Short interfering RNA directed against the GOLPH3 gene enhances the effect of chemotherapy against oral squamous cell carcinoma by regulating Caspase3, Bcl2 and cytochrome-c expression.

Growing evidence reported that Golgi phosphoprotein 3 (GOLPH3) was involved in the progression of several human cancers. To determine whether knockout of GOLPH3 enhances the effect of Chemotherapy against cell growth of oral squamous cell carcinoma in vitro. OSCC cells were transfected with Golph3 plasmid, Golph3-RNAi and the relative control plasmids. Transfected Tca-8113 cells treated with cis-Dichlorodiamineplatinum (DDP; 0, 0.05, 0.25, 1.25, 6.25 and 31.25 ug/ml) or Paclitaxe (0, 2, 10, 50, 250 and 1250 nM) or Adriamycin (0, 0.25, 0.5, 1, 2 and 4 ug/ml) for 24 h, respectively, was determined using MTT assay. Apoptosis-related protein expression Cytochrome-C, Caspase3 and Bcl-2 was analyzed by RT-PCR and western blots. Result of MTT showed that Golph3-RNAi transfected Tca-8113 cells enhanced the effect of chemotherapy, and the effect was strengthened with the increasing concentration of drugs, and the Golph3 plasmid transfected Tca-8113 cells showed the opposite effect. RT-PCR and western blots assays revealed that expression of cytochrome-C and caspase3 were up-regulated, while Bcl-2 expression was down-regulated in Golph3-RNAi transfected Tca-8113 cells. Taken together, this study demonstrated that GOLPH3 had potent pro-tumor growth and decreased the effect of Chemotherapy, and its mechanism is primarily via cell anti-apoptosis, down-regulating the expression of cytochrome-C and caspase3, up-regulating Bcl-2 expression.

HLA-G1 and HLA-G5 isoforms have an additive effect on NK cytolysis.

The immunotolerant HLA-G could generate seven isoforms including HLA-G1--G7. The suppressive function of either HLA-G1 or HLA-G5 isoform to NK cell cytolysis has been well established. Whether HLA-G1 and HLA-G5 isoform have an additive effect on the cytolysis of NK cells remain to be explored. In this study, effects of expression of HLA-G1 and HLA-G5 isoforms and their combination on NK cytolysis was investigated. NK cell cytolysis was analyzed by detecting the NK cell surface CD107a expression. In this study, data showed that the inhibition capacity is dependent on the level of both HLA-G1 and HLA-G5 expression, but the HLA-G5 isoform has a more potent inhibition effect on the NK cytolysis (p<0.01). Furthermore, HLA-G1 and HLA-G5 have an additive effect on the suppression of NK cell cytolysis. Our study provided further understanding for the roles of HLA-G1 and HLA-G5 isoform expression in target cells immune escaping from NK cells.

[Experimental study on the reconstruction of mandibular defects with sinboneHT bone replacement in rabbits].

OBJECTIVE: After sinboneHT bone replacement (SBR) was implanted in animals, to evaluate the biocompatibility of SBR and compounded in autogenetic bone in the proportion of one to one in order to prepare for the clinical applications in the future. METHODS: Bone defects of 10 mm x l0 mm x 2 mm was made at the mandibular of rabbits, then SBR with different granule diameter and autogenetic bone was compounded in the proportion of being applied in the left defects, while autogenetic bone was implanted in the right defects and nothing was used in the right reformed defects. Animals were sacrificed at 2, 4 and 8 weeks respectively. The biologic capacity was evaluated with anatomy, X-rays studies and histology. RESULTS: SBR has better biocompatibility, which can effectively accelerate the reconstruction of bone defects and help the new bone by being compounded with autogenetic bone. It provides the appropriate scaffold or template which would allow cellular infiltration, attachment and multiplication. CONCLUSION: SBR is a kind of bone substitute material with good biocompatibility. SBR compounded with self-bone has a better regeneration function.

[Effects of hypotonic solution on the proliferation, alkaline phosphatase activity and [Ca2+]i concentration of osteoblast-like cells].

OBJECTIVE: To investigate the effects of static-stretch from the hypotonic solution on the proliferation, alkaline phosphatase activity and [Ca2+]i in the osteoblast-like cells. METHODS: Mechanical loading was introduced by swelling in the hypotonic solution. In vitro cultured MG63 were incubated under continuous swelling of 277, 240 and 163mOsm for 2h, 4h, 8h, 12h, 24h and 48h. The cell proliferation was detected by MTT assay. ALPase and [Ca2+]i were determined by modified enzyme dynamic method and OCPC respectively. RESULTS: The cell proliferation, ALP activity and [Ca2+]i increased slowly under continuous static-stretch of 277 and 240mOsm. The cell proliferation was inhibited under 163mOsm, with a sharp increase of [Ca2+]i at 8h (11.383 +/- 0.111) and an increase of ALPase activity (0.326 +/- 0.002). CONCLUSION: The static-stretch induced from the hypotonic solution has an impact on the proliferation, differentiation, ALPase and Ca2+-ATPase of the MG63. The [Ca2+]i is correlated with the ALPase.