Anti-tumor peptide of tumstatin in the treatment of the transplantable model of human laryngeal squamous carcinoma in nude mice / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study the effect of anti-tumor peptide of tumstatin on tumor growth of human laryngeal squamous carcinoma in nude mice and the underlying mechanism.@*METHOD@#Nude mice model bearing laryngocarcinoma were established by using human laryngeal squamous carcinoma cell line (Hep-II). The animals were given tumstatin or PBS for 10 consecutive days. The volumes of the subcutaneous tumor were observed. The microstructure in which the general 2-step immunohistochemical examination was adopted and ultra-micro-structural changes of carcinoma after administration of tumstatin were observed under light and electron microscopes for pathology examination.@*RESULT@#The differences was statistically significant in the net mice weight, tumor weight, tumor volume and tumor weight/net mice weight between the treatment group and the control group (P<0.01). The restrained percentage of tumor was 51.58%. The necrosis and apoptosis of the tumor cells and the angiogenesis reduction were found under light and electron microscope in the treatment group. MVD of the treatment group was lower than that of the control group (P<0.01).@*CONCLUSION@#Tumstatin can significantly restrain the development of laryngocarcinoma.

Apoptosis inducing effect by TRAIL on human laryngeal squamous carcinoma Hep-2 cells / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To study the apoptosis inducing effect of tumor necrosis factor related apoptosis-ligand (TRAIL) on human laryngeal squamous carcinoma Hep-2 cells and its effect mechanism.@*METHOD@#The human laryngeal squamous carcinoma Hep-2 cell line was treated with different concentration of TRAIL in vitro. The inhibition ratio of tumor cells was determined by MTT colorimetric assay, the incidence of cell apoptosis was determined by flow cytometry method. The morphologic changes of laryngeal squamous carcinoma Hep-2 cell were observed with transmission electron microscope.@*RESULT@#In vitro, all the different concentrations of TRAIL inhibited laryngeal squamous carcinoma cell's growth. The inhibited growth ratio showed significant concentration-dependence. The concentrations for inducing apoptosis-ratio(TRAIL 1, 10, 100 microg/L) determined by flow cytometry was (11.49 +/- 0.36)%, (22.31 +/- 0. 82)%, (59.64 +/- 1.10)% respectively in the study group, and (3.13 +/- 0.12)% in the control group, which was significantly different between these two groups (P < 0. 01).@*CONCLUSION@#In vitro, TRAIL inhibited the growth of human laryngeal squamous carcinoma Hep-2 cells. The induced apoptosis of TRAIL shows significant concentration- independence. TRAIL inhibits the growth of human laryngeal squamous carcinoma Hep-2 cells trough inducing apoptosis.

The experimental investigation of laryngeal cancer growth in vive by recombinant lentivirus of small interference RNA (siRNA) targeting MMP-9 / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the inhibitive role of lentivirus mediated MMP-9 gene silence in the growth of laryngeal cancer xenografts.@*METHOD@#The nude mouse model of laryngeal squamous carcinoma was established and RNA interference (RNAi) technic was used. Recombinant MMP-9-RNAi-Lentivirus was intratumoral injected while the empty vector lentivirus was taken as control. After the treatment, tumor inhibitive effect was observed and western-blot was used to test the expression of MMP-9 in the xenografts. Finally, the PCNA expression in xenografts were examined with immunohistochemistry to evaluate proliferation change of the Hep-2 cells.@*RESULT@#The average tumor weight was (1.484 +/- 0.391) g in group treated with MMP-9-RNAi-Lentivirus which was significantly lower than that (2.618 +/- 0.465) g in the control group (P < 0.05). The average tumor volume was (1.177 +/- 0.270) cm3 in group treated with MMP-9-RNAi-Lentivirus which was significantly lower than that (2.034 +/- 0.366) cm3 in the control group (P < 0.05) and the tumor inhibitive rate was 43.32%. Western-blot showed there were 7 cases of xenografts with MMP-9 protein negative expression and the other 3 cases were tested with weak positive expression in the treated group xenografts. Whereas all the 10 cases of xenografts were tested with MMP-9 positive expression in the control group. Immunohistochemistry showed PCNA index in treated group was (55.41 +/- 8.77)% which was obviously lower than that (77.04 +/- 6.91)% in control group (P < 0.01).@*CONCLUSION@#The growth and proliferation of laryngeal cancer could be inhibited by recombinant MMP-9-RNAi-Lentivirus.

The inhibition of laryngeal squamaous cell carcinoma by endostatin / 中国耳鼻咽喉头颈外科

OBJECTIVE To evaluate the inhibitory effect and possibility inhibitory mechanisms ofendostatin on laryngeal carcinoma cells (Hep-2) and human umbilical vein endothelial cells (HUVEC). METHODS ①Using the MTT assay , the effect of different concentrations of endostatin (10 ?g/ml～50 ?g/ml) and endostatin (30 ?g/ml) with different contact times (24～72h) on the livability of Hep-2 and HUVEC cells were determined; ②Changes in the ultrastructures of Hep-2 and HUVEC cells were examined by electron microscopy; ③Survivin mRNA content was determined in Hep-2 cells exposed to endostatin (30 ?g/ml) by RT-PCR test; ④The effect of endostatin (30 ?g/ml) on the ectogenetic artificial blood vessel models was observed by light microscopy. RESULTS ① The growth of Hep-2 and HUVEC cells was significantly inhibited (P

Correlation of matrix metalloproteinases-7 with the invasion, metastasis, and prognostic evaluation in laryngeal squamous cancer / 中国组织工程研究

BACKGROUND: Matrix metalloproteinases-7 (MMP-7) plays an important role in the invasion and metastasis of cancer and is related to prognosis of many carcinomas.OBJECTIVE: To investigate the significance of MMP-7 expression in the invasion and metastasis of laryngeal cancer and explore the relationship between MMP-7 expression and the prognosis of laryngeal squamous cell cancer (LSCC).DEGIGN: An open experimental study based on the cells.SETTING: Two otolaryngology departments of two university hospitals and one otolaryngology department of a municipal hospital.MATERIALS: The experiment was conducted in the Center Laboratory of Second Affiliated Hospital of Harbin Medical University from April 2002 to January 2003. The materials were laryngeal cancer and corresponding neighboring noncancerous tissues, MMP-7 antibody and immunohistochemical kit.METHODS: The expression of MMP-7 protein in 70 samples of paraffin-embedded LSCC and corresponding neighboring noncancerous tissues was detected with immunohistochemical technique. Reverse transcription-polymerase chain reaction(RT-PCR) and Western blot technique were used to examine the expression of MMP-7 in 35 frozen specimens of LSCC and corresponding neighboring noncancerous tissues.MAIN OUTCOME MEASURES: The relationship of MMP-7 mRNA expression with the invasion depth(T grade), neck nodal metastasis and prognosis of LSCC.RESULTS: Immunohistochemical staining of MMP-7 was observed in 77.1% (54/70) of cancer tissues and in 5.7% (4/70) of neighboring noncancerous tissues( P ＜ 0.01) . The mRNA expression of MMP-7 was detected in 74.3% (24/35) of cancer tissues and in 5.7% (2/35) of neighboring noncancerous tissues( P ＜ 0.01 ). The expression of MMP-7 was higher in T3-T4 group than that in T1-T2 group(P ＜ 0.01), and was positively correlated with nodal metastasis( P ＜ 0. 01 ). Western blot showed that both 28 ku (latent form of MMP-7) and 19 ku(active form of MMP-7)bands had close relationship with T grade and neck nodal metastasis. Kaplan-Meier survival curve showed that the group with high MMP-7protein expression had poorer prognosis than the group with weak or negative MMP-7 protein expression(Log-rank = 4. 755 9, 8. 951 3; P = 0. 029 2,0.002 8).CONCLUSION: MMP-7 is closely correlated with the invasion, metastasis,and prognosis of LSCC, and it may serve as a marker in estimating the invasive and metastatic potency and prognosis of LSCC.