| Background: Salivary adenoid cystic carcinoma (SACC), also called cylindroma, isone of the most malignant tumors among gland salivary epithelial malignant tumors.One of the remarkable biological characteristics of this tumor is its early invasion.Because the nerves are often involved, the patients are suffered with pain and nervepalsy, and the corresponding dysfunction. Surgical resection is the preferred method oftreatment, and radical surgical resection need to be expanded. While the postoperativerecurrence rate is still very high and prognosis is very poor, these bring great sufferingto the patients, so better means of treatment is necessitated. There are many factors forthe development of tumors, tumor suppressor genes played very important roles in theprocess, and they become important candidate targets for cancer therapy. At present,many tumor suppressor genes and oncogenes that are closely related with adenoid cysticcarcinoma have been found, such as P53, c-erbB-2, Beclinl and so on. The full name ofPTEN is phosphatase and tensin homologue deleted chromatosome ten. As the firstlyfound tumor suppressor gene which has the phosphorylase function so far, PTEN hasbeen widely concerned in recent years. Studys show that PTEN play an important rolein proliferation, differentiation, apoptosis, adhesion, migration, invasion and many otherbiological behaviors. Loss of PTEN is closely related with tumors, but the role of PTENgene in the development process of the gland salivary adenoid cystic carcinoma hasbeen seldomly reported. SACC-83cell line was isolated from the sublingual glandadenoid cystic carcinoma of a26-year-old woman in1983. Now the cell line has beenwidely used in the development of tumorigenesis and metastasis. For example, the C-erbB2gene was downregulated with RNAi technique in SACC-83cell line, and theproliferation of the SACC-83cell was found to be inhibited. But the role of PTEN genein SACC-83cell line has not been reported.Aim: To study the change of the biological behaviors in salivary adenoid cysticcarcinoma83(SACC-83) cell line and to evaluate the effect of PTEN in thedevelopment of SACC-83by silencing PTEN through RNAi.Method: PTEN-siRNA plasmid was transtected into SACC cell lines (SACC-83)by Lipofectamine2000to knock down the expression of PTEN. Western blot was usedto verify the effect of down-regulation of PTEN at the protein level. The proliferation ofPTEN-siRNA cells were tested by colony formation and CCK8assay. Brduincorporation assay also proofed the effect for cell proliferation after siRNA. Cellinvasion and metastasis ability were evaluated by transwell chamber analysis with orwithout matrigel. Cell metastasis ability also was evaluated by wound healing assay.Cell adhersion ability was evaluated by cell adhersion assay. At last, cell cycle wasevaluated by flow cytometry. Data were analyzed using SPSS17.0for windows.Results:1. PTEN-siRNA was successfully transfected into SACC-83cells and theexpression of PTEN protein expression was efficiently inhibited.2. CCK8assay, Cellcloning experiments and Brdu incorporation assay showed that the cells grow fasterafter PTEN down regulation in vitro.3. Transwell chamber analysis showed that thenumber of PTEN down regulation cells dramatically increased. At the same timewithout matrigel experiments and wound healing assay showed that PTEN downregulation cells movement ability enhancement.4. Cell Adhesion Assay showed that themore ability of the PTEN down regulation cells.5. At last, flow cytometry showed thatthe percent of PTEN down regulation cells in S phase increased and the percent of thecells in phase G1decreased after down-regulation of PTEN.Conclusions: The down regulation of PTEN gene dramatically promote theproliferation, migration and invasion and adhesion of SACC-83cell line. The results ofthese experiments indicate that PTEN gene may play an important role in theoccurrence,development and metastasis of salivary adenoid cystic carcinoma, and could be a therapeutic target in clinical therapy. |
PDF Full Text Request