| BackgroundLymphoma is a kind of lymphatic hematopoietic system malignant tumor, due tolymphocyte’s clonally hyperplasia. It mainly happens in lymph nodes, but it can also occurs inother organs, such as lung, stomach, intestines, skin, reproductive organs, bone, bone marrowand brain, etc. Lymphoma can be divided into Hodgkin’s lymphoma and non-hodgkin’slymphoma. In recent years, the incidence of malignant lymphoma shows ascendant trend, andthe B cell lymphoma takes the big part. In addition, a large number of research found that theoccurrence, development, prognosis and the biological behavior of the lymphoma is closelyrelated to the disorder of the cell apoptosis. Apoptosis is controlled by apoptosis relatedgenes.At present, we already know some malignant lymphoma’s apoptosis related genes,such as Survivin, Caspase, etc[1-3].TF-1cell apoptosis-related gene19(TFAR19) is an important new apoptosis relatedgene cloned from TF-1cells (one kind of human leukemia cell strain) by researchers in thecenter of human disease gene of Beijing university[4]. The international human gene namedcommittee named it PDCD5(programmed cell death5). PDCD5located in chromosome19q12-q13.1. The totally length of the gene is about6KB. PDCD5contains five introns andsix extrons, the full length of the cDNA is590bp. It has been experienced that PDCD5is animportant widely expressed and conservative evoluted apoptosis related gene. Not only thisprotein incrementally express in the apoptosis process, but also obvious nuclear inversionphenomenon can be seen in the early stage of the apoptosis processs. So PDCD5may be seenas a new kind of early signs of apoptosis. Because PDCD5may participate in the apoptoticprocess by reducing Survivin protein and increasing Caspase activity[5],its anomalyexpression is often closely related to tumor’s formation and development process. Accordingto reports in the literature, PDCD5’s abnormally expression exists in leukemia, liver cancer,gastric cancer, breast cancer, cervical cancer and many other kinds of tumor cells. Butwhether the level of PDCD5protein is abnormal in malignant lymphoma cell strain is stillunknown.ObjectiveThe objective is to study the expression levels of the PDCD5in the supernatant of humanlymphoma B-cell SU cells, SU/DXM cells and the SU cells induced by a variety of different concentrations of chemotherapy drugs.Research MethodsChapter I The biological characteristics of human lymphoma B-cell SU cellsObserve SU cells’ cellular morphology and the growth condition under microscope;Draw the cell growth’s curve by Cell counting method; Test lymphoma B-cell SU cells’surface markers and analysis human cell cycle distribution by Flow cytometry instrument.Chapter II The biological characteristics of SU/DXM cells.Induce SU cells by Dexamethasone in vitro, and observe SU/DXM cells’ cellularmorphology and the growth condition under microscop; Draw the cell growth’s curve by Cellcounting method; Test SU/DXM cells’ surface markers and analysis human cell cycledistribution by Flow cytometry instrument; Analysis drug-resistant spectrum and IC50valueby MTT.Chapter Ⅲ The expression of PDCD5in SU cells and SU/DXM cellsExtraction the supernatant of SU cells and SU/DXM cells.Then use PDCD5enzymelinked immunosorbent assay kit to test whether PDCD5express in the supernatant of theabove two kinds of cells.Chapter IV The expression of PDCD5in SU cells induced by several kinds of differentconcentrations of chemotherapy drugs.Use different concentrations of DXM, VP-16, L-Asp and ADM to induce SU cells whichare in logarithmic phase, and extract the supernatant. Then detect the expression of PDCD5in the supernatant by enzyme linked immunosorbent assay kit.ResultsChapter I The biological characteristics of human lymphoma B-cell SU cells1. The cell type of SU cells:The nutrient solution of SU cells is clear and the cells growsuspendedly.SU cells are round, and the cell volume is larger.2. The growth state of SU cells: To draw SU cell’s survival curve according the cellcounting method. The result of the Cell cycle detection shows the ratio as follows: G0/G1phase cells accounts for43.6%, S phase cells accounts for48.5%, G2/M phase cells takes7.97%of the cellular cycle. Different cell cycles of the SU cells present different growthconditions. SU cells express highly of B cells related surface markers, such as CD10, CD20,CD22and CD38. So SU cells belong to B cell lines.Chapter II The biological characteristics of SU/DXM cells.1. Induced by DXM in vitro, SU cells become the dexamethasone-resistant SU/DXM cells. SU/DXM cells’ morphology and growth state have no obvious difference with SU cells.SU/DXM cells are still round and grow suspendedly.And the size doesn’t change. However,several little cavity can be seen in the nucleus.2. The Cell cycle detection shows the ratio as follows: G0/G1phase cells accounts for50.9%, S phase cells accounts for35.3%, G2/M phase cells takes13.8%of the cellular cycle.In addition, SU/DXM cells also express highly of B cells related surface markers. SoSU/DXM cells still belong to B cell lines3.The drug resistance index for SU cells to DXM, Vp-16, ADM and L-ASP are8.68,1.37,3.92and3.56times than before.Chapter Ⅲ The expression of PDCD5in SU cells and SU/DXM cells.The expression level of PDCD5protein in the SU/DXM cells’ supernatant is lower thanthe SU cells’, respectively（10.67±1.48）ng/ml and（19.74±2.99）ng/ml, The differences havestatistical significance (t=8.504, P <0.05).Chapter IV The expression of PDCD5in SU cells induced by different concentrations ofseveral kinds of chemotherapy drugs.The supernatant extracted from the SU cells which are respectively induced by differentconcentrations of DXM, Vp-16, ADM and L-Asp can be detected of PDCD5proteinexpression. What’s more, the expression level is positively correlated with the concentrationof DXM and Vp-16.Conclusion1. Due to Long-term induction by DXM in vitro, SU cell can happen MDR.2. PDCD5protein expressed in the supernatant of SU/DXM cells is significantly lowerthan the SU cells’ supernatant.3. The expression of PDCD5protein can be detected in the supernatant which isextracted from the SU cells which are respectively induced by different concentrations ofDXM, Vp-16, ADM and L-Asp. And the expression level is positively correlated with theconcentration of DXM and Vp-16. |
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