ASC Participate In The Blood Metastasis Of Lymphoid Neoplastic P388D1 Cells

Objective: To investigate the role of ASC in P388D1 macrophage-like lymphoma neoplasm cell line by blood metastasis. It was certificated that ASC participate in the blood metastasis in P388D1 cell line and the overexpression of ASC has an inhibition effect on tumor cells after blood metastasis.Methods: (1) We firstly detected the expression of ASC in P388D1 cells by Flow Cytometry, RT-PCR and Immunofluorescence assay. (2) To extract and purify the plasmid DNA, and to overexpress ASC in P388D1 cells. For this purpose, the plasmid with ASC (pEGFP-ASC-C2) and without ASC (pEGFP-C2) were transfected into P388D1 cells, then analyzed the expression of ASC in P388D1 cells by Flow Cytometry, RT-PCR and Immunofluorescence assay. (3) The P388D1 cells with pEGFP-ASC-C2 and pEGFP-C2, and without were injected into the vena caudalis of each mouse, particularly. We observed the life span of the mice and removed lung, spleen, liver, and kidney tissue from these mice. Using HE staining method to observe the metastasis of P388D1 cells under microscope.Results:1. Both Flow Cytometry analysis and RT-PCR detection demonstrated that P388D1 cells expressed ASC. Immunofluorescence assay showed that ASC was present in the P388D1cells and located in cytoplasm.2. Flow Cytometry analysis, RT-PCR detection and Immuno- fluorescence assay showed overexpression of ASC in P388D1cells for transfecting pEGFP-ASC-C2.3. The results in vitro indicated that overexpression of ASC inhibited P388D1 cells growth after blood metastasis, life span of which was predominance higher than the control. And the tumorigenesis was earlier and the bulk of tumor was larger for the control. It is further demonstrated that the tumor cells of mice tissues from experiment group were less but those from control group were more and obvious, especially the tumor cells of lung, spleen and liver by HE staining method under microscope analyzing.Conclusion: It indicates that the overexpression of ASC inhibited P388D1 cells growth after blood metastasis. It suggests that ASC participate in the blood metastasis in P388D1 macrophage-like lymphoma neoplasm cell line.