The Regulation Of KCa3.1on Cell Proliferation, Cell Cycle,Apoptosis And Angiogenesis Of Hepatocellular Carcinoma

Objective:To confirm the existence of KCa3.1in primary hepatocellular carcinoma, and clarify its expression features and localization.Methods:Western Blotting technology and real-time quantitative PCR assay were used to detect the expression level of KCa3.1in tissue samples. Western Blotting technology was used to detect the expression of KCa3.1in various cell lines. Immunofluorescence was used to detect the expression and localization of KCa3.1in cell.Results:The expression of KCa3.1in hepatocellular carcinoma tissue was confirmed. K comparing with para-carcinoma tissues, an increase of the transcription of KCNN4mRNA in cancer tissues was detected (P<0.05). The expression of KCa3.1was confirmed in a variety of cell lines. The KCa3.1protein was localized in the plasma membrane of hepatocellular carcinoma cell.Conclusion:The expression of KCa3.1in primary hepatocellular carcinoma was detected and its expression may be associated with the physiological function of hepatocellular carcinoma. Objective:To study the function of KCa3.1on cell proliferation, apoptosis, cell cycle and tumor characteristics of hepatocellular carcinoma. To verify the electrophysiological effects on KCa3.1of regulation chemicals.Methods:Patch-clamping technique was used to verify the blockage effects of TRAM-34on KCa3.1. MTT assay was used to research regulation function of KCa3.1on cell proliferation of hepatocellular carcinoma. Flow cytometry was used to study the function of KCa3.1on apoptosis and cell cycle process of hepatocellular carcinoma; colony forming experiment was used to study the impact of KCa3.1on colony formation capabilities of hepatocellular carcinoma. The transwell chamber assay was used to study the impact of KCa3.1on invasion and migration of hepatocellular carcinoma.Results:The blockage function of TRAM-34on KCa3.1was confirmed. TRAM-34can inhibit the proliferation, promote apoptosis and induce an arrest of Gl phase of hepatocellular carcinoma cells by blocking KCa3.1. TRAM-34can inhibit colony formation, invasiveness and migration ability of hepatocellular carcinoma cells by blocking KCa3.1. EBIO can inhibit the proliferation of hepatocellular carcinoma cells by opening KCa3.1.Conclusion:KCa3.1was involved in the regulation of liver cancer cell proliferation, apoptosis, cell cycle and malignant characteristics. The trial of channel modulators to regulate malignant process of tumor cells in vitro was successfully achieved. Objective:To study the regulation of KCa3.1on signaling pathway of hepatocellular carcinoma.Methods:Western blotting techniques were used to detect the impact effects on MAPK signaling pathway, apoptosis-related signaling pathways, cell cycle regulatory proteins and tumor cell chemokine, by the blockage of KCa3.1in hepatocellular carcinoma cells.Results:The blockage of KCa3.1can decrease the phosphorylation level of ERK1/2and increase phosphorylation level of JNK/SAPK in HepG2cells. However there is no significant relationship between KCa3.1and p38pathway. The blockage of KCa3.1can upregulate the expression of p27but not p21and cyclin E1. The blockage of KCa3.1can upregulate the activation of caspase-3, reduce the expression of bcl-2and downregulate the expression of CXCR4.Conclusion:The relationship between KCa3.1and signaling pathways on proliferation, apoptosis, cell cycle and cytokine secretion was investigated, and these results provide a theoretical basis and practical evidence for the therapeutic possibility of KCa3.1. Objective:To study the role of KCa3.1in vivo and explore its therapeutic effects.Methods:In vivo RNAi technology was used to detect effect of KCa3.1on proliferation and apoptosis of hepatocellular carcinoma. Immunohistochemistry was used to detect the impact of inhibition of KCa3.1in vivo on tumor angiogenesis.Results:The downregulation of KCa3.1induced by iRNA can reduce the proliferation and activate the apoptosis of tumor tissue. The downregulation of KCa3.1can reduce the expression of Ki67, and reduce the expression of VEGF which is related with angiogenesis.Conclusion:The verification of the inhibition on tumor formation induced by downregulation of KCa3.1was accomplished in vivo. This study provided a theoretical basis and practical evidence for the therapeutic possibility of KCa3.1.