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The Location and Expression in Mouse Heptocarcinoma Cell Lines with High and Low Lymphatic Metastatic Potentials.

Author: WeiYuanZuo
Tutor: TangJianWu
School: Dalian Medical University
Course: Pathology and Pathophysiology
Keywords: SNCG Heptocarcinoma Lymphatic Metastasis
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 3
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Abstract

Objective: Liver cancer is one of the most common malignant tumor in our courntry. Metastasis is a distinguished feature of its malignant nature. The molecular and pathological mechanisms are still vague now. However, the local lymphatic metastasis is a early signal in many primary tumors. In our group, we try to find out the mechanism of lymphatic metastasis through the study of mouse heptocarcinoma ascite cell lines with high and low lymphatic metastatic potentials named Hca-F and Hca-P. Previously, we found synuclein gamma (SNCG) have a higher level of mRNA and protein in Hca-F than Hca-P with gene chip and proteomics. That trend suggests that SNCG could play an importent role in metastasis. SNCG was initially found as a typical overexpression protein in breast cancer. With the further study during these years, more and more results show that SNCG affects proliferation, metastasis and drug resistance in many tumors. In this paper, we focus the location and expression of SNCG in mouse heptocarcinoma cells Hca-F, Hca-P and a mouse breast cancer cell 4T1 with a label protein named FLAG fused to SNCG gene.Methods: We tested the Hca-F and Hca-P cells with immunocytochemistry and Western Blot for location and expression of SNCG. And we tested the fused 4T1 cells with immunofluorescence and Western Blot for location, expression and polymer of SNCG.Results: The immunocytochemistry result of Hca-F and Hca-P: SNCG protein expresses in Hca-F and Hca-P cells. It locates in cytoplasma mainly and expresses in agglomerates. There is also some expression in nuclear. The strength of expression is higher in Hca-F than Hca-P. The intensity of Hca-F is +++, and that of Hca-P is ++. Randomly select 10 scopes to test different expression of SNCG, there are 91 cells out of 100 being positive in Hca-F and 74 cells out of 100 being positive in Hca-P. The data has significantly difference (p=0.000).The Western Blot result of Hca-F and Hca-P: We found a specific band at 43kD. Analyse the band with software, the expression in Hca-F is 1.9 times of that in Hca-P and the data has significantly difference (p=0.016). The immunofluoresence of 4T1 cell: The pictures show that the vector has been transfected into 4T1 cells and expressed. Cy3 gave sharp fluorescence showing SNCG locates in cytoplasma mainly. And some nuclears gave fluorescence with the DAPI suggesting the location of nuclears which means SNCG can also express a little in nuclears.The Western Blot result of 4T1 cell: We found two specific bands at 13kD and 23kD. The former is monomer of SNCG protein and the latter is dimer. Analyse the bands with software, the expression of dimer is 1.12 times than expression of monomer. The data has no significantly difference (p=0.06).Conclusion: 1. SNCG locates in cytoplasma mainly and nuclear in mouse heptocarcinoma cells and breast cancer cells.2. The expression of SNCG protein in Hca-F is 1.9 times higher than that in Hca-P.3. SNCG protein forms monomer and dimer in mouse breast cancer cell.

The Location and Expression in Mouse Heptocarcinoma Cell Lines with High and Low Lymphatic Metastatic Potentials.

Abstract

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