Cyr61/CCN1 overexpression induces epithelial-mesenchymal transition leading to laryngeal tumor invasion and metastasis and poor prognosis.

BACKGROUND: To examine the expression of cysteine-rich 61 (Cyr61/CCN1) protein in laryngeal squamous- cell carcinoma (LSCC) tissues, and its relationship with the tumor epithelial-mesenchymal transition (EMT), invasion, metastasis, and prognosis. MATERIALS AND METHODS: Immunohistochemistry was used to detect the expressions of Cyr61, Vimentin (Vim), and E-cadherin (E-cad) in 88 cases of LSCC tissues and 30 cases of tumor-adjacent normal tissues. Vim and E-cad were used as mesenchymal and epithelial markers, respectively, to determine the relationship between Cyr61 expression and the EMT of LSCC cells. In addition, clinical and histopathological data were combined to analyze the relationship between the positive-expression rates of Cyr61, Vim and E-cad and LSCC invasion, metastasis and prognosis. RESULTS: In LSCC tissues, Vim expression rate was significantly higher than that of the tumor-adjacent tissues, whereas E-cad expression rate was significantly lower than that of the tumor-adjacent tissues. The Vim expression rate was significantly higher in stages T3 and T4 than in stages T1 and T2 LSCC tissues, whereas E-cad expression rate was significantly lower in stages T3 and T4 than in stages T1 and T2 LSCC tissues. Compared to the group without lymph node metastasis, the Vim expression rate was significantly higher and the E-cad expression rate was significantly lower in the group with lymph node metastasis. The expression rate of Cyr61 was significantly higher in LSCC tissues than in the tumor-adjacent normal tissues. In addition, the Cyr61 expression rate was higher in stages T3 and T4 than in stages T1 and T2 LSCC, and higher in the group with lymph node metastasis than in the group without lymph node metastasis. The Vim expression rate was significantly higher in the Cyr61 positive group than in the Cyr61 negative group, whereas the E-cad expression rate was significantly higher in the Cyr61 negative group than in the Cyr61 positive group. Survival analysis indicated that survival rates of Cyr61 positive, Vim positive and E-cad negative groups were significantly lower than that of Cyr61 negative, Vim negative and E-cad positive groups, respectively. CONCLUSIONS: Cyr61 expression is closely associated with LSCC invasion and lymph node metastasis. Overexpression of Cyr61 may induce EMT and therefore leads to LSCC invasion and metastasis and poor prognosis. Cyr61 may become a new maker for clinical prediction of LSCC invasion and metastasis and a new target for LSCC treatment.

Cilengitide, a small molecule antagonist, targeted to integrin αν inhibits proliferation and induces apoptosis of laryngeal cancer cells in vitro.

Cilengitide is a chemical synthesis cyclopeptide containing RGD sequence, which can be used as a small molecule antagonist targeted to integrin αν (ITGAV). The aim of present study was to investigate the effect on proliferation and cell apoptosis of the cilengitide in laryngeal cancer cells. In the study, we have treatmented the cultured cells of laryngeal cancer (Hep-2) with cilengitide. After the medication, the proliferation of the Hep-2 cells was detected by MTT assay, the expression of ITGAV was detected by RT-PCR and the activity of caspase-3 protein was detected by a specialized kit. RGD linear peptides (GRGDSP), non-RGD linear peptide (GRGESP), and 5-fluorouracil (5-Fu) were used as controls. Results showed that the proliferation of Hep-2 cells was signally inhibited by the cilengitide with a time and dose compliance. Its inhibition effect was significantly higher than that of 5-Fu and GRGDSP, but the GRGESP showed no obvious inhibitory effect. After intervene of cilengitide, the activity of caspase-3 protein of Hep-2 cells was significantly increased, and the expression of ITGAV was significantly decreased. 5-Fu significantly inhibited the proliferation of Hep-2 cells, but no significant changes of ITGAV expression were observed. In conclusion, cilengitide can significantly down-regulate ITGAV expression and inhibit cell proliferation in laryngeal cancer cells, it will also to induce cell apoptosis through caspase-3 pathway. Therefore, it could be as a kind of effective chemotherapy drugs that will be used in clinical treatment of the laryngeal cancer.

[Transoral CO2 laser epiglottectomy for early epiglottic carcinomas].

OBJECTIVE: To compare the functional results of transoral CO2 laser epiglottectomy with transcervical supraglottic laryngectomy and to find the landmarks for CO2 laser surgery. METHODS: Forty-nine preoperatively untreated patients diagnosed as clinical T1-2N0M0 epiglottic carcinomas enrolled from June 1, 2006, to November 1, 2009 in the Second Affiliated Hospital of Harbin Medical University were studied retrospectively. Seventeen cases were treated by CO2 laser whereas 32 by open surgeries. The upper edge of thyroid cartilage and mucosal recess infra epiglottic tubercle could be used as intraoperation landmarks for transoral CO2 laser surgery. Optional neck dissections (II, III) were performed for 34 patients with tumors on laryngeal face, but not for 15 with tumors on the edges of epiglottis or small laryngeal face tumors. RESULTS: The follow-up for this study covers a period ranging from 3 to 6 years postoperatively. Four local recurrences were found for this study, 2 for laser surgery who underwent additional repeated exisions and 2 for conventional techniques, one received radiotherapy (60 Gy) and another with total laryngectomy. The incidence of lymph node metastasis was 10.2% (5/49) in all cases. Time of naso-feeding or hospitalisation was significantly shorter for CO2 laser treated patients than that for open techniques. Overall 3 year's survival rates were 100% and 90.6% for CO2 laser and the open techniques respectively. CONCLUSIONS: Compared to the open technique, transoral CO2 laser epiglottectomy is a well-tolerated and promising resection technique with low morbidity for early epiglottic carcinomas, and the identification of landmarks is useful for entire resection of epiglottis.

KAI1/CD82 and MRP1/CD9 serve as markers of infiltration, metastasis, and prognosis in laryngeal squamous cell carcinomas.

OBJECTIVE: The current study explored the expression of KAI1/CD82 and MRP1/CD9 and its significance in laryngeal squamous cell carcinoma (LSCC). METHODS: The expression levels of KAI1/CD82 and MRP1/CD9 in 100 LSCC tissue specimens, as well as in 30 para-LSCC non-carcinomatous tissue specimens randomly taken from the patients, were assessed using the quantitative polymerase chain reaction (Q-PCR) and immunohistochemistry and correlations with pathological parameters of LSCC and their influence on survival function were analyzed. RESULTS: KAI1/CD82 and MRP1/CD9 showed basically consistent changes in both mRNA and protein expression. Their expression in the 30 LSCC specimens was significantly lower compared with that in the corresponding non-carcinous tissues (P < 0.01 or 0.05), notably correlating with TNM stage, differentiation degree, clinical stage, and lymphatic metastasis (P < 0.01 or 0.05), but not gender, age, and LSCC growth sites (P > 0.05). The median survival of patients with positive KAI1/CD82 and MRP1/CD9 protein expression was longer than that of patients with negative protein expression (P < 0.01 or 0.05). KAI1/CD82 protein expression negatively correlated with MRP1/CD9 protein expression in LSCC (χ(2) = 31.25, P < 0.01). CONCLUSION: KAI1/CD82 and MRP1/CD9 may jointly participate in the development of LSCC. They may serve as the markers for judging the infiltration, metastasis, and prognosis of LSCC.

Elevated expression of integrin αv and ß5 subunit in laryngeal squamous-cell carcinoma associated with lymphatic metastasis and angiogenesis.

In the past several years, the αv integrin subfamily has been repeatedly found to be involved in tumor progression and angiogenesis. The aim of this study was to investigate the expression of the integrin αv subfamily in laryngeal squamous cell carcinoma (LSCC), and to correlate the expression rate with tumor biological behavior and angiogenesis of the LSCC. The integrin αv subfamily, including αv, ß1, ß3, ß5, ß6 and ß8 subunits, was immunohistochemically found to be expressed in 64 patients with LSCC, and we analyzed the relationship between the expression rate and the clinicopathological stage of this cancer. Immunohistochemical staining for CD105 was carried out in the same group of the patients. The intratumoral microvessel density (IMVD) of the LSCC was calculated by CD105 staining, and the correlation between the IMVD and αv subfamily expression was discussed. The results showed that all members of the integrin αv subfamily could be detected in the LSCC. The expression rate of integrin αv and ß5 subunits in primary cancer was significantly higher than in normal tissue, and their expression rate in the group with lymphatic metastasis was significantly higher than in the group without metastasis. The IMVD of the group with positive expression of αv and ß5 subunits was significantly higher than in the group with negative expression, but there were no significant effects on the ß1, ß3, ß6 and ß8 subunits in these biological processes. In conclusion, the expressions of integrin αv and ß5 subunits were significantly associated with lymphatic metastasis and angiogenesis of the LSCC. Among the members of integrin αv subfamily, integrin αvß5 might play an important role in invasion and metastases of the LSCC, and it may become a valuable marker for the evolution of the LSCC.

Elevated content of osteopontin in plasma and tumor tissues of patients with laryngeal and hypopharyngeal carcinoma associated with metastasis and prognosis.

The aim of this study was to evaluate the usefulness of osteopontin (OPN) expression level in plasma and tumor tissues of patients with laryngeal and hypopharyngeal squamous-cell carcinoma for predicting metastasis and survival of this tumor. The OPN expression in tumor tissues was detected by immunohistochemical staining in a tissue microarray of laryngeal and hypopharyngeal carcinomas, and the OPN level in plasma was measured by ELISA. The expression levels of OPN in plasma and tumor tissues were associated with clinicopathological features and survival of laryngeal and hypopharyngeal carcinomas. Results showed that the OPN expression quantitation either in tissues or plasma was significantly correlated with differentiation and lymphatic metastasis of the laryngeal and hypopharyngeal carcinoma. Elevated OPN level of plasma and tissues was significantly associated with poor survival. In conclusion, elevated OPN level in plasma and tumor tissues was significantly associated with metastasis and survival of laryngeal and hypopharyngeal carcinomas. Elevated OPN level in plasma and tumor tissues may become a useful indicator of prognosis for laryngeal and hypopharyngeal cancers.

Overexpression of osteopontin and integrin αv in laryngeal and hypopharyngeal carcinomas associated with differentiation and metastasis.

PURPOSE: Osteopontin (OPN) is a secreted phosphoprotein, which functions as a cell attachment protein and cytokine that signals through two cell adhesion molecules, integrin αvß3 and CD44, to regulate cancer growth and metastasis. The aim of this study was to investigate the expression of OPN and integrin αv (ITGAV, main receptor of the OPN) in laryngeal and hypopharyngeal squamous cell carcinoma (LHSCC) and correlate the expression quantity with tumor biological behavior. METHODS: We have made a tissue microarray of LHSCC and detected the OPN and ITGAV expression by immunohistochemistry in the microarray. The expression quantity of OPN and ITGAV was determined by a professional pathological image processing software (Image-Pro plus 5.02), and the clinical significances of the expression quantity in LHSCC were analyzed. RESULTS: The expression quantity of OPN and ITGAV in primary and metastatic carcinomas is significantly higher than in normal tissues. The expression of OPN and ITGAV in the well-differentiated group is significantly lower than in moderately and poorly differentiated group; the expression quantity of OPN and ITGAV in group with lymph node metastasis is significantly higher than in group without lymph node metastasis. CONCLUSIONS: The expression of OPN and ITGAV was significantly influenced to the differentiation and metastasis of the LHSCC. Overexpression of the OPN and ITGAV may have contributed to invasion and metastasis of the LHSCC, and therefore, OPN and ITGAV may have value as a target for chemotherapy in LHSCC treatment.

Role of the alpha v-integrin subunit in cell proliferation, apoptosis and tumor metastasis of laryngeal and hypopharyngeal squamous cell carcinomas: a clinical and in vitro investigation.

The objective of this study was to investigate the role of alpha v-integrin subunit (ITGAV, CD51) in invasion and metastasis of the laryngeal and hypopharyngeal squamous cell carcinomas and to evaluate whether an antisense oligonucleotide sequence (ASONs) targeting ITGAV gene can result in proliferative inhibition and induce to apoptosis of laryngeal carcinoma cell lines (Hep-2). Firstly, a tissue microarray contained 75 primary carcinomas, 29 non-cancerous normal tissues and 20 metastatic lymph nodes was constructed and used to detect the expression of ITGAV by immunohistochemistry. The changes of ITGAV expression from each group were assessed and correlated to the clinical parameters of the patients. Secondly, the ASONs targeting ITGAV gene was transfected into Hep-2 cells in vitro. The proliferative ability of the cells after transfection was measured by MTT assay and the apoptosis was detected using flow cytometry. Results showed that the expression of ITGAV was significantly correlated with differentiation and lymph node metastasis of these cancers. In vitro test showed that the proliferative ability of Hep-2 cells was significantly inhibited by ASONs in a way of concentration- and time-depending mode, and a significant apoptosis of Hep-2 cells was also observed after ASONs transfection. In conclusion, the expression of ITGAV was significantly correlated with differentiation and metastasis of the laryngeal and hypopharyngeal carcinomas; down-regulation of ITGAV gene could inhibit proliferation of Hep-2 cells and induce to its apoptosis. These results suggest that ITGAV gene may become a promising prognostic marker and new treatment target for these cancers.

[Expression of alpha-v integrin subunit in the laryngeal and hypopharyngeal squamous cell carcinomas and its relationship with tumor angiogenesis].

OBJECTIVE: To investigate the expression of integrin alpha-v subunit in laryngeal and hypopharyngeal squamous cell carcinomas (LHSCC) and to correlate the expression ratio with clinic and pathologic features of LHSCC, meanwhile, to investigate the relationship between the expression of integrin alpha-v subunit and tumor angiogenesis. METHODS: A tissue microarray of LHSCC was designed and made. Using this microarray, the expression of integrin alpha-v subunit in LHSCC was studied by immunohistochemistry, and the expression disparity in different clinic and pathologic staging of LHSCC was analyzed. The immunohistochemical staining of CD105 was done in the same microarray, the intratumor microvessel density (IMVD) was calculated by CD105 staining. The relationship between integrin alpha-v subunit expression and the IMVD was analyzed. RESULTS: In primary cancer tissue, the expression ratio of integrin alpha-v subunit was 68.0% (51/75), significantly higher than normal tissue beside cancer (10.3%, 3/29, chi2 = 28.68, P < 0.001); the expression ratio of integrin alpha-v subunit in lymph node metastatic carcinoma was 100.00% (20/20), significantly higher than normal tissue (chi2 = 38.77, P < 0.001) and primary cancer tissue (chi2 = 12.69, P < 0.05); in group with lymph node metastasis, the expression ratio of integrin alpha-v subunit was significantly higher than group without lymph node metastasis (chi2 = 10.87, P < 0.001); the IMVD of the group with integrin alpha-v subunit positive expression was significantly higher than the group with integrin alpha-v subunit negative expression (P < 0.001). CONCLUSIONS: There was significant relationship between integrin alpha-v subunit expression and lymphatic metastasis of LHSCC. Overexpression of the integrin alpha-v subunit may have contributed to the tumor angiogenesis and lead to lymphatic metastasis. Integrin alpha-v subunit may become a novel lymphatic metastasis marker of LHSCC.

[Epidemiological study on traffic accident in Shanghai from 2000 to 2002].

OBJECTIVE: Data on traffic accident from year 2000 to 2002 were collected and testified. Epidemiological study was carried out to find the main risk factors of traffic accident. METHODS: The spatial distribution was conducted by means of Geographic Information System (GIS) and were marked on Shanghai digitalized map with different layers by different colors. RESULTS: Results showed that during 2000, 2001 and 2002, 69,669 , 68,894, and 47,088 traffic accident episodes occurred in Shanghai, resulting in 1747, 1724, 1557 deaths respectively with direct economic lost: 2.1, 2.4, 3.0 billion RMB. The main risk factors causing traffic accident deaths would include drinking alcohol before driving, fatigue and speeding. The leading causes of death were head injury and complex injury of body. GIS showed that there was a homocentric circle with more traffic accidents in downtown city, but more deaths in the urban city. Disease burden caused by traffic accident was very heavy. CONCLUSION: In Shanghai, traffic accident has great impact on social and economic issues thus should be controlled effectively. Focus should be layed on control of traffic accident in downtown area and to decrease the number of deaths related to traffic accident in the urban area.

[Laryngeal function reconstruction with sternohyvoid muscle after partial laryngectomy].

OBJECTIVE: To study the laryngeal function reconstruction with sternohyvoid muscle after partial laryngectomy to treat T2 phase of glottis cancer. METHODS: 66 patients diagnosed to be at T2 phase of glottis cancer were treated by partial laryngectomy from 1992 to 1998. At the same time, they were rebuilt vocal cords with sternohyvoid muscle. RESULTS: All patients were decannulated from 2 to 3 weeks. They renewed own respiration, swallow and voice function. Overall three-year survival rate was 97.0% and five-year survival rate was 93.8%. CONCLUSION: New operation is superior than traditional one in improving survival rate and life quality.