The Expressions And Significances Of CLIC1 And ECH1 In Mouse Heptocarcinoma Ascite Cell Lines With High And Low Lymphatic Metastatic Potentials

Objective: From the statistics number, we get the tendency that malignant neoplasm is the first cause inducing the death of city resident and just after cardiovascular disease at abroad. Invasion and metastasis cause malignant biological behaviour, lymphatic metastasis is one of important pathway,but the function mechanism is unclear. We study mouse hepto- carcinoma ascite lymphatic metastatic cell lines:Hca-F and Hca-P to find different expressed factor may participate in lymphatic metastasis between two cell lines.genechip and protemics have identify many protein for further study. Intracellula chloride channel 1(CLIC1) and enoy1 coenzyme hydra- tase1(ECH1) are all overexpression in high lymphatic metastatic cell linesHca-F and may play important role in tumor metastasis. CLIC1 exist in soluble and integral membrane forms. Distribute in all tissue,participating in physiological process,such as cell volume homeostasis organellar acidif- ication,regulation in cell cycle and signal transduction. Recently CLIC1 is reported overexpressed in kinds of tumor,but no study about metastasis. ECH1 locate in motochondia and peroxisome, and it is the key enzyme in the second step of beta-oxidation pathway of fatty acid metabolish. Recently different expression ECH1 is reported tumors, but no reseach about ECH1 in tumor metastasis.This article is maily to study the location and expression of CLIC1 and ECH1 in two different heptocacinoma cell lines: high metastasis cell line Hca-F and low metastasis cell line Hca-Pand establish the fundament for further study.Methods:Mouse heptocacinoma ascites high metastasis cell line Hca-F and low metastasis cell line Hca-P were seleted as the experimenat objects, immunofluorescence, immunocytochemistry and Western Blotting analysise -s were carried out to detect the localizations and expressions of CLIC1 and ECH1.Results:Immunofluorescence:CLIC1 locates in membrane and cytoplasmic in Hca-F and Hca-P cells, but whether different membrane location need more study by Immunocytochemistry. ECH1 locates in cytoplasmic both in Hca-F and Hca-P.Immunocytochemistry: CLIC1 locates in membrane and cytoplasmic in Hca-F, the intensity is +++, locates in cytoplasmic and membrane in Hca-P, the intensity is++, and more membrane location exsit in Hca-F. Random select 10 scope to test different membrane location in two cell lines, it show no significantly diversity(P>0.05). ECH1 locate in cytoplasmic both in Hca-F and Hca-P, but the intensity in Hca-F is more stronger+++ than Hca-P++. ImagePro plus analysis IOD/area of CLIC1 in Hca-F is 0.48,in Hca-P is 0.42; IOD/area of ECH1 in Hca-F is 0.324, in Hca-P is 0.285Wstern Blot: We get the ratio of relative expression of CLIC1 in Hca-F and Hca-P, then take the relative expression in Hca-P as the standard. The results show that the CLIC1 overexpressed in Hca-F and the ratio is 1.6, significantly more than in Hca-P p<0.05.ECH1 show the same result, that overexpression in Hca-F and the ratio is more than 1.5, significantly different expression in two cell lins(p<0.05)Conclusions: CLIC1 and ECH1 are both expressed in Hca-F and Hca-P cells, howver, their expressions in Hca-F are much higher, which is consistent with our previous results. Results form current works indicate that the CLIC1 and ECH1might play potential important roles in tumor metastasis.