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Effects of FZD2and FZD3 Genes on the Proliferation and Invasiveness of Salivary Adenoid Cystic Carcinoma Cell

Author: ZhengZuoZuo
Tutor: LuYouGuang
School: Fujian Medical
Course: Clinical Stomatology
Keywords: Adenoid cystic carcinoma Wnt signaling pathway FZD2 FZD3 Cell proliferation Invasive ability
CLC: R739.8
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
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Abstract


Adenoid cystic carcinoma is one of the most common parotid malignancy caused by its unique way of invasion and metastasis of salivary adenoid cystic carcinoma to recur after treatment, is still a lack of effective control measures. Looking from the molecular level to participate in adenoid cystic carcinoma invasion and metastasis-associated molecules, provided a new basis for the diagnosis of adenoid cystic carcinoma, treatment and prognosis. The purpose of the research group pre-confirmed FZD2 occurred FZD3 gene with SACC metastasis, and may play a tumor suppressor role based on further explore FZD2, FZD3 gene expression of adenoid cystic carcinoma cell proliferation and invasion . Methods part for FZD2, gene sequences FZD3 designed and synthesized siRNA transfected SACC-M cells were divided into four groups: siRNA-FZD2 group, siRNA-FZD3 group, NC negative control group and blank group. siRNA transfection using Western blot detection FZD2, the protein expression FZD3, SACC-M cells proliferation using the CCK-8 kit observation applications invasion chamber cell invasion characteristics. Second part the Ad-EasyTM system, the so FZD2, FZD3 genes in E. coli homologous recombination construct Ad-FZD2 Ad-FZD3 adenovirus vectors, restriction endonuclease correct liposomal transfection in AD293 cells in packaging, viral titer was determined the optimal intensity of infection infection SACC-M cells. Using Western blot detection of infection after FZD2, protein's expression FZD3, SACC-M cells proliferation using the CCK-8 kit observed application invasion chamber cell invasion characteristics. The results of the first part of siRNA-FZD2 after 48h, the siRNA-FZD3 transfected SACC-M cells the Western blot display, compared with the negative control group siRNA-FZD2, siRNA-FZD3 histone expression was significantly weakened. SACC-M cells after siRNA interference proliferation curve is located on top of the control group SACC-M cell proliferation curve observed that the application of CCK-8 method SACC-M cells proliferation was markedly increased. The invasion chambers results show after siRNA interference SACC-M cells than the control group SACC-M cells increased significantly through the small number of rooms, the ability of SACC-M cell invasion. The second part of the recombinant adenovirus MOI = 100 infection SACC-M cells after 72h protein immunoblotting, compared with GFP-negative control group the Ad-FZD2 the Ad-FZD3 protein expression was significantly enhanced. Observed that the application of CCK-8 method, SACC-M cells after adenovirus infection growth curves in the control group SACC-M proliferation curve under SACC-M significantly reduced proliferative capacity. The invasion chamber results showed that overexpression FZD2, FZD3 SACC-M cells than the control group SACC-M cells significantly reduced through the small number of rooms, reduced ability of SACC-M cell invasion. Results siRNA interference results. Conclusion FZD2 FZD3 gene in adenoid cystic carcinoma proliferation and invasion play a key role, may be the inhibition of salivary adenoid cystic carcinoma. Further research, looking for new targets to provide the basis for adenoid cystic carcinoma prognosis judgment and gene therapy.

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