Arsenic trioxide upregulates UL16 binding protein 1 expression in KG1a cells through ataxia telangiectasia mutated and RAD3-related kinase pathway / 中华实用儿科临床杂志

Objective:To observe the effect of arsenic trioxide (ATO) on the expression of NKG2D ligand UL16 binding protein 1(ULBP1) in acute myeloid leukemia KG1a cells, and explore the molecular mechanism for its regulation of ULBP1 expression.Methods:KG1a cells were cultured in vitro.Then, the inhibition of KG1a cell proli-feration by different concentrations of ATO was detected by cell counting kit-8(CCK8) assay, and the expression of ULBP1 mRNA and surface protein in KG1a cells were examined by real-time RT-PCR and flow cytometry, respectively.After that, the blocking effects of ataxia telangiectasia mutated and RAD3-related kinase (ATM/ATR) inhibitor caffeine on ATO-upregulated expression of ULBP1 mRNA and surface protein expressions were investigated, and the effects of ATO on the expression of CHK1 and CHK2 proteins and their phosphorylation in KG1a cells were observed by Western blot method. Results:Different concentrations (1, 2, 3, 4, 5 μmol/L) of ATO could inhibit the proliferation of KG1a cells, which was concentration dependent, and the half inhibitory (IC 50) concentration to KG1a cells was 2.7 μmol/L.The expression of ULBP1 mRNA on KG1a cells were increased when incubated with ATO at concentration 1, 2, 3, 4, 5 μmol/L, compared without ATO group, ULBP1 mRNA expression level relatively increased respectively to (1.86±0.30) times, (3.02±0.71) times, (3.16±0.75) times, (4.80±0.70) times and (3.70±0.89) times, and the differences were statistically significant (all P<0.05). Furthermore, ATO (1, 2, 3, 4 and 5 μmol/L) upregulated ULBP1 protein expression on KG1a cells compared with that in the group without caffeine, and the differences were statistically significant (all P<0.05). After caffeine pretreat KG1a cell 2 h and ATO incubate KG1a cell 24 h, ULBP1 mRNA and protein expression levels were significantly reduced.When caffeine concentration was 8 mmol/L, ULBP1 mRNA expression level relatively reduces from (9.55±0.38) times to (6.36±0.93) times compared with that in the group without caffeine, and the difference was statistically significant ( P<0.05). When caffeine concentration was 2, 4 and 8 mmol/L respectively, the expression of ULBP1 protein was reduced from that in the group without caffein treatment (3.50±0.08) times to (2.17±0.07) times, (2.02±0.06) times and (1.75±0.06) times, respectively, and the differences were statistically significant (all P<0.05). The expression of CHK1 and CHK2 proteins decreased with the increase of ATO concentration, while p-CHK1 and p-CHK2 are increased as ATO. Conclusions:ATO upregulate the expression of ULBP1 mRNA and protein in KG1a cells, and the ATM/ATR-CHK1/CHK2 pathway may be involved in it.

Bone induction by human periodontal ligament fibroblasts transfected with bone morphogenetic protein 2 gene / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To establish human periodontal ligament fibroblasts (HPDLFs) that express bone morphogenetic protein 2 and observe their osteogenesis in vivo, so as to provide fundamental data for future study on gene therapy of periodontal defect treatment.</p><p><b>METHODS</b>The phagemid expression vector for BMP-2 (pBK-B2) was transfected into HPDLFs by using Lipofect AMINE. The BMP-2 expression was determined by immunohistochemical ABC method. The BMP-2 gene transfected HPDLFs were injected into thigh muscle of nude mice under sterile condition. Specimens were collected in 21 days after injection and underwent normal procedures for fixation, decalcification, embedding, incising and staining with HE.</p><p><b>RESULTS</b>The results demonstrated that BMP-2 protein expressed in HPDLFs after BMP-2 gene transfection. The HPDLFs transfected with BMP-2 gene could induce new bone formation at the injection site.</p><p><b>CONCLUSION</b>The results indicated that BMP-2 gene was transfected successfully and expressed efficiently in HPDLFs. The HPDLFs transfected with BMP-2 gene are capable of inducing new bone formation in vivo.</p>

Overexpression of bcl-2 protects hepatoma cell line HCC-9204 from ethanol-induced apoptosis / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the effect of overexpression of bcl-2 on ethanol-induced apoptosis of primary hepatocellular carcinoma (HCC) cells.</p><p><b>METHODS</b>The retrovirus expression vector pDOR-SB containing human bcl-2 cDNA was introduced into a human HCC cell line HCC-9204 by liposome-mediated transfection. pDOR-transfected and non-transfected HCC-9204 cells were used as controls. The expression of Bcl-2 protein by transfected HCC-9204 cells was detected by the immunohistochemical method. Then the cells were cloned with the limited dilution method continually until a monoclonal cell strain whose positive rate of Bcl-2 protein was 100% detected by flow cytometry was obtained. The killing rates of cells were detected by Methabenzthiazuron assay after the treatment of 6% ethanol for 6 h. The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry.</p><p><b>RESULTS</b>Most of the pDOR-SB-transfected cells demonstrated Bcl-2 positive signals, while no signal was found in the controls. The positive rate of Bcl-2 protein detected by flow cytometry in the obtained monoclonal cell strain, which was named HCC-bcl2, was 100% after the cells had been cloned 3 times continually. The killing rate, TUNEL index and the scale of sub-G1 apoptotic peak in HCC-bcl2 cells were all significantly lower than those in the control cells.</p><p><b>CONCLUSION</b>Overexpression of Bcl-2 protein suppresses ethanol-induced apoptosis of the HCC cell line HCC-9204.</p>

The biological effects of recombinant human bone morphogenetic protein 2 on human periodontal ligament fibroblasts / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The purpose of this study is to study the biological effects of recombinant human bone morphogenetic protein 2 (rhBMP2) on human periodontal ligament fibroblasts (HPDLFs).</p><p><b>METHODS</b>Human periodontal ligament fibroblasts were primary cultured and detected the different doses of rhBMP2 on their proliferation, alkaline phosphatase (ALP) activity, osteocalcin (OC) synthesis and formation of the mineralized nodules.</p><p><b>RESULTS</b>rhBMP2 (0.25-2 mg/ml) had no remarkable effect on the proliferation of HPDLFs. The ALP activity, OC synthesis and formation of the mineralized nodules of HPDLFs were significantly stimulated by 0.5-2 mg/ml rhBMP2.</p><p><b>CONCLUSION</b>The effects of rhBMP2 on HPDLFs are dose-dependent. Not only can rhBMP2 stimulate the expression but also promote the maturation of osteoblastic phenotype of HPDLFs.</p>

Ethanol-induced apoptosis of hepatoma cell line HCC-9204 and its relationship to Bax and Bcl-2 proteins / 细胞与分子免疫学杂志

Aim To explore the ethanol-induced apoptosis effect on hepatocellular carcinoma(HCC) cells and its relationship to the expression of apoptosis associated genes, bax and bcl-2. Methods The cytotoxic effect of 20～ 100 mL/ L ethanol on HCC cell line HCC-9204 was tested by thiazolyl-blue (MTT) assay. Then apoptosis of HCC-9204 cells was induced with 60 mL/ L of ethanol for 6 h. The morphological change, DNA breakage and the change of DNA content of different cell cycles of the apoptotic cells were detected by May-Grunwald Giemsa(MGG) staining, and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and flow cytometer respectively. The changes of expression level of Bax and Bcl-2 proteins were detected by immunocytochemical staining and image analysis. Results The higher the concentration of ethanol was, the stronger the cytotoxic effect on HCC-9204 cells was. 60 mL/ L of ethanol could lead to obviously morphological apoptotic changes of HCC-9204 cells, and majority of the cells were TUNEL positive by TUNEL labeling assay. Typical apoptotic sub G1 peak was observed by flow cytometer. The level of Bax protein expression increased significantly after induced with 60 mL/ L of ethanol for 6 h, no expression of Bcl 2 were found before and after induced with ethanol. Conclusion Low dose of ethanol can induce apoptosis of HCC-9204 cells obviously, and occurance of the apoptosis is related to the increase of the level of Bax protein expression.

Expression and Significance of Ets－ 1 in Chondrosarcoma and Osteochondroma of the Jaws / 上海第二医科大学学报

ObjectiveTo analyze the expression and significance of Ets－ 1 (E26 transformation－specific) in chondrosareoma and osteochondroma of the jaws.MethodsEts－ 1 expression was de-tected by immunohistochemistry in 20 cases of chondrosarcoma and 8 cases of osteochondroma of thejaws.Results12 of the 20 cases of chondrosarcoma (t50%) showed an Ets－ 1 positive reastion. Thepositive rates of grade Ⅱ and grade Ⅲ were significantly higher than that of grade Ⅰ ( P < 0.05 ). Re-current cases showed a higher Ets － 1 positive rate than primary ones ( P < 0.05 ). Ets － 1 was presentin 12.5 % (1/8) osteochondroma. The positive rate was significantly different between chondrosarcomaand osteochondrorna ( P < 0.05 ).ConclusionThe results indicate that Ets－ 1 is overexpressed inchondrosarcoma of the jaws and correlated with its pathological grade and recurrence.

The significance of DNA content in oral squamous cell carcinoma / 实用口腔医学杂志

Objective: To study the significance of DNA content in oral squamous cell carcinoma (SCC).Methods: DNA content and cell cycle of the cells in 18 cases of oral SCC were analized by image cytometry (ICM). Results: The DNA index (DI) and proliferation index (PI) were remarkably heigher in SCC than in normal epithelium ( P