Salivary gland adenocarcinoma(AdCa NOS) is one of the major causes of mortality among malignant salivary gland tumors. New therapeutic measure are needed to improve the outcome for patients with AdCa NOS because current therapy does not significantly improve survival rates. Transglutaminase 2(TGase 2) was implicated in forming cross-linked protein polymer, apoptosis and matrix interaction. And also TGase 2 expression is up-regulated in proliferation, migration, invasion, and metastasis of cancer cells. shRNA which has emerged as an effective method to target specific genes for silencing has provided new opportunities for cancer therapy. But there has been rarely reported using shRNA-TGase 2 transfection in AdCa NOS. The purpose of this study were to examine the specific inhibition of TGase 2 mRNA and protein expression by siRNA transfection of TGase 2 through RT-PCR and immunoslot blotting, and to study proliferation, migration and invasion assay of SGT cell line from AdCa NOS. Cell cycle analysis showed that the downregulation of shRNA-TGase 2 caused the accumulation of cells in the sub-G0/G1 phase. In migration assay, suppressing shRNA-TGase 2 inhibited the capacity of the cells to migrate compared to parental cells. In invasion assay, cells transfected with shRNA-TGase 2 decreased in invasion when compared to SGT and vector transfected cells. shRNA-TGase 2 expressing plasmids efficiently downregulated TGase 2 mRNA and TGase 2 protein expression. It suggested that the shRNA-TGase 2 targeting system against TGase 2 could have a therapeutic potentiality for malignant salivary gland tumors, especially in inhibiting and/or preventing cancer cell proliferation, migration and invasion.

Transglutaminase2 (TGM2) is a multi-functional calcium dependent enzyme that affects angiogenesis, apoptosis, differentiation, attachment, and changes in the extracellular matrix. However, its function in periodontal tissue has not yet been studied. The aim of this study was to investigate the association of the TGM2 expression and the modulation of inflammatory mediators in inflamed periodontal ligament (PDL) cells induced by pro-inflammatory cytokines such as Interleukin-1β and the Tumor necrosis factor-α. The expression of TGM2 was increased in the inflamed periodontal tissue and PDL cells. Over-expressed TGM2 in the PDL cells increased expression of MMP1, MMP3, IL-6, CXCL8, and PTGS2. Conversely, inhibition of TGM2 activity using LDN27219, a TGM2 inhibitor, resulted in decreased expression of MMP1, MMP3, IL-6, and CXCL8. The mRNA expression was confirmed by RT-PCR and quantified by qRT-PCR. Protein levels were also confirmed by immunofluoroscence staining. These results suggest that TGM2 plays an important role in the regulation of inflammatory mediators which exacerbate tissue damage in inflamed periodontal tissue.

It is not yet clear to know how normal human osteoblasts(NHost) from oral and maxillofacial area deposit, stabilize, and configure their extracellular matrix on dental biomaterial surfaces. Therefore it is necessary to design biomaterials with improved biocompatibility that will promote earlier bone differentiation and mineralization. There is now increasing evidence that TGase 2 may play a role in the initiation and regulation of the mineralization processes and serves to cross-link osteocalcin and osteopontin, which are predominant substrates for TGase 2 expressed during bone mineralization. But it is still unclear as to which TGase 2 is expressed by NHost in vitro bone formation. The purpose of this s tudy was t o determine the level of TGase 2 expression associated with collagen I , osteopontin and osteocalcin in NHost cell lines from oral and maxillofacial area in vitro. We will investigate whether TGase 2 has an active role in the biocompatibility of dental biomaterials during differentiation and mineralization of NHost. NHost cell lines were cultured under DMEM with 10% FBS at 37ﾟC and 5% CO2. Collagen quantification, mineralization and calcium assay, ALP activity assay, and RT-PCR analysis during bone differentiation and mineralization were done. ALP levels showed higher activity under AA+hGP t han under AA. I nhibition o f T Gase 2 by cystamine showed d ecreased Ca++ concentration, c ollagen I deposition and ALP level during 11 days of differentiation. TGase 2 mRNA expression of NHost was constant during mineralization stage. TGase 2 enzyme activity was increased during differentiation. Osteopontin mRNA expression during mineralization was higher than that of osteocalcin and collagen I . It suggested that TGase 2 associated with collagen I, osteocalcin and osteonectin might play a role in the differentiation of NHost from oral and maxillofacial area, but a little involved in mineralization.

Transglutaminase 2(TGase 2) expression is modulatecl by π>JF- a in various carcinoma. The role of TGase 2 ancl TNF- a expression in salivaIY gland tumors is not clear yet. Establishecl SGT cellline has been used to study the pathogenesis of salivaIY gland adenocaI‘cinoma on a cellular level in vitro. 까le pupose of this study were to examine n버NA expression of TGase 2 and TNF- ain SGT cellline comparecl to other tumor celllines, ancl to apply these results to the paùlogenesis of salivary gland tumor. After SGT, SCC-15, HN 4, and HeLa tumor celllines were culturecl under preconfluency, ancl 3 clays after postconfluency, the cells were harvested for total RNA extraction and cDNA preparation. RT-PCR for semiquantitative mRNA analysis was done. 까le obtained results were as follows. 1. TGase 2 and π>JF- amRNA expression was not induced by confluency in all the celllines 2. TGase 2 and π'JF- amRNA expression was variable but markeclly enhanced 비 SGTcellline 3. TGase 2 n버NA expression appeared to be associated with that of π>JF- ain SGT cellline From the aboving res ults, mRNA expression of TGase 2 and TNF a should play an important role in the pathogenesis of SGT cellline originated ti'om ductal cell.