRNA and protein expressions were up-regulated in laryngeal cancer tissues (p<0.05 and p<0.01, respectively). Among the 101 cases, DNMT1 protein from patients with heavier smoking habit had a significant trend of an increase with IHC scores (p<0.01). The overall survival rates of patients DNMT1-positive were significantly lower than those of patients DNMT1-negative (p<0.05). We observed that NNK increased DNMT1 protein levels, not mRNA levels, in cultured Hep-2 cells significantly in both dose- and time-dependent manner (p<0.05). These results supported the idea that NNK-induced DNMT1 expression may result from protein stabilization. Increased DNMT1 protein expression may play a critical role in the malignant progression of larynx.

[DNA methylation and histone modification relate to RASSF1A gene deletion in laryngeal carcinoma tissues].

OBJECTIVE: To investigate the relationship between RASSF1A gene expression and DNA methylation or histone modification in laryngeal carcinoma tissues. METHODS: Chromatin immunoprecipitation (ChIP), methylation specific polymerase chain reaction (MSP) and realtime quantitative reverse transcription polymerase chain reaction (realtime RT-PCR) were used to analyze RASSF1A gene promoter region histone H3 lysine 9 methylation, H3 lysine 4 methylation, H3 lysine 9 acetylation, DNA methylation, and RASSF1A gene expression in laryngeal carcinoma tissue of 50 cases. RESULTS: DNA methylation rate of gene RASSF1A was 62% in 50 cases of laryngeal carcinoma, but no DNA methylation was found in normal control group, with a significant difference (χ(2) = 15.381, P < 0.05). DNA methylation had no correlation with age, gender, differentiation degree, T stage, pathological type and lymph node metastasis (P > 0.05). The affection of DNA methylation group was more than unmethylation group to expression of gene RASSF1A (t = -3.108, P < 0.01). There was positive correlation between RASSF1A deletion and gene hypermethylation or between H3 lysine 9 methylation of RASSF1A gene promoter and DNA methylation in laryngeal carcinoma tissue(r = 0.816, P < 0.05), but there was negative correlation between H3 lysine 4 methylation of RASSF1A gene promoter and DNA methylation (r = -0.837, P < 0.05) and no correlation between H3 lysine 9 acetylation and DNA methylation (r = -0.383, P > 0.05). CONCLUSIONS: Laryngeal tumor suppressor gene RASSF1A promoter methylation is a key factor down-regulating the gene expression, and histone modifications also plays an important role in tumor development.

[Reconstruction of laryngeal defect in vertical partial laryngectomy with resection of arytenoid cartilage].

OBJECTIVE: To discuss the method to reconstruct laryngeal defect after vertical partial laryngectomy with resection of arytenoid cartilage. METHODS: Laryngeal defect was reconstructed with local tissues after vertical partial laryngectomy with resection of arytenoid cartilage on 87 patients with laryngeal carcinoma of glottic type (T1 7 cases, T2 54 cases, T3 26 cases). All the lesions invaded arytenoid area or vocal process. No filling tissues were used to increase the height of affected arytenoid area and no skin flap or other tissues were used to reconstruct the vocal cord in all the patients. RESULTS: All the patients recovered normal swallow in 8 to 19 days postoperation and restored phonation. The decannulation rate was 98.9% (86/87). There were no pharyngeal fistula and pulmonary complications after operation. Local infection occurred in 3 patients and was cured in 7 days. The rate of local recurrence and cervical lymph node metastasis were 8.0% (7/87), 6.9% (6/87) respectively. Lost patients were assumed to death and direct method was used to calculate survival rate. In 87 patients postoperative period was above 3 years, 5 died in 3 years and 3 were lost 3- year survival rate was 90.8% (79/87). In 63 patients postoperative period was above 5 years, 10 died in 5 years and 2 were lost. 5- year survival rate was 81.0% (51/63). CONCLUSIONS: Utilizing local tissues to reconstruct laryngeal defect after vertical partial laryngectomy with resection of arytenoid cartilage will not lead to severe dysphagia. Phonation is acceptable. It not only saves the operation time but also avoids the negative effects of immoderate reparation.

[Preliminary study on the correlation factors of cervical lymphatic metastasis of glottic carcinoma].

OBJECTIVE: To investigate the incidence of occult nodal in glottic carcinoma and access the correlation between the cervical lymph nodes metastases and originals of tumor. METHODS: A retrospective review was made on 452 patients from January 1983 to May 1998. 413 patients were male and 39 were female. The age ranged from 42 to 79 years with a mean of 59.3 years. RESULTS: The incidence of occult nodal in glottic carcinoma was low (3.54%), and 0.29% in early stage (T1-T2), 13.39% in late stage (T3-T4), respectively. The number of metastasis lymph nodes was 7 in level II (43.75%), 7 in level III (43.75%) and 2 in level IV (12.5%). CONCLUSION: According to the multivariate analysis, none of the factors such as histopathologic differentiation and invasion of peripheral tissue significantly affect on cervical metastases. The incidence of cervical lymph nodes metastases in glottic carcinoma is low. Selective neck resection should not be undertaken even for the advanced stage that don't exist metastases.

[Survival analysis of 1115 patients with laryngeal carcinoma].

OBJECTIVE: To study the long-term result of patients with laryngeal carcinoma treated by surgery and the prognostic factors. METHODS: The survival status of 1115 patients with laryngeal carcinoma which were treated in this department between 1983 and 1996 were retrospectively investigated. RESULTS: Overall 5-year survival rate was 77%. Among them, 5-year survival rate for patients of stage I was 94%, stage II 89%, stage III 82%, stage IV 66%. Patients with glottic cancer had the best prognosis, followed by supraglottic, subglottic and transglottic cancer. Five years survival rate for patients with partial laryngectomy was 85%, whereas, for total laryngectomy was 68%. The causes of failure were local recurrence and metastasis(70%). 14% of dead was not clear. CONCLUSION: Early diagnosis was the key for both larynx preservation and survival rates. Causes of dead were laryngeal recurrence and metastasis.

[Expression of the metastasis-associated gene 1 in laryngeal squamous cell carcinoma: correlation with cervical lymph node metastasis].

OBJECTIVE: The purpose of study was to examine the mRNA expression levels of the metastasis-associated gene1 (MTA1) in laryngeal squamous cell carcinoma (LSCC), so to evaluate its relationship with metastases. METHODS: Forty-eight surgically resected primary LSCC (16 supraglotti laryngeal carcinomas with cervical lymph node metastasis, 16 supraglotti laryngeal carcinomas with cervical lymph node-negative metastasis, 16 glottic carcinoma with cervical lymph node-negative metastasis) and 16 normal laryngeal mucosa was examined for mRNA expression of MTA1 by reverse transcription-polymerase chain reaction(RT-PCR). RESULTS: The frequency of MTA1 mRNA positive expression in 16 LSCC with cervical lymph node metastasis was 100%, but no expression of MTA1 mRNA was observed in other LSCC with cervical lymph node-negative metastasis (16 supraglotti laryngeal carcinomas and 16 glottic carcinoma) and 16 normal laryngeal mucosa. CONCLUSION: MTA1 gene is strongly related to cervical lymph node metastasis of LSCC, and may serve as a early diagnosis indicator.