ABSTRACT
<p><b>OBJECTIVE</b>To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells.</p><p><b>METHODS</b>The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry.</p><p><b>RESULTS</b>A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells.</p><p><b>CONCLUSION</b>A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.</p>
Subject(s)
Humans , 12E7 Antigen , Antigens, CD , Genetics , Base Sequence , Cell Adhesion Molecules , Genetics , Cell Line, Tumor , Cloning, Molecular , Genetic Vectors , Genetics , Hodgkin Disease , Metabolism , Pathology , Jurkat Cells , Molecular Sequence Data , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To construct a recombinant lentivirus harboring RNA interference sequence targeting mouse CD99 antigen-like 2 (mCD99L2) gene and observe its infection efficiency of 293FT cells.</p><p><b>METHODS</b>Four pairs of small interfering RNAs (siRNAs) targeting mCD99L2 cDNA were designed, synthesized and linked to the lentivirus vector SD1259 to construct the lentivirus shuttle plasmids. After sequencing, the 4 lentivirus shuttle plasmids were transfected into 293FT cells in the presence of packaging plasmids. Forty-eight hours later, the supernatant was collected and the titer and infection efficiency of the recombinant lentivirus were determined according to the expression of the reporter gene enhanced green fluorescent protein (EGFP) under fluorescent microscope.</p><p><b>RESULTS</b>DNA sequencing demonstrated that mCD99L2 siRNAs were successfully cloned to the lentiviral vector SD1259. The titer of concentrated virus was 1x10(7)/ml in the supernatant of the infected cells.</p><p><b>CONCLUSION</b>The recombinant lentivirus containing siRNA targeting mCD99L2 gene has been successfully constructed, which provide the basis for future establishment of visualized cell model and animal model of Hodgkin's lymphoma.</p>
Subject(s)
Animals , Mice , 12E7 Antigen , Antigens, CD , Genetics , Base Sequence , DNA, Complementary , Genetics , Genetic Vectors , Green Fluorescent Proteins , Genetics , Metabolism , Hodgkin Disease , Pathology , Lentivirus , Genetics , Metabolism , Molecular Sequence Data , RNA Interference , RNA, Small Interfering , Genetics , Recombinant Proteins , Genetics , Tumor Cells, CulturedABSTRACT
<p><b>OBJECTIVE</b>To evaluate the therapeutic efficacy of intensity-modulated radiation therapy(IMRT) combined with Delisheng injection for treatment of nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Sixty-six patients with pathologically confirmed NPC (stage II and III) were randomized into therapeutic group and control group. Patients in the therapeutic group were treated with Delisheng injection in addition to IMRT and those in the control group with IMRT alone.</p><p><b>RESULTS</b>No significant difference in the response rate occurred between the two groups. The incidence of adverse effects was significantly lower in the therapeutic group than in the control group, and the humoral immunity was improved in the former.</p><p><b>CONCLUSION</b>Delisheng injection can decrease the side effects of IMRT and improve humoral immunity in NPC patients.</p>