Knockdown of TRIM27 expression regulates cell proliferation, invasion and migration in human nasopharyngeal 5-8F carcinoma cells / 中国病理生理杂志

AIM:To investigate the expression characteristics of TRIM 27 in human nasopharyngeal carcinoma tissues, nasopharyngeal carcinoma 5-8F cells and NP69 cells, and to observe the effect of TRIM27 on the proliferation, in-vasion and migration of 5-8F cells.METHODS:The levels of TRIM27 in the nasopharyngeal carcinoma tissues and normal nasopharyngeal epithelial tissues were observed by the method of immunohistochemistry .The mRNA and protein levels of TRIM27 in the 5-8F cells and NP69 cells were determined by real-time PCR and Western blot .TRIM27 siRNA was trans-fected into the 5-8F cells with Lipofectamine 2000.The relative mRNA expression of TRIM27 was detected by real-time PCR.The relative protein expression of TRIM 27 was detected by Western blot .The cell proliferation was analyzed by CCK-8 assay and cell colony formation assay .The change of cell invasion was examined by Matrigel invasion assay .The change of cell migration were examined by wound healing assay .RESULTS:The results of immunohistochemistry showed that the protein expression of TRIM27 in the nasopharyngeal carcinoma tissues was obviously higher than that in the normal nasopha -ryngeal epithelial tissues .The results of real-time PCR and Western blot showed that the mRNA and protein levels of TRIM27 in the 5-8F cells were obviously higher than those in the NP69 cells.The abilities of proliferation, invasion and migration in the 5-8F cells were significantly suppressed after TRIM27 gene silencing ( P <0.05).CONCLUSION:TRIM27 acts as a oncogene in the 5-8F nasopharygeal carcinoma cells .The abilities of proliferation , invasion and migration are significantly suppressed after TRIM27 gene silencing in the 5-8F cells.

The microsurgical anatomic research of the internal auditory canal area on the retrosigmold approach / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To evaluate the safety of the circular round window and discus anatomic landmarks of posterior wall of internal auditory canal by investigating the microscopic anatomy of internal auditory canal area of the retrosigmold approach, which can provide the anatomical basis for acoustic neutrinomas surgery.@*METHOD@#Fifteen adult cadaver heads (30 sides) fixed with formalin were used in the study. The retrosigmold approach operations were imitated to dissect the blood vessels and nerves in internal auditory canal area by opening round bony window and removing posterior wall of internal auditory canal.@*RESULT@#Fifteen specimens of 30 sides circular bone window were opened without injury with transverse sinus and sigmoid sinus. The vertical distance between the highest point of bone window margo superior and the lowest point of transverse sinus margo inferior was (4.02 ± 0.32) mm. The vertical distance from the most anterior point of bone window leading edge to the most posterior point of sigmoid sinus trailing edge was (6.31 ± 0.43) mm. The internal auditory canal tubercle located in the anterior superior position of internal auditory canal. The vertical distance from the highest point of internal auditory canal tubercle to the upper margin of internal auditory canal was (2.31 ± 0.32) mm. To expose the whole internal auditory canal, the length and width of the internal auditory canal posterior wall removal was (7.29 ± 0.32) mm, (4.12 ± 0.29) mm. Within this removal range, no case of cochlea, semicircular canal or venous was injured in 30 specimens.@*CONCLUSION@#The method of opening round window through retrosigmold approach is simple, practial and convenient. With little variation and easiness of location, the sinternal auditory canal tubercle can be used in the identification of the internal auditory canal. When exposing the whole internal auditory canal, the removal scope of the posterior wall should be paid more attention to, in order to avoid the damage of cochlea, semicircular canal and jugular bulb.

Inhibitory effect of full-length spleen tyrosine kinase on invasion and metastasis of human laryngeal squamous cells / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the effect of gene transfection of full length spleen tyrosine kinase (Syk (L)) on the biological behavior of malignant cancer cells.</p><p><b>METHODS</b>Eukaryotic expression vector pIRES2-EGFP-Syk (L) was constrauted and sequenc. Laryngeal carcinoma cell line Hep-2 were transfected with pIRES2-EGFP-Syk (L) vectors or blank vectors. The expressions of mRNA and protein were examined by real time fluorescence quantitative polymerase chain reaction (Q-RT-PCR) and Western blot analysis. CCK-8 method was used for evaluating cell proliferation, Transwell for cell invasion capacity in vitro, and tumor formation in nude mice for in vivo tumorigenicity.</p><p><b>RESULTS</b>pIRES2-EGFP-Syk (L) vectors were successfully construct and transfected to Hep-2 cells. Q-PCR showed that mRNA expression level in Hep-2 cells transfected with Sky (L) (28.395 ± 0.067) was higher than those in Hep-2-neo cells transfected with blank vectors (3.891 ± 0.021) and Hep-2 cells with no transfection (1.005 ± 0.012), with statistically significant difference (F = 104.02, P < 0.01). Western blot showed that protein expression level of transfected-Sky (L) cells (0.821 ± 0.047) was significantly higher than those of Hep-2-neo cells (0.558 ± 0.031) and Hep-2 cells (0.468 ± 0.031), and the difference was statistically significant (F = 112.32, P < 0.01) ; CCK-8 assay showed OD value (1.390 ± 0.067) of transfected-Sky (L) cells was lower than those of Hep-2-neo cells (1.830 ± 0.067) and Hep-2 cells (1.920 ± 0.040), and the difference was statistically significant (F = 107.64, P < 0.01). Transwell assay showed average cell number per field of transfected-Sky (L) cells (176.04 ± 22.32) was higher than those of Hep-2-neo cells (301.02 ± 21.45) and Hep-2 cells (336.04 ± 26.01) with statistically significant difference (F = 123.46, P < 0.01). The volume (250.77 ± 34.83) mm(3) tumor formed from transfected-Sky (L) cells in nude mice, was less than those from Hep-2-neo cells (750.77 ± 40.83) mm(3) and Hep-2 cells (770.77 ± 30.83) mm(3), with statistically significant difference (F = 165.78, P < 0.01).</p><p><b>CONCLUSION</b>Down-regulation of Syk in Hep-2 cells is associated with the malignant biological behaviors of the cells. Syk (L) may be a potential target in gene therapy for laryngeal squamous cell carcinoma.</p>

miRNA-7 inhibits proliferation of human nasopharyngeal 5-8F carcinoma cells via EGFR/PI3K/Akt pathway / 中国病理生理杂志

AIM: To investigate the relationship of microRNA-7 ( miRNA-7 ) over-expression and epidermal growth factor receptor (EGFR)/phosphatidylinositol kinase-3 (PI3K)/protein kinase B (PKB, also called Akt) pathway in human nasopharyngeal carcinoma 5-8F cells.METHODS:The 5-8F cells were transfected with miRNA-7 mimics (car-rying by Lipofectamine 2000).The expression of miRNA-7 was detected by real-time PCR.The cell proliferation was ana-lyzed by CCK-8 assay.The cell colony-forming capability was determined by cell colony formation test.The expression of EGFR/PI3K/Akt at mRNA and protein levels was examined by real-time PCR and Western blotting.RESULTS:The ex-pression level of miRNA-7 was significantly increased in 5-8F cells compared with negative control ( NC) group and control group ( P<0.01) .The proliferation of NPC 5-8F cells was decreased extremely after tansfected with the miRNA-7 mimics (P<0.01), so did the result of the cell colony-formation test.The expression of EGFR/PI3K/Akt at mRNA and protein levels was significantly down-regulated compared with NC group and control group (P<0.01).CONCLUSION:Over-ex-pression of miRNA-7 significantly inhibits the proliferation and colony-forming ability of nasopharyngeal carcinoma 5-8F cells by down-regulation of EGFR/PI3K/Akt pathway.