Biochemical Differences And The Expression Of 8 Kinds Of Protein Of Two Hepatocarcinoma Cell Lines: Hca-F Cell And Hca-P Cell

Metastasis is the most lethal attribute of cancer, but its molecularmechanism is still unclear because of its complicated course. Lymph nodesare often the first organ to develop metastasis. Purpose: In order to explore the mechanism of lymph node metastasis,we examine a mouse hepatocarcinoma cell line(Hca-F) with highlymphogenous metastatic potential and its syngeneic cell line(Hca-P) withlow one with proteins corrected to metastasis. We hope to exploremechanism about lymphogenous metastasis, and find the biomarkercorrected to metastasis. Methods: 1. To study the difference of cell proliferation, the growthof Hca-F and Hca-P was evaluated. 2. Cells were collected and detected forgrowth fraction with flow cytometry. 3. Cell migration ability wasevaluated by using the chemotaxis chamber. 4. The Hca-F and Hca-P tumorcell lines were inoculated in 33 mice subcutaneously to study the differenceof tumorigenesis and metastasis between the two cell lines. 5. Theexpression of 16 kinds of proteins related with tumor metastasis in Hca-Fand Hca-P were detected by Western blotting(E-cadherin, T-cadherin,N-cadherin,β-catenin,Smad4,Ezrin,VHL,FAK,c-cam1,connexin43,NF2,Smad3,CyclinB1,Aurora,MMP-2,Plk). 6. Hca-F and Hca-P celllines were stained with antibody and analyzed by immunofluorescencemicroscopy to determine 16 proteins expression and cellular localizationpatterns. 7. To explore the difference of the anti-apoptosis ability betweenHca-F and Hca-P, the expression of proteins related with apoptosis and thealteration of cell cycle in Hca-F and Hca-P were evaluated. Results: 1. Hca-F cells growth faster than Hca-P. There wassignificant difference between Hca-F and Hca-P on cell growth (P<0.05). 2.The difference of the cell cycle between Hca-F and Hca-P was as following:The G2%, S% and G2/G1 of Hca-F are 24.0, 61.6 and 1.937 respectively.The G2%, S% and G2/G1 of Hca-P are 15.9, 49.5 and 1.864 respectively. 3.There are significant differences on cell migration ability between Hca-Fand Hca-P (P<0.05). 4. The mice bearing Hca-F cells developed 75.8%metastatic lymph node, whereas the mice bearing Hca-P exhibited 15.2%metastatic lymph node. 5. The expressions of β-catenin, E-cadherin,T-cadherin, N-cadherin and Smad4 were lower in Hca-F than in Hca-P, andthe others were no significant difference in Hca-F and Hca-P. 6. Thelocation of these proteins mentioned is no difference between Hca-F andHca-P. 7. The shearing band expressions of caspase9, caspase3, PARP werelower in Hca-F than in Hca-P. Conclusion: We can draw the conclusion that Hca-F and Hca-P is aexcellent model for tumor metastasis study. Beta-catenin, E-cadherin,T-cadherin, N-cadherin, β -catenin and Smad4 maybe play someimportant role in the differences between Hca-F and Hca-P, and themechanism need further study.