Abstract: Objective To construct eukaryotic expression vectors of RNA interference specific to matrix metalloproteinase 1(MMP1). Methods Genome sequences of MMP1 gene were retrieved from Genbank and cDNA that was capable of coding expression of shRNA (small hairpin RNAs) was designed for MMP1 gene. The cDNA was synthesized and inserted into plasmid pWH1, and the recombinant pWH1-MMP1 expression vector was identified using enzyme cutting. Then, pWH1-MMP1 expression vector was transfected into salivary adenoid cystic carcinoma ACC-M cells. Finally, the expression of MMP1 in ACC-M cells transfected with pWH1-MMP1 expression vector was evaluated. Results Compared with ACC-M cells and ACC-M cells transfected with plasmid pWH1, the ACC-M cells transfected with pWH1-MMP1 expression vector showed a low mRNA and protein expression of MMP1. Conclusions The constructed MMP1 shRNA expression vector can block the expression of MMP1 in salivary adenoid cystic carcinoma cells, potentially facilitating further studies on the significance of MMP1 in the proliferation, metastasis and experimental treatment of salivary adenoid cystic carcinoma.
Jordan R C, Macabeo-Ong M, Shiboski C H, et al. Overexpression of matrix metalloproteinase-1 and -9 mRNA is associated with progression of oral dysplasia to cancer[J]. Clin Cancer Res, 2004,10(19):6460-6465.
[1]
Sunami E, Tsuno N, Osada T, et al. MMP-1 is a prognostic marker for hematogenous metastasis of colorectal cancer[J]. Oncologist, 2000,5(2):108-114.
[2]
Jordan R C, Macabeo-Ong M, Shiboski C H, et al. Overexpression of matrix metalloproteinase-1 and -9 mRNA is associated with progression of oral dysplasia to cancer[J]. Clin Cancer Res, 2004,10(19):6460-6465.
[3]
Toruner G A, Ulger C, Alkan M, et al. Association between gene expression profile and tumor invasion in oral squamous cell carcinoma[J]. Cancer Genet Cytogenet, 2004,154(1):27-35.
[3]
Toruner G A, Ulger C, Alkan M, et al. Association between gene expression profile and tumor invasion in oral squamous cell carcinoma[J]. Cancer Genet Cytogenet, 2004,154(1):27-35.
[4]
Bendardaf R, Lamlum H, Vihinen P, et al. Low collagenase-1 (MMP-1) and MT1-MMP expression levels are favourable survival markers in advanced colorectal carcinoma[J]. Oncology,2003,65(4):337-346.
[5]
Yamashita K, Mori M, Kataoka A, et al.The clinical significance of MMP-1 expression in oesophageal carcinoma[J]. Br J Cancer,2001,84(2):276-282.
[4]
Bendardaf R, Lamlum H, Vihinen P, et al. Low collagenase-1 (MMP-1) and MT1-MMP expression levels are favourable survival markers in advanced colorectal carcinoma[J]. Oncology,2003,65(4):337-346.
[6]
Ito T, Ito M, Shiozawa J, et al. Expression of the MMP-1 in human pancreatic carcinoma: relationship with prognostic factor[J]. Mod Pathol,1999,12(7):669-674.
[5]
Yamashita K, Mori M, Kataoka A, et al.The clinical significance of MMP-1 expression in oesophageal carcinoma[J]. Br J Cancer,2001,84(2):276-282.
[7]
Inoue T, Yashiro M, Nishimura S, et al. Matrix metalloproteinase-1 expression is a prognostic factor for patients with advanced gastric cancer[J]. Int J Mol Med,1999,4(1):73-77.
[6]
Ito T, Ito M, Shiozawa J, et al. Expression of the MMP-1 in human pancreatic carcinoma: relationship with prognostic factor[J]. Mod Pathol,1999,12(7):669-674.
[8]
Boire A, Covic L A, Agarwal A, et al. PAR1 is a matrix metalloprotese-1 receptor that promotes invasion and tumorigenesis of breast cancer[J]. Cell, 2005,120(3):303-313.
[7]
Inoue T, Yashiro M, Nishimura S, et al. Matrix metalloproteinase-1 expression is a prognostic factor for patients with advanced gastric cancer[J]. Int J Mol Med,1999,4(1):73-77.
[9]
Lum L,Beachy P A. The Hedgehog response network: sensors, switches, and routers[J]. Science,2004,304(5678):1755-1759.
[8]
Boire A, Covic L A, Agarwal A, et al. PAR1 is a matrix metalloprotese-1 receptor that promotes invasion and tumorigenesis of breast cancer[J]. Cell, 2005,120(3):303-313.
[9]
Lum L,Beachy P A. The Hedgehog response network: sensors, switches, and routers[J]. Science,2004,304(5678):1755-1759.
[10]
Liu L T, Chang H C, Chiang L C, et al. Histone deacetylase inhibitor up-regulates RECK to inhibit MMP-2 activation and cancer cell invasion[J]. Cancer Res,2003,63(12):3069-3072.
[11]
Lima R T, Martins L M, Guimaraes J E, et al. Specific downregulation of bcl-2 and xIAP by RNAi enhances the effects of chemotherapeutic agents in MCF-7 human breast cancer cells[J]. Cancer Gene Ther,2004,11(5):309-316.
[10]
Liu L T, Chang H C, Chiang L C, et al. Histone deacetylase inhibitor up-regulates RECK to inhibit MMP-2 activation and cancer cell invasion[J]. Cancer Res,2003,63(12):3069-3072.
[12]
Berns K, Hijmans E M, Mullenders J, et al. A large-scale RNAi screen in human cells identifies new components of the p53 pathway [J]. Nature, 2004, 428(6981):431-437.
[11]
Lima R T, Martins L M, Guimaraes J E, et al. Specific downregulation of bcl-2 and xIAP by RNAi enhances the effects of chemotherapeutic agents in MCF-7 human breast cancer cells[J]. Cancer Gene Ther,2004,11(5):309-316.
[12]
Berns K, Hijmans E M, Mullenders J, et al. A large-scale RNAi screen in human cells identifies new components of the p53 pathway [J]. Nature, 2004, 428(6981):431-437.