BPOZ inhibits proliferation of breast cancer cell lines / 军事医学

Objective To investigate the effect of BTB/POZ ( broad complex, tramtrack and bric a brac/poxviruses and zinc finger) on proliferation of breast cancer cell lines.Methods The eukaryotic expression plasmid pCMV-HA-BPOZ was constructed by cloning from cDNA of human genome.Western blotting was used to detect the expression of pCMV-HA-BPOZ.Cells growth assay was used to detect the effect of BPOZ on proliferation and colony formation assay was used to detect the effect of BPOZ on cell growth ability.Results Western blotting showed that HA-BPOZ was efficiently expressed in cells.Moreover, the growth ability and proliferation of cells were significantly inhibited in BPOZ overexpressed cells compared with the control cells (both P <0.05).Conclusionp CMV-HA-BPOZ plasmid is constructed.BPOZ can restrain breast cancer cell lines MCF7 and ZR-75-1 cells from proliferating and growing.The results of our study can con-tribute to the study of functions of BPOZ in breast cancer.

Molecular mechanism of Golgi protein 73 in inflammation / 军事医学

Objective To study the effect of Golgi protein 73(GP73) on inflammation, and to reveal the effect of GP73 on tumorigenesis and metastasis.Methods The transcriptional activity of NF-κB and the expression of IL-1β, IL-6 and TNF-αwith GP73 overexpression or knockdown were detected to illuminate the role of GP73 in inflammation.According to the TCGA database, the correlation between the transcriptional activity of GP73 and the expression of NF-κB, IL-1β, IL-6 and TNF-αwas analyzed to determine the role of GP73 in tumor inflammation.Results Correlative analysis showed that there was a positive correlation between the expression of GP73 with NF-κB, IL-1β, IL-6 and TNF-α.The transcriptional activity of NF-κB was upregulated by GP73 overexpression, but downregulated by GP73 knockdown.The expression of IL-1β, IL-6 and TNF-αwas upregulated by GP73 overexpression.Ammonium pyrrolidinedithiocarbamate ( PDTC ) was in-volved in inflammation reaction induced by GP73.Conclusion GP73 is possibly involved in inflammation and promotes tu-morigenesis and metastasis.

Construction of H22 GP73 knockout gene stable strain using CRISPR/Cas9 gene editing system and identification of functions / 军事医学

Objective To knock out the GP73 gene in H22 cells originating in mice using CRISPR/Cas9 gene editing system and construct H22 GP73 gene knockout stable strain for identification of its functions .Methods Two pairs of sgRNAs that could specifically identify the upstream and downstream of GP 73 gene first promoter were designed before a recombinant eukaryotic expressional plasmid was constructed using pX 459 .After enzyme digestion and sequencing , two pairs of recombinant plasmids were co-transfected into H22 cells before puromycin was used to screen positive cells and monoclonal cells which stably knocked out GP 73 gene were developed .The knockout effect was measured by Western blotting.Cell Titer 96? AQueous One Solution Assay was used to detect the effect on cell reproductive capacity when the GP73 was knocked out .The transferability was detected through wound healing test .Results The result of Western blotting suggested that GP73 protein was undetected in the construction of H22 GP73 knockout gene stable strain after transfection.The transfer and reproduction slowed down .Conclusion H22 GP73 gene knockout stable strain can be successfully built using CRISPR/Cas9 gene editing system ,thus facilitating studies on the function of GP 73 in hepatocarcinogenesis .